IPPCongress 2015
3rd International Plant Physiology Congress Challenges and Strategies in Plant Biology Research
December 11-14, 2015 Convention Centre, JNU New Delhi, India
1958
Indian Society for Plant Physiology Jawaharlal Nehru University National Institute of Plant Genome Research New Delhi, India Published by: Indian Society for Plant Physiology F-4, A Block, NASC Complex IARI, New Delhi-110012
Publication Committee Dr. R.K. Sairam Dr. Ajay Arora Dr. Sanjay Kumar Dr. Rita Sharma Dr. Viswanathan Chinnusamy Dr. Suresh Nair Dr. Sneh L. Singla-Pareek Dr. Pramod Kumar Dr. N. Raghuram Dr. Alok K. Sinha Dr. Rabi N. Sahoo
The views expressed in this publication by the authors are their own and these do not necessarily reflect those of the organizers
Cover page designed by Prof. Sanjay Kapoor, Department of Plant Molecular Biology, University of Delhi, South Campus, New Delhi. Printed at Cambridge Printing Works, B-85, Naraina Industrial Area, Phase-II, New Delhi-110028, Tel. 011-25891262 OUR SPONSORS
President of India Rashtrapati Bhavan New Delhi-110004
November 09, 2015
Shri Pranab Mukherjee
MESSAGE
The President of India, Shri Pranab Mukherjee, is happy to know that the Indian Society for Plant Physiology in association with Jawaharlal Nehru University and National Institute of Plant Genome Research, New Delhi is organizing the 3rd International Plant Physiology Congress on “Challenges and Strategies in Plant Biology Research” from December 11-14, 2015.
The President extends his warm greetings and felicitations to the organizers and participants and sends his best wishes for the success of the event.
Deputy Press Secretary to the President
Tel. (O): 91-11-23793528 (Direct), 91-11-23015321 Extn. 4442, Fax : 91-11-23010252 Email : [email protected] Minister of State of Agriculture and Farmers Walfare Government of India, Krishi Bhawan New Delhi-110001
November 20, 2015
Sanjeev Kumar Balyan
MESSAGE
I am immensely pleased to know that the “3rd International Plant Physiology Congress: Challenges and Strategies in Plant Biology Research” is being organized by the Indian Society for Plant Physiology in association with Jawaharlal Nehru University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi.
The theme of the International Congress is quite appropriate to the present scenario of changing climate. Fast changing global climate is posing tougher challenges to the crop scientists in terms of several abiotic stresses. In future, it is going to be the synchronized efforts of the Breeders, Crop Physiologists, Agronomists and Molecular Biologists to work on a common platform to find out different strategies to increased crop production. Understanding the basic physiological processes and the possible interventions at different steps are further to be explored at molecular level.
I am sure that the congress would provide a good opportunity to the youth to expose themselves to various research areas.
I wish the event a grand success.
( )
199-Q, Krishi Bhawan, New Delhi-110001, Tel : 91-11-23782343, Fax : 91-11-23074190 Secretary Government of India Ministry of Science and Technology Department of Science and Technology
November 18, 2015
Ashutosh Sharma
MESSAGE
I am glad to know that the “3rd International Plant Physiology Congress: Challenges and Strategies in Plant Biology Research” is being organized by the Indian Society for Plant Physiology in association with Jawaharlal Nehru University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi.
We have seen time and again as to how science can be effectively put to work to raise agricultural productivity. In India, the "Green Revolution" ushered in an era of food sufficiency and rural prosperity. The "Green Revolution" was a success due to the introduction of improved seeds, irrigation and plant protection measures combine with a visionary and positive policy support, liberal public funding for agricultural research and development, and dedicated work of farmer.
Although technological interventions can provide a new hope for Indian agriculture, there are two factors that are likely to be crucial determinants on whether or not we will be able to ultimately use this technological prowess to gain actual benefits. The first is the issue of public concerns on safety, especially with respect to health and environment. The second is the restrictive role of IPR in actually denying access of these technologies to the developing countries. I am sure that appropriate deliberations will be held among other things, on both these issues, during the Congress.
I wish the organizers of this Congress all success in meeting their objectives.
(Ashutosh Sharma)
Technology Bhavan, New Mehrauli Road, New Delhi - 110016 Tel: 91-11-26511439/26510068, Fax: 91-11-26863847, Email: [email protected], website: www.dst.gov.in Secretary and Director General Government of India Department of Agricultural Research & Education and Indian Council of Agricultural Research Ministry of Agriculture and Farmers Welfare Krishi Bhavan, New Delhi-110001
October 27, 2015
S. Ayyappan
MESSAGE
I am happy to know that the Indian Society for Plant Physiology in association with Jawaharlal Nehru University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi is organizing the “3rd International Plant Physiology Congress: Challenges and Strategies in Plant Biology Research” from December 11-14 at, JNU, New Delhi, India.
The proposed International Congress has great relevance for the 21st century because for feeding the burgeoning population of India and for attaining food security mission oriented efforts for augmenting food productivity and its sustainability through physiological, genetic and biotechnological approaches are critical. Plant Physiologists have a key role to play by providing deeper understanding of the physiological processes and their effective manipulation for improving the productivity of crops.
I hope that this International Congress will address such important issues and come out with new directions for researchers for tailoring productive plant types and that would help break the yield barrier and thereby ensure sustained food and nutrition security.
I wish the Congress all success.
(S. Ayyappan)
Tel.: 91-11-23382629; 23386711; Fax: 91-11-23384773; Email: [email protected] Deputy Director General (Crop Science) Indian Council of Agricultural Research Krishi Bhawan, Dr. Rajendra Prasad Road New Delhi-110001
November 16, 2015
Jeet Singh Sandhu
MESSAGE
It is a matter of immense pleasure to know that the “3rd International Plant Physiology Congress: Challenges and Strategies in Plant Biology Research” is being organized by the Indian Society for Plant Physiology in association with Jawaharlal Nehru University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi.
The theme of the conference “Challenges and Strategies in Plant Biology Research” is quiet appropriate and calls for immediate attention by the Plant Physiologists and other scientists across the nation. Plant Physiologists along with breeders and molecular biologists form a cohesive group that is entrusted with the responsibility of improving the productivity of our present day crop plants keeping in view of the depleting natural resources and constantly changing environment. An understanding of the mechanisms underlying the physiological processes, the effective manipulation of these processes to improve the quality and quantity of the produce and tailoring of productive eco-specific plant types is the need of the hour.
I hope this International Congress will provide a good forum to discuss some of these issues and evolve future strategies that will enable us to deal with the problem of food and nutritional security at the national level.
I wish this congress a grand success.
(J.S. Sandhu)
Ph: (Off.) 23382545 Fax : 23097003 E-mail: [email protected] Vice Chancellor Jawaharlal Nehru University New Delhi 110067 & President Indian Society for Plant Physiology
November 16, 2015
Sudhir K. Sopory
MESSAGE
It is a matter of great pleasure that Indian Society for Plant Physiology in association with Jawaharlal Nehru University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi is organizing the “3rd International Plant Physiology Congress: Challenges and Strategies in Plant Biology Research” from December 11-14, 2015 at JNU, New Delhi, India.
The congress will provide a good opportunity to the Plant Physiologists/Biologists to review the recent progress made in the various emerging areas of Plant Physiology and to focus on some of the pertinent issues concerning plant productivity in a changing scenario of climatic regime, dwindling natural resources and fast growing human population.
Plant Physiology, has and will continue to play an important role in agricultural research and development. With the technological advances in the field of crop breeding and molecular biology, there is a need for integrated approach wherein knowledge of Plant Physiology in the areas of photosynthesis, abiotic and biotic stresses tolerance, nutrient use efficiency, post harvest physiology and so on, can be applied to address the key questions pertaining to improved crop productivity in harmony with the changing environment.
I am confident that the scientific deliberations of the congress will provide the right direction for the future strategies in overcoming the challenges faced by the agricultural sector in order to provide food and nutritional security at the international level.
I wish the congress a great success.
(S.K. Sopory) Director National Institute of Plant Genome Research Aruna Asaf Ali Marg New Delhi-110067
November 16, 2015
Akhilesh K. Tyagi
MESSAGE
I am glad to know that the Indian Society for Plant Physiology in association with Jawaharlal Nehru University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi is organizing the “3rd International Plant Physiology Congress: Challenges and Strategies in Plant Biology Research” from December 11-14, 2015 at JNU, New Delhi, India.
A rapid growth in new knowledge being generated in diverse areas of Plant Biology is apparent. It is now time to consolidate the same. Thus, the theme of the Congress “Challenges and Strategies in Plant Biology Research” is quite appropriate and reflects the current approach. Plant Physiologists have played apivotal role in deciphering functional aspects of plants to help improve the productivity of our crops. This is even more relevant today in view of the fast depleting natural resources and climate change. A deeper and precise understanding of the mechanisms involved in regulation of diverse physiological processes, the effective manipulation of such processes to improve the quality and quantity of the produce and connecting the same to societal benefitsneed immediate attention. The Congress will provide an excellent platform to discuss some of these issues and plan future strategies that will help evolve sustainable food and nutritional security,not only at the national level but also at global scale.
I understand that a large number of researchers would attend this mega scientific event and feel confident that it would be a successful and rewarding experience.
My good wishes to the Congress.
(Akhilesh K. Tyagi) Director ICAR-Indian Agricultural Research Institute New Delhi-110012
29th October, 2015
Trilochan Mohapatra
MESSAGE
It is my great pleasure to learn that the Indian Society for Plant Physiology in association Jawaharlal Nehru University (JNU), New Delhi and National Institute of Plant Genome Research (NIPGR), New Delhi is organizing the “3rd International Plant Physiology Congress: Challenges and Strategies in Plant Biology Research” from December 11-14, 2015 at JNU, New Delhi, India.
Several issues that are confronting the Indian agriculture today has to find a solution through the application of Plant Physiology such as crop tolerance towards drought, heat, salinity and nutrient stresses, greater source- sink relationship, higher micro-nutrient content in the food grain, better shelf life of vegetable and fruits apart from increase in vase life of flowers. Organizing this conference at this point of time will explore new knowledge and approaches in solving some of the chronic issues. I am of the opinion that Plant Physiology, Crop Improvement and Molecular Biology are to be integrated in a manner that we are in a position to address some of the issues in a much more precise manner than what we have been doing in the past.
I wish the Congress a grand success.
(Trilochan Mohapatra) INDIAN SOCIETY FOR PLANT PHYSIOLOGY
F-4, A Block, NASC Complex, New Delhi-110012 1958 www.ispponline.org
The Indian Society for Plant Physiology (ISPP), established in 1958, is one of the oldest academic societies in India. It owes its origin to the vision and zeal of eminent plant scientists Prof. Boshi Sen, Prof. R.D. Asana, Prof. J.J. Chinoy and Prof. K.K. Nanda and the financial support of Government of India. Since then, the society has been serving the cause of research and education in the field of Plant Physiology, allied disciplines and agriculture. The society is based at National Agriculture Science Centre of Indian Council of Agricultural Research (ICAR), New Delhi. The society is a member of Global Plant Council, a coalition of plant and crops societies from across the globe. The main focus of the society is to bring plant scientists together to work synergistically towards solving the pressing problems facing humankind and to generate a critical mass to provide a strong platform for deliberations on important issues of agricultural research, education and extension and presenting views of the scientific community as policy inputs to planners and decision makers at national/ international levels. To achieve this, the society organizes and supports national and international conferences, seminars, symposia, workshops and brainstorming sessions on important issues of plant biology and agricultural sciences. It also publishes journal and proceedings for advancement of agricultural research and development. The official publication of the Society, the Indian Journal of Plant Physiology was launched in the year 1958. Since then this journal is being published regularly and is highly acclaimed for its scientific content and indexed worldwide. ISPP has instituted various awards to recognize and promote scientists for their excellent research contribution in the areas of plant sciences and agriculture. The society bestowed fellowships to >250 eminent researchers during the last fifty years and invites applications annually during the month of July/August every year for the following awards/fellowship.
Memorial lecture awards : S.K. Sinha Memorial Lecture K.K. Nanda Memorial Lecture G.V. Joshi Memorial Lecture
Recognition awards : J.J. Chinoy Gold Medal J.C. Bose Gold Medal
Young Scientist awards : R.D Asana Gold Medal (upto 40 years) R.H. Dastur Gold Medal
Best paper award : G.S. Sirohi Award (Published in IJPP)
Fellowships Fellowship of ISSP (FISSP)
CONTENT
S.No. Title Page
Scientific Program
1. ISPP Memorial Award Lectures 1-3
2. Plenary, Lead and Oral Presentations 4-107
3. Young Scientist Presentations 109-114
4. Poster Presentations 115-357
Session A : Abiotic Stress Adaptation and Management (PA001 to PA135) 115-182
Session B : Signaling and Phototransduction (PB136 to PB165) 183-198
Session C : Genomics, Epigenomics and Regulation Biology (PC166 to PC217) 199-224
Session D : Plant Metabolism and Metabolomics (PD218 to PD240) 225-236
Session E : Biotic Stress Management and Interactions (PE241 to PE287) 237-260
Session F : Plant Nutrition Physiology and Genetics (PF288 to PF322) 261-278
Session G : Plant Growth Regulators and Applications (PG323 to PG370) 279-302
Session H : Photosynthesis (PH371 to PH388) 303-311
Session I : Genomics-assisted Plant Improvement (PI389 to PI425) 312-330
Session J : Flowering and Post Harvest Physiology (PJ426 to PJ432) 331-334
Session K : Computational and Systems Biology (PK433 to PK452) 335-344
Session L : Climate Change and Agriculture (PL453 to PL462) 345-349
Session M : Reproductive Biology (PM463 to PM478) 350-357
5. Author Index 359-370
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India biotic Stres ORGANIZATION CHAIR : Sudhir K. Sopory, Vice-Chancellor, JNU CONVENORS : Ashwani Pareek, JNU & Madan Pal, IARI
CHIEF PATRONS
Ashutosh Sharma K. VijayRaghavan S. Ayyappan Secretary, DST, New Delhi Secretary, DBT, New Delhi Secretary, DARE & DG, ICAR, New Delhi
PATRONS
Asis Datta, NIPGR, New Delhi T. Mohapatra, Director, IARI, New Delhi M.S. Swaminathan, MSSRF, Chennai Anil Grover, UDSC, New Delhi J.S. Sandhu, DDG, ICAR, New Delhi A.K. Tyagi, Director, NIPGR, New Delhi J.P. Khurana, UDSC, New Delhi D.K. Sharma, Director, CSSRI, Karnal
INTERNATIONAL STEERING COMMITTEE
Hans Bohnert, University of Illinois, USA Andy Pereira, Virginia Bioinfor. Instt. USA Rakesh K. Singh, IRRI, Philippines O.P. Dhanker, UMass Amherst, USA Himadri B. Pakrasi, I-CARES, WUStL, USA Paul J. Thornalley, Univ. of Warwick, UK Govindjee, University of Illinois (UC), USA K.G. Raghothama, Purdue Agriculture, USA Neil Turner, UWA, Australia Abdelbagi Ismail, IRRI, Philippines V.R. Reddy, USDA-ARS, Baltimore, USA Elizabeth Vierling, UMass Amherst, USA Jez Joseph, WUStL, USA Julian Schroeder, UCSD, USA Jian-Kang Zhu, Purdue University, USA Autar K. Mattoo, USDA-ARS, USA Vijay Sharma, Monsanto, St. Louis, USA Tanguy Lafarge, FARCID, France K.H.M. Siddique, UWA, Australia Mohan Singh, Univ. of Melbourne, Australia K. Rajareddy, Mississippi, USA
NATIONAL ORGANIZING COMMITTEE
A.K. Tyagi, NIPGR, New Delhi P.K. Aggarwal, New Delhi K.C. Bansal, NBPBR, New Delhi K.S. Charak, DBT, New Delhi Bharat B. Chattoo, MSU Baroda M.B. Chetti, ICAR, New Delhi Renu Khanna-Chopra, IARI, New Delhi Ranjan Das, AAU, Jorhat P.S. Deshmukh, IARI, New Delhi M.C. Ghildiyal, IARI, New Delhi Ganesh Iyer, RRC, Mumbai James Jacob, RRII, Kottayam B.L. Kakralya, SKNU, Jobner M. Kar, OUAT, Bhubaneshwar P. Anand Kumar, ANGRAU, Hyderabad M. Maheshwari, CRIDA, Hyderabad A.N.L. Majumder, BI, Kolkata A.S. Nandwal, CCSHAU, Hisar K.K. Narayanan, Metahelix Bangalore A. Narayanan, Coimbatore Ajay K. Parida, MSSRF, Chennai Deepak Pental, UDSC, New Delhi A.S. Raghavendra, Hyderabad V.S. Reddy, ICGEB, New Delhi P.C. Sharma, CSSRI, Karnal N.K. Singh, IARI, New Delhi V.P. Singh, DU, Delhi Ramesh V. Sonti, CCMB, Hyderabad G.C. Srivastava, IARI, New Delhi J.P. Srivastava, BHU, Varanasi Renu Swarup, DBT, New Delhi Inyatullah Tahir, Srinagar H.S. Talwar, IIMR, Hyderabad M. Udaykumar, UAS, Bangalore K. Veluthambhi, MKU, Madurai C. Vijayalakshmi, TNAU, Coimbatore Usha Vijayraghavan, IISc, Bangalore Usha B. Zehr, MAHYCO, Jalna M.S. Sheshshayee, UAS, GKVK, Bangalore S.R. Voleti, IIRR, Hyderabad K.B. Hebbar, CPCRI, Kasaragod Nataraja N. Karaba, UAS, GKVK, Bangalore
PROGRAM COMMITTEE
A.K. Tyagi, NIPGR, New Delhi M.Z. Abdin, Jamia Hamdard, New Delhi V. Chinnusamy, IARI, New Delhi Anil Grover, UDSC, New Delhi J.P. Khurana, UDSC, New Delhi Hemant R. Kushwaha, ICGEB, New Delhi Ashwani Pareek, JNU, New Delhi T.R. Sharma, NRCPB, IARI, New Delhi P. Verma, Bioseed Res. India, Hyderabad
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India PUBLICATION COMMITTEE
R.K. Sairam, IARI, New Delhi V. Chinnusamy, IARI, New Delhi N. Raghuram, GGSIPU, New Delhi Dr. Ajay Arora, IARI, New Delhi Suresh Nair, ICGEB, New Delhi Alok K. Sinha, NIPGR, New Delhi Sanjay Kumar, IHBT, Palampur Sneh L. Singla-Pareek, ICGEB, New Delhi R.N. Sahoo, IARI, New Delhi Rita Sharma, JNU, New Delhi Pramod Kumar, IARI, New Delhi Madan Pal, IARI, New Delhi
FINANCE COMMITTEE
V.P. Singh, IARI New Delhi Madan Pal, IARI, New Delhi G. Ramarao, Tirupati V.K. Singh, CISH, Lucknow Alok K. Sinha, NIPGR, New Delhi B.C. Tripathy, JNU, New Delhi
LOGISTICS COMMITTEE
Paramjit Khurana, UDSC, New Delhi Anjali Anand, IARI, New Delhi Sanjay Kapoor, UDSC, New Delhi Sangeeta Khetarpal, IARI, New Delhi Ishwar Singh, IIMR, New Delhi Manoj K. Sharma, JNU, New Delhi Lekshmy S., IARI, New Delhi Bhupinder Singh, IARI, New Delhi Sneh L. Singla-Pareek, ICGEB, New Delhi
REGISTRATION COMMITTEE
Ashish K. Nandi, JNU, New Delhi Sanjay Kapoor, UDSC, New Delhi S. Naresh Kumar, IARI, New Delhi Hemant R. Kushwaha, ICGEB, New Delhi Girdhar Pandey, UDSC, New Delhi Sudhir Kumar, IARI, New Delhi Manoj K. Sharma, JNU, New Delhi V.P. Singh, IARI, New Delhi Puja Rai, IARI, New Delhi
TRANSPORT COMMITTEE
Ajay Arora, IARI, New Delhi Vijay Paul, IARI, New Delhi Nirala Ramchiary, JNU, New Delhi J.P. Singh, IARI, New Delhi S.D. Singh, IARI, New Delhi Praveen Verma, NIPGR, New Delhi
ACCOMMODATION COMMITTEE
Ishwar Singh, IARI, New Delhi Ajay Arora, IARI, New Delhi Ranjit R. Kumar, IARI, New Delhi Rakesh Pandey, IARI, New Delhi R.N. Sahoo, IARI, New Delhi Dharmender Singh, IARI, New Delhi I.P. Singh, IARI, New Delhi V.P. Singh, IARI, New Delhi
FOOD COMMITTEE
S. Chakraborty, JNU, New Delhi Ajay Arora, IARI, New Delhi Madan Pal, IARI, New Delhi Neera B. Sarin, JNU, New Delhi I.P. singh, IARI, New Delhi Rana P. Singh, JNU New Delhi Sneh L. Singla-Pareek, ICGEB, New Delhi J.P. Singh, IARI, New Delhi V.P. Singh, IARI, New Delhi
CULTURAL COMMITTEE
Atul K. Johri, JNU, New Delhi Ashish K. Nandi, JNU, New Delhi Renu Pandey, IARI, New Delhi Rita Sharma, JNU, New Delhi Sneh L. Singla-Pareek, ICGEB, New Delhi Shashi Meena, IARI, New Delhi Archana Kumari, IARI, New Delhi
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India ISPP Memorial Award Lectures
Professor S.K. Sinha Memorial Lecture
Physiological and molecular basis of abiotic stress tolerance in wheat
Renu Khanna-Chopra*
Stress Physiology Lab, Water Technology Centre, Indian Agricultural Research Institute, New Delhi-110012, India Renu Khanna-Chopra *Email: [email protected]
Drought and high temperature stress are major environmental factors limiting wheat productivity worldwide. Understanding physiological and genetic basis of drought tolerance is important for breeding for drought resistance in wheat to stabilize productivity in stress environments. Both drought and heat stress enhance monocarpic senescence in wheat which impacts crop productivity. Drought and heat tolerant wheat cultivars exhibited delayed senescence in stress environments. Oxidative stress is an important component of both abiotic stresses and senescence. Our studies have shown that effective antioxidant defense plays an important role in delayed senescence in stress tolerant wheat cultivars compared to susceptible genotypes. Senescence is also influenced by reproductive sink strength in wheat. Desinked wheat plants had a slower rate of senescence associated with lower ROS levels, slower nitrogen mobilization and increased antioxidant defense compared to plants with effective sink. Drought acclimation an important component of drought tolerance is also associated with co-ordinated antioxidant defense in wheat under field condition. A population of 206 recombinent inbred lines derived from drought susceptible and drought tolerant wheat cultivars WL711/C306 was phenotyped for grain yield and related traits under water deficit and irrigated conditions in seven different environments. Novel QTLs for grain yield and components were identified on chromosome 3BL which need to be further validated and fine mapped for marker development.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 1 ISPP Memorial Award Lectures
Professor G.V. Joshi Memorial Lecture
A plant physiology route to bioeconomy along the Himlayan altitudes
Sanjay Kumar*
CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061, HP, India Sanjay Kumar *Email: [email protected]
A change in altitude accompanies alteration in environmental conditions such as partial pressure of gases including
CO2, temperature, and radiation load. Accordingly, plant species possess mechanisms encompassing genes and proteins that offer adaptive advantage for the niche on one hand and also can potentially be utilized for economic activities through technological interventions on the other hand. Our strategy revolved around identifying the plant processes suitable for industrial use. We discovered that plants exhibited biochemical plasticity to compensate for low partial pressure of CO2 at high altitude through alteration of CO2 fixation pathway. The pathway was also linked to efficient nitrogen assimilation. C3 plants fixed CO2 both through phosphoenolpyruvate carboxylase (PEPCase) and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and channelized the part of fixed carbon to capture the ammonia evolved through metabolic reactions such as photorespiration. The identified mechanism was evaluated in Arabidopsis through transgenic approach, which showed growth advantage to plants at low nitrogen and improved carbon fixation. Yet another feature of plant species in the hostile environment of high altitude was modulation of metabolic processes for quick growth and development along with operation of varied protective mechanisms. Some of the species possessed protein with unusual properties as well. For example, one of the enzymes, Cu, Zn-superoxide dismutase (Cu, Zn-SOD) tolerated autoclaving in Potentilla atrosanguinea, a plant that is distributed at altitude >3500 m. Further, the enzyme catalyzed the reaction even at sub-zero temperature of (-)10°C and also beyond 50°C. The enzyme imparted tolerance to abiotic stresses through varied processes; by lignification of vascular tissues and altering the signaling pathway as discerned in transgenic Arabidopsis. The enzyme was engineered by directed mutation of targeted amino acids to enhance the thermo-stability further. Realizing enormous industrial use of the enzyme, the technology for its production and purification was transferred to an industry at premium price. While the identified adaptive mechanism have implications for agriculture industry to develop stress tolerant plants, unique proteins will find place in suitable industry for societal benefits.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 2 ISPP Memorial Award Lectures
Professor K.K. Nanda Memorial Lecture
Tomato functional genomics in reverse: Mutants as an aid to biofortify tomato fruits
Rameshwar Prasad Sharma* R.P. Sharma Repository of Tomato Genomics Resources, School of Life Sciences, University of Hyderabad, India *Email: [email protected]
During last two decades genome sequencing of several crop species has uncovered vast repertoire of gene sequences, yet the functions of many genes remains to be determined. One of best way to understand the function of a gene is to examine how the mutations alter the function of the gene and its effect on metabolic or morphological phenotype of the plant. Currently available reverse genetics tools along with genome-wide mutagenesis and gene-indexed mutant collections have revolutionized the way in which gene functions are studied in plants. In tomato we have created EMS-mutagenized populations to investigate gene function both by forward and reverse genetic approaches. EMS- mutagenized populations were used to isolate novel mutants of tomato using TILLING (Targeted Induced Local Lesions in Genome). Using the natural accessions and mutants we examined accumulation of carotenoids and folate in tomato fruits and influence of mutation/SNP on the primary metabolome of tomato fruits. The metabolite data sorting allowed us to identify the natural accessions with biochemical extremes in the carotenoids and folate levels in tomato fruits. Our results indicated that genes contributing to folate biosynthesis in plants are recalcitrant and show little polymorphism in the gene sequences. The mutations in the diverse pathways regulating light signalling, leaf architecture and ethylene biosynthesis affected the levels of carotenoids and folate in fruits. Our results indicate that the levels of folate and carotenoids in tomato fruits are regulated by a complex intertwining of the cellular metabolic pathways.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 3 Day 1: December 11, 2015
Plenary Session-I
Regulation of membrane transport by the CBL-CIPK signaling network
Sheng Luan*
Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720, USA Sheng Luan *Correspondence: [email protected]
Almost all signal transduction processes in plants involve Ca2+ that serve as a vital second messenger. Understanding how Ca2+ mediates the cellular responses triggered by myriad environmental signals is one of the most important goals for plant biologists in years to come. Recent studies uncovered a new family of Ca2+ sensors (CBLs) that target a family of protein kinases (CIPKs), establishing a novel paradigm for Ca2+ signaling in plants. The CBL calcium sensors and CIPK protein kinases are encoded by two multigene families. In Arabidopsis (with the smallest genome among flower plants), at least 10 CBLs and 26 CIPKs are identified. Each CBL interacts with a selected repertoire of CIPKs and each CIPK interacts with one or more CBLs. Some CBLs have common CIPK targets and some CIPKs share CBL regulatory proteins. Such specific and overlapping scheme in the CBL-CIPK interaction suggests both specific and redundant functions among the members of the two gene families. Genetic analysis has begun to reveal the function of individual CBLs and CIPKs. Available results indicate that CBL-CIPK interactions form an extensive network that functions in a number of signaling pathways including plant responses to abiotic stress, nutrition status, and abscisic acid (ABA). In addition, more recent work identified protein phosphatases that down-regulate the pathways activated by CBL-CIPK network and other kinases. In particular, ion channels serve as targets for such regulations in which kinases and phosphatases form specific pairs. Examples and mechanisms of ion channel regulation by CBL- CIPK network will be presented.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 4 Day 1: December 11, 2015
Plenary Session-I
Lessons from the good and the evil in plant-microbe interactions
Heribert Hirt*
King Abdullah University of Sciences and Technology, Thuwal, Saudi Arabia and Institute of Plant Sciences of Paris Saclay, France Heribert Hirt *Correspondence: [email protected]
Plants are permanently challenged by a number of adverse environmental factors including a large variety of microbes and the innate immune system revealed to play a key role in regulating plant growth and defence. Apart from studying typical plant pathogens, we are interested in extreme cases of plant-microbe interactions, such as Salmonella typhimurium, one of the major causes of food poisoning and death in humans. S. typhimurium is often found in soil samples and can infect and propagate both in animals and plants. S. typhimurium was found to apply different strategies to infect plants, some are specific to plants and others also suppress the human innate immune system. However, the ability of a variety of plants to cope with stress also depends on their association with rhizosphere microbes, but the mechanisms of the beneficial interaction are poorly understood. In the DARWIN21 project (http:// www.darwin21.net), the biodiversity of plant root microbes and the mechanisms of enhancing the tolerance of plants to heat, salt and drought stress are investigated. In short, endophytic microbes are isolated from pioneer plants in different deserts of the world. In a high throughput screening, we select bacteria for plant growth (PGP) and/or stress tolerance promotion (STP). Different biochemical and molecular approaches are used to study the mechanisms of PGP and STP activity of selected strains. Greenhouse and field trials with a number of candidates showed enhanced stress tolerance of crop plants, confirming that beneficial endophytes offer an alternative way to do sustainable agriculture in arid regions of the world.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 5 Day 1: December 11, 2015
Plenary Session-I
Functional analysis of cryptochrome 2 (OsCRY2) and a HY5 ortholog in regulating rice development
Jitendra P. Khurana* Jitendra P. Khurana Interdisciplinary Centre for Plant Genomics & Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi 110021, India *Correspondence: [email protected]
Light plays a pivotal role in regulating plant development all through its life cycle. The photomorphogenically active light is perceived by many different sensory receptors, including red/far-red sensing phytochromes, blue/UV-A sensing cryptochromes and phototropins. In recent years, our laboratory has been focusing on the functional characterization of cryptochromes from Brassica (Cry1 and CRY2) and rice (CRY2) and an ortholog of Arabidopsis HY5 from rice, which acts as a central integrator downstream to all the major sensory photoreceptors. The transcript levels of rice CRY2 gene are not affected by light but it encodes a protein that is rapidly degraded on exposure to blue light. The OsCRY2 gene when over-expressed in rice caused rapid development of seedlings and the adult plants were semi- dwarf. Even when grown in dark, the coleoptiles of OsCRY2ox young seedlings were short and displayed early leaf emergence. Unexpectedly, both the wild type and OsCRY2ox transgenics flowered at the same time. However, when OsCRY2 gene was over-expressed in Arabidopsis, it caused hyper-photo-morphogenesis, including early flowering. The HY5 ortholog in rice, which is expected to work downstream to Cry2 as well, however, did not alter flowering time, when over-expressed both in rice and Arabidopsis but it did complement hy5 mutant of Arabidopsis and also caused strong reduction in plant height in rice. These data provide evidence that sensory photoreceptors like Cry2 and its downstream signaling components like Hy5 can be exploited to alter photomorphogenic development, including control of flowering time and plant height.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 6 Day 1: December 11, 2015
Concurrent Session A-I
Genes and germplasm for climate resilient agriculture
K.C. Bansal*
ICAR-National Bureau of Plant Genetic Resources, New Delhi-110012, India K.C. Bansal *Correspondence: [email protected]
Abiotic stresses adversely affect crop productivity worldwide. To develop abiotic stress tolerant and climate resilient crop varieties, identification of key genes and germplasm lines which can serve as trait donor is critical. We focused on cloning and functional validation of candidate genes, transcriptomics to identify novel genes and gene networks, and large-scale phenotyping to identify donor germplasm lines for abiotic stress tolerance in important crops. We cloned and validated promoters (LEA1) and genes coding for transcription factors (DREB1s/CBFs, Zinc finger proteins), signalling components (ERA1) and effector genes (osmotin, LEA4, APX, genes for osmolyte synthesis). Deployment of these genes and promoters for transgenic crops development is underway in various laboratories in the country. Further, we employed NGS and computational biology approach to unravel stress responses of rice (a C3 plant with high water requirement) and sorghum (a C4 plant with low water requirement). Comparative transcriptome of drought sensitive rice cv. IR64 and drought tolerant cv. Nagina 22 and pathway analyses revealed potential role of α-linolenic acid metabolic pathway as a novel mechanism of drought tolerance of Nagina 22. Utilisation of the large untapped gene pool is the key for climate resilient agriculture. Since heat stress threatens wheat, we phenotyped the 18,653 wheat accessions for heat tolerance and identified genotypes with very high heat tolerance as donors for development of heat tolerant wheat varieties. The genes and germplasm identified in these studies will not only help unravel the molecular mechanisms but also to develop climate resilient crops with improved yield.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 7 Day 1: December 11, 2015
Concurrent Session A-I
Enhancement of nitrogen and carbon metabolism in plants through genetic manipulation of ornithine
Subhash Minocha1*, Rajtilak Majumdar1, Boubker Barchi1, Swathi Turlapati1,2 and Rakesh Minocha2 Subhash C. Minocha
1Department of Biological Sciences, University of New Hampshire, Durham, NH 03824, USA 2USDA Forest Service, NRS, Durham, NH 03824, USA *Correspondence: [email protected]
After the twenty proteinogenic amino acids, ornithine perhaps occupies the most critical position among the non- proteinogenic amino acids in plants. It is an intermediate in the inter-conversions of glutamate and arginine on one hand, and the production of proline, polyamines, GABA and several alkaloids on the other; all products of great significance to plants. In a study involving constitutive and inducible genetic engineering of ornithine catabolism in Arabidopsis thaliana and Populus nigra x maximowiczii, and analyses of substrate flux, qPCR, transcriptomics and metabolomics, we found that ornithine, whose concentration in cells is rather miniscule (vs. other amino acids), not only regulates polyamine biosynthesis but also plays an important role in regulating the flux of nitrogen from glutamate to arginine, proline and GABA. In fact, the metabolic effects of altering Glu-Orn-Arg-Pro-putrescine flux via ornithine manipulation go far beyond the biosynthesis of polyamines and the associated amino acids. We hypothesize that ornithine plays a critical role in regulating its own biosynthesis as well as the biosynthesis of glutamate (the primary nitrogen assimilation product) in the cells. Thus the overconsumption of ornithine leads to increased assimilation of both nitrogen and carbon, and potentially increased biomass production. Understanding regulation of ornithine metabolism should improve metabolic engineering of plants for stress tolerance (via manipulation of proline, GABA, and polyamines), nutritional improvement of food and feed crops (contents of important amino acids like arginine and citrulline), enhance nitrogen assimilation (reducing environmental pollution from nitrogen fertilizer), and carbon sequestration for increased biomass production for bioenergy.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 8 Day 1: December 11, 2015
Concurrent Session A-I
Toward the identification in poplar of a possible involvement of multi- step phosphorelay partners in drought perception
Lucie Bertheau3, Inès Djeghdir1, François Héricourt1, Françoise Chefdor1, Mélanie Larcher1, 1 1 1 2 Domenico Morabito , Christiane Depierreux , Daniel Auguin , Emilien Foureau , Vincent Sabine Carpin Courdavault2, Audrey Oudin2, Gaëlle Glévarec2, Franck Brignolas1 and Sabine Carpin1*
1Laboratoire de Biologie des Ligneux et des Grandes Cultures, Université d’Orléans, UPRES EA 1207, rue de Chartres, BP 6759, 45067 Orléans Cedex 02, France Arbres et Réponses aux Contraintes Hydriques et Environnementales (ARCHE), INRA, USC1318, 45067 Orléans, France 2Laboratoire Biomolécules et Biotechnologies Végétales (BBV), Université François Rabelais de Tours, EA 2106, 31 avenue Monge, 37200 Tours, France 3Laboratoire Processus de Transferts et d’Echanges dans l’Environnement (PROTEE), Université de Toulon, EA 3819, 83957 La Garde Cedex, France *Correspondence: [email protected]
The molecular basis of plant drought perception and the resulting cellular osmotic stress are still unknown. In yeast, the HOG pathway controlled by the multi-step phosphorelay system (MSP) SLN1-YPD1-SSK1, is the best characterized osmosensing pathway, leading to yeast adaptation on high-osmolarity and high-salt media due to glycerol accumulation. In Populus, we identified MSP proteins such as a Histidine-aspartate Kinase, HK1, homologous to SLN1, Histidine- containing Phosphotransfer proteins, HPt1 to HPt10, and 9 Response Regulators (RR). By combining several approaches including yeast two-hybrid and bimolecular fluorescence complementation assays and co-expression analysis of mRNAs encoding studied proteins, we showed the existence of a partner network linked to HK1, the putative poplar osmosensor. This network involves HK1, HPt2, 7 and 9 and six transcription factors belonging to the RR-B family, RR12, 13, 14, 16, 18 and 19. Even if the “osmosensor” function of HK proteins such as AHK1 in Arabidopsis is still not clearly attested, a role in dehydration avoidance is well established. Thus, in poplar, the HK1 MSP could be involved in dehydration-sensing mechanisms and plant water status control. To go further in this study and better understand the role of this HK1 MSP in drought tolerance, we are now investigating the HK1 function by a loss of function approach based on the RNAi strategy.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 9 Day 1: December 11, 2015
Concurrent Session A-I
Abiotic stress regulates expression of Galctinol synthase genes in rice through intron retention and nonsense-mediated decay (NMD) of mRNA
Sritama Mukherjee, Sonali Sengupta, Abhishek Mukherjee and Arun Lahiri Majumder* Arun Lahiri Majumder
Division of Plant Biology, Bose Institute (Centenary Building), Kolkata-700054, India *Correspondence: [email protected]
Galactinol synthase (GolS), the first committed enzyme in Raffinose Family Oligosaccharide (RFO) synthesis pathway, produces galactinol from UDP-galactose and inositol. The two isoforms of Oryza sativa GolS (OsGolS) gene are located in chromosomes 3(OsGolS1) and 7(OsGolS2). The genes are interspersed by conserved introns with each intron harboring a premature termination codon (PTC). We identified a non-canonical regulation mechanism controlling the splicing and maturation of both the OsGolS isoforms in photosynthetic tissue. Splicing and maturation of both OsGolS transcripts are linked to abiotic stress conditions. During abiotic stress, the premature and mature transcripts of both isoforms accumulate in a rhythmic manner for very small time windows interrupted by phases of complete absence of either. Reporter gene assay using GolS promoters under different abiotic stresses does not reflect this accumulation profile, suggesting that this regulation occurs post-transcriptionally. We suggest that there is a surveillance mechanism triggering the degradation of the premature transcript preventing its accumulation in the cell. This mechanism fits the paradigm of PTC-induced nonsense-mediated decay (NMD), a less known event in plants. The detection and degradation of prematurely terminating transcripts coupled to repressed splicing can be described as a form of regulated unproductive splicing and translation (RUST). This regulatory circuit may have contributed to the adaptation of plants to stress and phloem loading as suggested by the biological role of galactinol in green tissues.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 10 Day 1: December 11, 2015
Concurrent Session A-I
From ‘Meta-OMICS’ to mutation breeding: Improving salinity and drought tolerance in rice
Rajeev K. Bahuguna1, Ray Singh Rathore1, Priyanka Das1, Sneh L. Singla Pareek2 and Ashwani Pareek1* Ashwani Pareek
1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Road, New Delhi-110067, India *Correspondence: [email protected]
Rice is a staple food crop for more then half of the world population. An estimated demand for rice would increase at least by 50 percent in the next 25 years to feed the growing population. Abiotic stresses such as salinity, drought and heat are major constraints for global rice production in the current and more so in the future climate. Taking clues from naturally salinity tolerant rice genotypes, we are trying to understand the molecular basis of stress tolerance using an OMICS-based approach. This work has lead to the identification and functional validation of key stress responsive genes, which may prove to be a suitable candidate for translational research. Taking an alternative approach of mutagenesis, we have generated several mutant lines through gamma-irradiation and screened them at seedling stage for different abiotic stress tolerance. Three screened mutant lines showed significant potential to survive under severe heat, drought and salinity stress at seedling stage. These mutant lines were further characterized for their mechanism of stress tolerance at reproductive and grain filling stage. Plants were grown in 15 L pots in the net house and phenotyping for heat, drought and salinity stress was done across the growth stages. Besides agronomic traits, key physiological traits such as photosynthesis, physiological water use efficiency, stomatal conductance, spikelet fertility, pollen germination, K+/Na+ ratio, spikelet fertility and source-sink regulation at grain filling were studied. These mutant lines could be used for improving rice production and varietal improvement for future climate.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 11 Day 1: December 11, 2015
Concurrent Session A-I
Altered Cu homeostasis regulates drought response in rice
Saurabh Raghuvanshi1*, Sonia Chhillar Balyan1, Mukesh Kumar1, Roseeta Devi Mutum1, Utkarsh Raghuvanshi1 and Saloni Mathur2 Saurabh Raghuvanshi 1Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, New Delhi-110021, India 2National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India *Correspondence: [email protected]
It is important to understand the molecular basis of natural drought tolerance that evolved in tolerant rice cultivars/ varieties. We conducted in-depth comparative analysis of drought tolerant and sensitive rice cultivars to explore the molecular basis of drought tolerance. Our study discovered a multi-tiered drought-induced network, involving transporters, TFs, miRNAs, Cu-proteins and metabolic enzymes, in the leaves that leads to enhanced ROS accumulation and stomatal closure in the tolerant cultivar. The hallmark was the down-regulation of Cu transporters in the tolerant rice cultivars (but not in sensitive one) during drought, leading to fall in internal copper levels. This kick-starts the ‘copper economy mode’ via OsSPL9 wherein a group of miRNAs (osa-miR159f, osa-miR1871, osa-miR398b, osa- miR408-3p, osa-miR2878-5p, osa-miR528-5p and osa-miR397a), referred to as DTA (Drought Trait Associated) miRNAs, are up-regulated. These DTA miRNAs target several genes coding for copper proteins such as plantacyanins, laccases and Cu-Zn SODs. Interestingly, even though internal Cu levels fall, the level of plastocyanin remains stable indicating re-mobilization of internal Cu to stabilize plastocyanin and the photosynthetic electron transport. On the other hand, down-regulation of Cu-Zn SODs in the flag leaves of the tolerant cultivars raises ROS levels ultimately leading to enhanced stomatal closure. Physiological advantage of high DTA miRNA levels was demonstrated by overexpressing MIR408 in the drought sensitive rice cultivar leading to significant enhancement in vegetative growth and tolerance to water deficit conditions. In summary, differential regulation of copper transporters during drought defines atleast one way wherein tolerant rice cultivar modulates its metabolism to better endure drought.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 12 Day 1: December 11, 2015
Concurrent Session B-I
SHW1 Interacts with HY5 and COP1, and promotes COP1-mediated degradation of HY5 during Arabidopsis seedling development
Anjil Kumar Srivastava, Dhirodatta Senapati and Sudip Chattopadhyay* Sudip Chattopadhyay Department of Biotechnology, National Institute of Technology, Durgapur-713209, India *Correspondence: [email protected]
Light acts as one of the most influential environmental factors for plant growth and development. Arabidopsis seedlings follow two distinct developmental patterns in the presence and absence of light. The dark grown seedlings have long hypocotyl with apical hooks, and with small and closed cotyledons. The light grown seedlings have short hypocotyl with open and expanded cotyledons. Downstream to photoreceptors, multiple positive and negative regulators have been identified that are involved in Arabidopsis seedling development. SHW1 works as a negative regulator of photomorphogenesis. SHW1 and COP1, an ubiquitin ligase, genetically interact in an additive manner to suppress photomorphogenesis. HY5 is a bZIP transcription factor that works as a positive regulator of photomorphogenesis at various wavelengths of light, and is degraded by COP1 in dark. SHW1 genetically and physically interacts with both COP1 and HY5. The genetic studies reveal that SHW1 and HY5 work in an antagonistic manner to regulate photomorphogenic growth. Additional mutation of SHW1 in hy5 mutant background is able to suppress the gravitropic root growth defect of hy5 mutants. Moreover, SHW1 has been shown to promote COP1-mediated degradation of HY5 through enhanced ubiquitylation in the darkness. The interplay of these light-signaling pathway regulators and their concerted role in Arabidopsis seedling development will be discussed.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 13 Day 1: December 11, 2015
Concurrent Session B-I
Light signalling and MAP kinase cascade: A crosstalk
Alok Krishna Sinha*
National Institute of Plant Genome Research, New Delhi-110067, India Alok K. Sinha *Correspondence: [email protected]
Mitogen activated protein kinase (MAPK/MPK) cascade are universally conserved cascade among eukaryotes and are known to regulate several stress responses and developmental processes. They usually consisit of three tier components, MAPK, MAPK kinas (MAPKK/MAP2K/MKK) and MAPKK kinase (MAPKKK/MAP3K/MKKK) connected to each other with an event of phosphorylation. Light signal transduction pathways have extensively been studied in plants and are known to control several developmental processes. However, a direct connection of MAPK and light signaling pathways was not reported. Here, we present a MAPK module, consisting of MKK3-MPK6 that is activated by blue light in a MYC2 dependent manner. MYC2 is a transcription factor that negatively regulates blue light signaling during Arabidopsis seedling development. MPK6 physically interacts and phosphorylates MYC2, and is phosphorylated by MKK3. Furthermore, MYC2 binds to MPK6 promoter and regulates its expression in a feedback regulatory mechanism in blue light signaling. The mutational and physiological studies illustrate the function of MKK3-MPK6-MYC2 module in Arabidopsis seedling development and provide a new mechanistic view of photomorphogenesis.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 14 Day 1: December 11, 2015
Concurrent Session B-I
CBL-interacting protein kinase, CIPK21, regulates osmotic and salt stress responses in Arabidopsis
Girdhar K. Pandey1*, Poonam Kanwar1, Amarjeet Singh1, Leonie Steinhorst5, Amita Pandey1, 1 1 1 2 3 Akhlilesh K. Yadav , Indu Tokas , Sibaji Sanyal , Boem-Gi Kim , Sung-Chul Lee , Girdhar K. Pandey Yong-Hwa Cheong4, Jörg Kudla5 and Sheng Luan6
1Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi, India 2Department of Molecular physiology and Biochemistry, National Institute of Agricultural Biotechnology, Suwon, Korea 3Deparment of Life Science, HeukSeok-Dong, Dongjak-Gu, Seoul, Korea 4Department of Bio-Environmental Science, Sunchon National Univ., Suncheon, Jeonnam, Korea 5Molekulargenetik und Zellbiologie der Pflanzen Institut für Biologie und Biotechnologie der Pflanzen Universität Münster Schlossplatz 4 48149, Münster, Germany 6Department of Plant and Microbial Biology, Univ. of California Berkeley, Berkeley, California, USA *Correspondence: [email protected]
The role of calcium mediated signaling has been intensively studied in abiotic stress responses in plants. Calcineurin B-like proteins (CBLs) and CBL-interacting protein kinases (CIPKs) constitute a complex signaling network in diverse stress responses. Osmotic stress imposed by soil salinity and drought is a major abiotic stress that impedes plant growth and development and has been shown to involve calcium-signaling processes. In this study, we report the functional analysis of the Arabidopsis kinase CIPK21 that is ubiquitously expressed in plant tissues and upregulated by multiple abiotic stress conditions. A loss-of-function mutant of CIPK21 revealed to be hypersensitive to grow on salt and osmotic stress conditions. The calcium sensors, CBL2 and CBL3, were found to physically interact with CIPK21 and target this kinase to the vacuolar membrane. Moreover, preferential localization of CIPK21 to the vacuolar membrane was detected under salt stress conditions when co-expressed with CBL2/CBL3. These finding suggests that CIPK21 regulate responses to salt stress condition inli Arabidopsis.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 15 Day 1: December 11, 2015
Concurrent Session B-I
Engineering ABA receptors to enhance WUE and drought tolerance of rice
V.V. Santosh Kumar, Shashank K. Yadav, Sanya Shrivastava, Rakesh K. Verma, Vishakha Sharma and Viswanathan Chinnusamy* Viswanathan Chinnusamy
Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India *Correspondence: [email protected]
Abscisic Acid (ABA) plays crucial roles in development and stress responses of plants. The START domain proteins PYRABACTIN RESISTANCE1 (PYR1)/PYR1-like (PYL)/Regulatory Components of ABA Receptors (RCARs) have been identified as bona fide ABA receptors (ABARs) from Arabidopsis. Towards understanding structural and functional diversity of ABAR family members from rice, we identified 12 PYR/PYL homolog ABARs from rice. Homology modeling of ABARs cloned from Nagina 22 revealed that 10 of them may function as receptors in core-ABA signalling. Docking studies were carried out with ABA, ABA metabolite, agonists and antagonists illustrated differential affinity of different ABARs to these legends. Expression analysis of ABARs revealed differential tissue specific and stress responses, suggesting their non-redundant role in stress response and development. Transgenic rice lines with constitutive/ stress responsive over expression and RNAi silencing of ABARs are being developed. Rice transgenics expressing AtRD29A::OsABAR6 showed hypersensitivity to ABA in germination as compared with non-transformed (NT) wild type and RNAi lines. This suggests that ABAR6 conferred enhanced ABA perception and thus function as ABA receptor. The RD29A:ABAR6 transgenic rice lines are more drought tolerant than NT plants in terms of maintenance of RWC, membrane stability index, chlorophyll content, survival and recovery growth in pot culture under greenhouse conditions. Water utilization experiment revealed that transgenic plants used about 40% less water per a day as compared to wild type plants. Results from this study suggest that ABARs have potential to improve WUE and drought tolerance of rice.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 16 Day 1: December 11, 2015
Concurrent Session B-I
Tetraspanin family of proteins: New regulators of development and abiotic stress response in rice
Surekha Katiyar-Agarwal1*, M. Balaji1 and Manu Agarwal2 Surekha Katiyar-Agarwal 1Department of Plant Molecular Biology, University of Delhi, South Campus, New Delhi-110021, India 2Department of Botany, University of Delhi, Delhi-110007, India *Correspondence: [email protected], [email protected]
Tetraspanins belong to a super family of highly conserved membrane proteins found in all multicellular and few unicellular organisms. Tetraspanin proteins (TETs) are comprised of four transmembrane domains, two extracellular loops with conserved cysteine residues, and two cytoplasmic tails. The dynamic nature of TETs to interact with multiple partners and form ‘tetraspanin-enriched micro domains’, and their preferential targeting to membranes emphasizes their participation in signal recognition/perception, and coordination of intracellular signaling cascades in cells. In animals, tetraspanins play crucial role in multitude of cellular processes, including cell proliferation, adhesion, fusion, movement, interaction and intracellular trafficking. Unlike animals, the biological function of TETs in plants has not been well explored. In Arabidopsis, tetraspanins are known to be associated with development processes such as leaf morphogenesis, root and floral organ formation. The present study was designed to gather comprehensive information on 15 rice tetraspanin members (OsTETs) with respect to their genomic organization, chromosomal distribution, phylogeny and domain structure, and expression pattern. Detailed analyses of OsTETs revealed that these proteins consist of signature ‘GCCK/R’ motif as found in Arabidopsis. Expression profiling studies advocated their role in rice development, including flag leaf senescence and defining plants response to several abiotic stress and nutrient deprivation conditions. Subcellular localization of eight OsTETs suggested that they reside on plasma membrane. Transgenic rice plants for altered expression of OsTET5 have been generated and their preliminary phenotypic analyses supported role of OsTET5 in abiotic stress tolerance. Detailed characterization of transgenic rice lines is being carried out and will be presented.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 17 Day 1: December 11, 2015
Concurrent Session B-I
Phosphorylation-dependent regulation of G-protein cycle during nodule formation in soybean
Swarup Roy Choudhury and Sona Pandey* Sona Pandey Donald Danforth Plant Science Center, 975 N. Warson Road, St. Louis, MO 63132, USA *Correspondence: [email protected]
Signaling pathways mediated by heterotrimeric G-protein complex comprising Ga, Gb and Gg subunits and their regulatory RGS (Regulator of G-protein Signaling) protein are conserved in all eukaryotes. We demonstrate that the specific subunits of Gá, Gb and Gg proteins of soybean heterotrimeric G-protein complex regulate signaling during nodulation. We also show the role of nod factor receptor 1 (NFR1)-mediated phosphorylation in regulation of G- protein cycle during nodulation in soybean. Our data establish that during nodulation the G-protein cycle is regulated by the activity of RGS proteins. Lower or higher expression of RGS proteins results in fewer or more nodules, respectively. NFR1 interacts with RGS proteins and phosphorylates them. Analysis of phosphorylated RGS protein identifies specific amino acids which when phosphorylated; result in significantly higher GTPase accelerating activity. These data point to a phosphorylation-based regulation of G-protein signaling during nodule development. We propose that active NFR1 receptors phosphorylate and activate RGS proteins, which help maintain the Ga proteins in their inactive, trimeric conformation, resulting in successful nodule development. Alternatively, RGS proteins might also have a direct role in regulating nodulation because overexpression of their phosphomimic version leads to partial restoration of nodule formation in nod49 mutant.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 18 Day 1: December 11, 2015
Concurrent Session C
Functional genomics of thermal tolerance in wheat
Paramjit Khurana*
Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India Paramjit Khurana *Correspondence: [email protected]
Climate change and abiotic stress affects agriculture and crop production adversely. Wheat is a temperate crop and prone to various abiotic stresses. Tolerance to heat stress is a complex phenomenon and controlled by multiple genes. Heat tolerance related gene transcripts were identified based on their putative functions and validated by cDNA macroarray and northern/RT-PCR analysis in wheat. A common response to high temperature stress is the synthesis of Heat Shock Proteins (HSPs). HSP encoding genes are found to be under the control of Heat Shock Factors (HSF), transcription factors that regulate the expression of HSP genes. One such gene, a chloroplastic small heat shock protein (sHSP26) was found essential for tolerance to heat stress. Transgenic Arabidopsis plants over- expressing wheat HSP26 were more tolerant to high temperature induced damage to the photosynthetic machinery, and the antisense transgenic plants were found to be severely sensitive to heat stress. Similarly, a new heat shock factor (HSF) possesses the characteristic domains of class A type plant HSFs was identified. Functional genomics approaches indicated plants overexpressing TaHsfA2d possess higher tolerance towards high temperatures and a higher yield and biomass under constant heat stress conditions. Analysis of putative target genes showed higher and constitutive expression of several abiotic stress responsive genes in transgenic Arabidopsis plants over-expressing TaHsfA2d. TaHsfA2d can also functionally compliment the T-DNA insertion mutants of AtHsfA2. These observations suggest that discovery of novel genes may be useful in molecular breeding of crop plants, especially wheat, to improve yield under abiotic stress conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 19 Day 1: December 11, 2015
Concurrent Session C
Genomics-assisted mapping of QTLs and genes for salt tolerance in Indian rice (Oryza sativa L.) germplasm
Nagendra K. Singh1*, Sushma Tiwari1, Sefali Mishra1, Balwant Singh1, S.V. Amitha Mithra1, 1 2 2 3 Vandna Rai , Ashok K. Singh , Vinod Kumar and S.L. Krishnamurthy Nagendra K. Singh
1National Research Centre on Plant Biotechnology, Pusa Campus, New Delhi-110012, India 2Central Soil Salinity Research Institute, Karnal–132001, India 3Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India *Correspondence: [email protected]
Soil salinity is a major constraint to rice production in large inland and coastal areas of rice cultivation around the world. Modern high yielding rice varieties are particularly sensitive to high salt stress. There are salt tolerant landraces and traditional varieties of rice but with limited information on QTLs and genes for their salt tolerance. Salt tolerance is generally quantitatively inherited because of its control by multiple genes located in different genomic regions. With the advent of DNA markers it is possible to genetically map these quantitative trait loci (QTLs) using different types of mapping populations. However, this requires genotyping of entire population using large number of polymorphic markers to create a dense molecular genetic map. Bulked segregant analysis (BSA) allows rapid identification of markers linked to the trait, but it has been applied mainly to map simply inherited traits. Here we describe a method for rapid identification of genomic regions (QTLs) for seedling and reproductive stage salt tolerance in rice using two different mapping populations of recombinant inbred lines (RILs) using a bulked segregant analysis (BSA) approach. We found that the number of RILs with extreme phenotype to be pooled for BSA was thirty each to get the maximum heterogeneity in the non-associated regions of the genome. The parents and bulks were genotyped using a specially designed 50K SNP chip to identify genomic regions showing homogeneity of alleles for polymorphic SNPs in the bulk(s). The method was applied to ‘CSR11/MI48’ RILs segregating for reproductive stage salt tolerance. Genotyping of the parents and RIL bulks, made on the basis of salt sensitivity index for grain yield, revealed 6,068 polymorphic SNPs of which 22 genomic regions (QTLs) showed contrasting allelic homogeneity in the two bulks. The method was validated in another RIL population of ‘CSR27/MI48’ used earlier for mapping QTLs for salt tolerance using low- density SSR markers. BSA using 50K SNP chip in the CSR27 populations revealed 5,035 polymorphic SNPs and 34 QTLs showing allelic homogeneity in the two bulks. The BSA not only confirmed the location of QTLs mapped earlier in this population but also identified additional QTLs because of high density SNP genotyping. BSA with high-density SNP chip provides a rapid approach to scan the whole genome for mapping QTLs for complex agronomic traits in rice. We have also identified novel sources of salt tolerance in Indian wild rice germplasm collected from different eco- geographical regions of India.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 20 Day 1: December 11, 2015
Concurrent Session C
Sequence polymorphism in the Waxy locus and its relationship with apparent amylose content of endosperm starch in cultivars of rice (Oryza sativa L.) from Northeast India
Nikhil K. Chrungoo* and Ayam Gangarani Devi Nikhil K. Chrungoo
Molecular Plant Physiology Laboratory, Department of Botany, North Eastern Hill University, Shillong, Meghalaya, India *Correspondence: [email protected]; [email protected]
The Northeast India, being at the confluence of Indo-Malayan and Indo-Chinese biogeographic realms, represents one of the world “biodiversity hotspots”. With a rich diversity of cultivars, the rice gene pool of the region shows marked variations in various quality traits including aroma and starch quality. Since starch quality is primarily determined by its apparent amylose content and that the Waxy locus controls much of the variation in apparent amylose content, the linkage between sequence polymorphism in the Waxy locus and the apparent amylose content assumes significance. Our results reveal that based on combination of “G/T” polymorphism in 5’leader intron splice site, length polymorphism in exon 2 and “A/C” polymorphism in exon 6, the Waxy locus of rice can have four allelic forms viz.
“Int1T-Ex2(23bp)2-Ex6C”, “Int1T-Ex2(23bp)1-Ex6A”, “Int1G-Ex2(23bp)1-Ex6C” and “Int1G-Ex2(23bp)1-Ex6A” which correlate specifically with “waxy”, “very low”, “low/intermediate” and “high amylo-” cultivars respectively. Phylogenetic tree on alignment matrix of nucleotide sequences of the p-SINEI element of the Waxy locus clustered the upland and japonica cultivars separately from the lowland and indica cultivars thereby establishing the role of transposons in the Waxy locus in differentiation of upland/japonica and lowland/indica cultivars.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 21 Day 1: December 11, 2015
Concurrent Session C
Orthoulogues of blast resistance gene Pi54 interacts at different domains of Avr-Pi54 gene of Magnaporthe oryzae
T.R. Sharma*, B.N. Devanna and Soham Rey T.R. Sharma National Research Center on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi-110012, India *Correspondence: [email protected]
Decoding of whole genome sequence of Rice (Oryza sativa) and its one of the important pathogens Magnaporthe oryzae have provided an excellent opportunity to understand molecular basis of host –pathogen interaction. We identified, cloned and characterized a major gene (Pi54) for blast resistance from rice using map based cloning approach. Its orthologue Pi54 of was also cloned from wild species of rice Oryza officinalis and functionally validated in different genetic backgrounds of rice. Functional complementation of the gene has confirmed its stable and high- level of resistance expression against the geographically diverse strains of M. oryzae. To understand the molecular basis of interaction between Pi54 and Avr –Pi54 genes we sequenced the whole genome of an Avirulent strain of M. oryzae and cloned Avr-Pi54. The docking experiments involving four Pi54 R proteins and an AVR-Pi54 protein revealed that all the resistance proteins could interact with AVR counterpart, whereas the susceptible Pi54tp protein does not show any interaction with AVR-Pi54. This finding is not unexpected as there is a large deletion at NBS region of Pi54tp protein. The protein molecular docking results of this study also revealed that the Pi54 of protein interacted with AVR-Pi54 only through its NBS domain. Whereas the other Pi54 protein interacted through the CC-NBS-LRR regions and Pi54rh interacted through the NBS-LRR regions. This differential preference of AVR protein for binding domains among the Pi54 proteins might be attributed to its different resistance mechanisms against M. oryzae. Among the resistant Pi54 proteins, a stress inducible protein binding domain, STI1 is present only in Pi54 of and absent in other two resistant proteins. Different results obtained on cloning and characterization of Pi54 orthologues will be discussed in this presentation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 22 Day 1: December 11, 2015
Concurrent Session C
A multigene cluster underpins a large effect QTL for rice yield under drought
Ajay Kohli* Ajay Kohli Plant Molecular Biology Laboratory, Genetics and Biotechnology Division, International Rice Research Institute, DAPO 7777, Metro Manila, Philippines *Correspondence: [email protected]
Limited QTLs have been cloned for the underlying cardinal genes. For drought tolerance in plants two QTLs have been dissected for the cardinal gene. As opposed to such single gene QTLs, which are mostly limited to minor effects on the traits, large effect QTLs have now been cloned for rice yield under drought. Our efforts at the dissection of one such large-effect QTL has revealed a gene cluster that is essential for the functionality of QTL qDTY12.1. The multigene cluster is unique in its functionality in a manner analogous to the regulon-like genetic architecture. A transcription factor and its co-localized target genes work in concert to affect the trait of drought tolerance. Along with this novel aspect, it is also the first time that a QTL from a susceptible variety has been successfully dissected at the molecular level. These results suggest that complex traits may be best addressed through complex loci rather than a single master switch.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 23 Day 1: December 11, 2015
Concurrent Session C
Next generation genomics: Connecting genes and (epi)genomic variations to abiotic stress response/adaptation
Mukesh Jain* Mukesh Jain National Institute of Plant Genome Research, New Delhi-110067, India *Correspondence: [email protected]
The availability of high-throughput sequencing technologies provide opportunity to unveil the genetic/epigenetic basis of abiotic stress response and adaptation. Rice is an important crop accounting for food security of over half the world population. Water-deficit and salinity are the major abiotic factors that affect rice productivity worldwide. Rice germplasm exhibit variability in their response to abiotic stresses, which may be attributed to genetic and epigenetic variations. Although several stress-responsive genes have been identified and a few regulatory networks have been proposed, tolerance/response mechanisms still remain unclear to a larger extent. We performed whole-genome, transcriptome and bisulphite sequencing of three rice cultivars with contrasting responses to drought and salinity stress. We identified thousands of high-quality SNPs and InDels among these cultivars. One-fourth of the SNPs and InDels were detected in genic regions and about 3.5% of the total SNPs resulted in non-synonymous changes. DNA polymorphisms present in the differentially expressed genes within the known quantitative trait loci were also identified. RNA-seq analysis identified several novel stress-responsive transcripts/isoforms. We discovered extensive DNA methylation at single- base resolution in rice cultivars, identified the sequence context and extent of methylation at each site. Numerous differentially methylated regions (DMRs) among different cultivars were identified and many of them were found associated with differential gene expression. The transposon-associated DMRs were found coupled to the transcript abundance of the transposon and/or proximal protein-coding gene(s). These results provide insights into the regulatory mechanisms underlying abiotic stress response/adaptation and functional markers/ target genes for stress tolerance.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 24 Day 1: December 11, 2015
Concurrent Session A-II
Elucidation of the molecular machinery components associated with adaptation of rice plants to heat stress
Anil Grover* Anil Grover Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi-110021, India *Correspondence: [email protected]
Previous work from our laboratory has shown that the transcriptional profile of rice (Oryza sativa L.) is significantly modulated within 10 min of heat stress (HS_10): analysis of differentially expressed genes (DEGs) revealed that 1,556 genes were rapidly up-regulated (>twofold, P<0.05) at HS_10 as compared to 600 down-regulated genes. The enrichment of GO terms protein kinase activity/protein serine kinase activity, response to heat and reactive oxygen species in up-regulated genes after HS_10 signifies the role of signal transduction events and reactive oxygen species during early heat stress. The transcriptional profile shows that the response of rice to heat stress involves signal perception and transduction, activation/synthesis of heat shock factors, genomic and proteomic alterations, ROS metabolism and a host of other proteins associated with biochemical, cellular and physiological processes and ‘unknown’ functions. The synthesis of heat shock proteins (Hsps) appears an important response to heat stress. Genome-wide gene families of rice Hsp20, Hsp40, Hsp70 and Hsp100 as well as heat shock factors (Hsfs) were analysed. Rice ClpB/Hsp100 family proteins are constituted of three isoforms localized in three different cell compartments namely cytoplasm (ClpB-c), chloroplast (ClpB-p) and mitochondria (ClpB-m). Our present focus is to analyse the role of ClpB/ Hsp100 proteins in governing heat stress response in Arabidopsis and rice plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 25 Day 1: December 11, 2015
Concurrent Session A-II
Development of physiological and biochemical markers for monitoring forest health in a changing climate
Rakesh Minocha1*, Stephanie Long1, Swathi Turlapati2,1 and Subhash C. Minocha2 Rakesh Minocha 1USDA Forest Service, NRS, Durham, NH 03824, USA 2Department of Biological Sciences, University of New Hampshire, Durham, NH 03824, USA *Correspondence: [email protected]
Continuous input of pollutants from atmospheric deposition can move ecosystems towards soil Al mobilization/toxicity, nutrient depletion, heavy metal toxicity, and finally nitrogen saturation. A major goal of our research is to determine the usefulness of selected polyamines, amino acids, and phytochelatins as early metabolic markers for assessing forest productivity using asymptomatic forest stands. This type of early evaluation is beneficial in developing risk assessment and stress remediation strategies. Our objective is to determine the usefulness of polyamines, particularly putrescine, amino acids (e.g. arginine, Proline and GABA), phytochelatins (inducible by heavy metal toxicity) as foliar indicators of abiotic stress in visually asymptomatic conifer and hardwood trees. This presentation will be a review of several studies conducted over two decades in collaboration with scientists from various institutions using hardwood, conifer, and mixed wood study sites across Northern New England, USA over two decades. These data show a strong correlation between soil nutrient, these metabolites, and chlorophyll indicating that they can potentially be used as reliable and easily quantifiable markers of abiotic stress in visually healthy trees. Most of these select metabolites increased in response to exposure to increased levels of pollutants. In conjunction with these changes, changes in soil amino acids and microbial population diversity and functions were also observed. In some cases, such as Harvard Forest, MA, USA and Bear Brook Watershed, ME, USA, long-term ecosystem research (LTER) study sites these changes accompanied lower site productivity in red pine and red spruce stands, respectively.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 26 Day 1: December 11, 2015
Concurrent Session A-II
Tailoring rice plants for higher yield via genetic manipulation for ion partitioning, cellular detoxification and cytokinin metabolism
Khirod K. Sahoo1, Brijesh K. Gupta1, Rohit Joshi1, Amit K. Tripathi1, Ashwani Pareek2 and Sneh Lata Singla-Pareek1* Sneh Lata Singla-Pareek 1Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India 2Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Correspondence: [email protected]; [email protected]
To meet the rising food demand, there is an urgent need to boost crop production. Thus, we need to accelerate our understanding of reduction in crop yield occurring due to various climatic vagaries. Tremendous progress has been made over the past years in defining the molecular circuitry underlying cross talk between various plant hormones, defence related machineries, development associated proteins and environmental response networks. Here, we have attempted to bridge the yield gap in rice for sustainable food production by genetic manipulation of either stress related genes or genes participating in hormone homeostasis. Taking clues from the stress tolerance capacity of plants overexpressing either the glyoxalase (Gly) pathway for enhanced cellular detoxification or sodium proton antiporters (NHX as well as SOS1), we developed transgenic riceplants expressing two different stack combinations of stress associated genes viz. GlyI, GlyII along with NHX1 or with SOS1. Evaluation of these multigenic transgenic plants via assessment of various morphological, physiological and agronomical parameters indicated improved tolerance to abiotic stress and enhanced yield. Simultaneously, we also raised transgenic rice plants with RNAi based knockdown of one of the key genes of cytokinin metabolism i.e., Cytokinin oxidase-2. These transgenic plants showed better yield under both non-stress and stress conditions. Considering the complexity of stress response, our results suggest that a combinatorial approach targeting diverse pathways is better than the ‘single gene approach’ to tame the deleterious effects of environmental extremes of high salinity and water deficit under field conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 27 Day 1: December 11, 2015
Concurrent Session A-II
Transcriptome analysis under temperature stress in seabuckthorn, an important high altitude medicinal plant
Saurabh Chaudhary1, Rajesh Ghangal2, Ankit Jain3, Ashish Kumar1 and 1 Prakash C. Sharma * Prakash C. Sharma
1University School of Biotechnology, Guru Gobind Singh Indraprastha University, New Delhi, India 2Department of Biotechnology, Manav Rachna International University, Faridabad, India 3Centre of Excellence in Genomics, ICRISAT, Patancheru, Hyderabad, India *Correspondence: [email protected]
Seabuckthorn (Hippophae rhamnoides L.), a high altitude shrub, is known for its immense medicinal, nutritional and ecological value. The remarkable adaptation of seabuckthorn to harsh environments exhibited by tolerance to abiotic stresses specifically extreme temperatures has recently attracted attention of researchers as a non model important plant species. However, little is known about gene regulation and signalling pathways involved in extreme temperature (low and high) stress responses in this species. We analysed expressed sequence tags (ESTs) data followed by whole transcriptome sequencing from leaf and root tissues of sea buckthorn to explore the plant responses at molecular level under extreme temperatures. Moreover, to a gain deep insight into the molecular mechanism under cold and freeze stress we performed digital gene expression analysis using DeepSAGE analysis. Initially, 3412 ESTs were generated and assembled into 1665 unigenes, followed by assembly of 86,253,874 high quality NGS short reads into 88,297 putative unigenes. Next, from DeepSAGE data, 11,922 differentially expressed genes (DEGs) including 6539 up regulated and 5383 down regulated genes were identified under cold and freeze stress. Among various DEGs, 38 genes were validated in response to cold and freeze stress, using qRT-PCR. Recently, we have identified and validated 11 more genes, found to be differentially expressed under heat, cold and freeze stress simultaneously, suggesting a crosstalk between different stress pathways. The seabuckthorn transcriptome data have also been used to mine microsatellite markers for diversity analysis and transcription factors. The ample amount of data generated is under further analysis for gene discovery, development of functional molecular markers and other by products.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 28 Day 1: December 11, 2015
Concurrent Session A-II
Development of salt tolerant varieties following conventional, physi- ological and molecular approaches
Parbodh C. Sharma1*, S.L. Krishnamurthy1, Neeraj Kulshreshtha2, Jogendra Singh1, 1 1 Preeti Pundir and Dinesh Kumar Sharma Prabodh C. Sharma
1Central Soil Salinity Research Institute, Karnal-132001, Haryana India 2Sugarcane Breeding Institute RRS, Karnal-132001, Haryana, India *Correspondence: [email protected]
Among abiotic stresses, salt stress is the most complicated problem posing a major challenge. Around 1000 m ha globally and 6.73 m ha in India are affected by salinity. Central Soil Salinity Research Institute (CSSRI) has developed 15 salt tolerant varieties of different crops; 7 of rice (CSR10, CSR13, CSR23, CSR27, CSR Basmati 30, CSR36 and CSR43), 4 of wheat (KRL1-4, KRL 19, KRL210 and KRL213), 3 of Indian mustard (CS52, CS54 and CS56) and one in chickpea (Karnal Chana 1) through conventional breeding approaches to increase the productivity of salt affected soils for resource poor farmers. Physiological and biochemical approaches have been used to identify different mechanisms of salt tolerance and used for identification of salt tolerant and sensitive lines. The molecular approaches are also being used to develop salt tolerant rice varieties. Introgression of Saltol QTL to two mega high yielding rice varieties namely, Pusa 44 and Sarjoo 52 carried out through Marker Assisted Selection Back Cross (MABC) breeding. The varieties namely, Pusa 44 and Sarjoo 52 were selected as recurrent parents while FL478 was used as a donor parent and produce F1 seeds. True F1s were selected by the Saltol marker RM8094. Further backcrossing with respective recurrent parent and selfing was performed to produce seeds up to BC3F3 generation. In every back cross generation, recombinant and foreground selection was carried out. Desirable plants were selected based upon the (genotyping) marker data and through stringent phenotyping as similar to recurrent parent. Background selection and DUS characterization of NILs is in progress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 29 Day 1: December 11, 2015
Concurrent Session A-II
Introgression of physiological traits to save water while improving the productivity in rice
M.S. Sheshshayee1*, M.D. Prathibha1, M.V. Mohan Kumar1, B.R. Raju1, H.G. Jalendra 1 2 1 Kumar , M.P. Rajanna and M. Udayakumar M.S. Sheshshayee
1Department of Crop Physiology, UAS, GKVK, Bangalore-560065, India 2ZARS, VC Farm, Mandya-571401, Karnataka, India *Correspondence: [email protected]
Saving water and sustaining productivity under water limited conditions are the most important challenges. Global research strongly suggests an analytical breeding approach in lieu of the empirical breeding adopted presently. Thus, improving plant traits associated with maintenance of tissue water relations and metabolism have paramount significance. While scientists equivocally agree on the role of root traits in maintenance of water relations, there are debates on the relevance of WUE as a drought adaptive trait. We provide strong experimental evidences that emphatically prove the relevance of WUE as a potential trait for crop improvement under water limited environments. Further, introgression of several of such drought adaptive traits seems to be essential for a comprehensive improvement in productivity under water-limited conditions. We adopted conventional as well as an elaborate marker assisted backcross breeding (MABC) approaches to introgress WUE and water relation traits into a single elite genetic background. The progeny developed by these approaches led to an increase of yield by more than 15% combined with an ability to save at least 60% of irrigation water. The results of these investigations emphatically demonstrated the relevance of improving physiological traits by breeding.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 30 Day 1: December 11, 2015
Concurrent Session B-II
Signaling in plant immunity
Dierk Scheel*
Leibniz Institute of Plant Biochemistry, Department of Stress and Developmental Biology, Weinberg 3, D-06108 Halle (Saale), Germany Dierk Scheel *Correspondence: [email protected]
Plants detect potential pathogens in their environment via pathogen-associated molecular patterns (PAMPs) that are recognized by plant plasma membrane receptors. Typical PAMPs include the bacterial flagellin-derived flg22 peptide, the elf18 peptide of the bacterial elongation factor EF-Tu, bacterial peptidoglycans and lipopolysaccharides, as well as fungal chitin oligomers and glucan fragments from oomycetes. PAMP-binding to their receptors initiates complex signaling networks that activate a multi-component defense response and thereby establish PAMP-triggered immunity. One of the earliest detectable responses after PAMP perception is the activation of ion channels at the plasma membrane. Using a transgenic Arabidopsis line with the calcium reporter, aequorin, increases in cytosolic calcium levels are detected after PAMP application. To identify signaling network components, seeds of aequorin-expressing lines were mutagenized and the population screened for mutants with changed calcium elevation (CCE) in response to different PAMPs. Among several receptor complex components, a lectin S-domain receptor kinase was identified, which mediates lipopolysaccharide sensing in Arabidopsis thaliana. MAPK cascades are essential for controlling defense responses. The elements that prevent erroneous signaling crosstalk may include expression patterns of the MAPK cascade components, the presence of pathway-specific protein complexes or the MAPK substrate diversity. Different strategies have been employed to isolate MAPK interacting proteins. Several VQ-motif containing proteins are MAPK substrates and regulate immune responses.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 31 Day 1: December 11, 2015
Concurrent Session B-II
Natural variation and network modelling approaches to identify novel regulatory genes affecting plant development and water use
Ulrike Bechtold*, John Ferguson and Phil Mullineaux Ulrike Bechtold Plant Molecular Physiology Lab, University of Essex, School of Biological Sciences, Colchester, Essex, United Kingdom *Correspondence: [email protected]
The improvement of plant productivity while reducing water use is an urgent objective to sustain or improve plant production while conserving natural resources. To identify and exploit genes that promote yield in water limiting environments is an important challenge in plant biology and requires genes that affect lifetime traits such as water use. In their natural environment, plants may experience continuous or episodic drought stress, and we aim to understand how different types of drought stress impact on plant growth. In order to do this we obtained a quantitative measure of the natural genetic variation of yield and water use in 46 Arabidopsis thaliana accessions. Water use, developmental and physiological traits were assessed during episodic- and continuous drought experiments to evaluate if proxy traits for productivity and water use can be identified. Simultaneously we are using high-resolution time series transcriptome profiling coupled with Bayesian modeling to infer regulatory networks linked to early events of an episodic drought stress in order to identify key regulatory genes. Using these approaches we identified AGAMOUS- LIKE 22 (AGL22) as key gene in a regulatory transcription factor network. AGL22 is involved in the transition from vegetative state to flowering, and loss of AGL22 expression affected photosynthesis and water use providing a link between early changes in primary metabolism and the subsequent initiation of responses to stress that govern plant productivity. The combination of genomic, transcriptomic and systems biology approaches holds great promise to increase the efficiency of identifying novel candidate genes under different drought stress conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 32 Day 1: December 11, 2015
Concurrent Session B-II
Deregulated symbiosis receptor kinase hyperactivates spontaneous nodulation
Maitrayee Dasgupta*, Sudip Saha, Ayan Dutta and Abhishek Bhattacharrya Maitrayee Dasgupta Department of Biochemistry, Calcutta University, Kolkata, West Bengal, India *Correspondence: [email protected]
Plants restricted to a monophyletic clade of four angiosperm orders is capable of undertaking intracellular root nodule symbioses with nitrogen-fixing bacteria. A compatible interaction triggers the ‘Symbiosis response pathway’ (Sym pathway) to initiate the genetic reprogramming required for development of nodule primordia and intracellular colonization of symbionts in the newly divided cells. Gain of function of ‘Sym pathway’ members like CCaMK, CYCLOPS or its downstream effectors like HK1 can uncouple nodule organogenesis from bacterial infection and cause development of empty spontaneous nodules in the absence of the bacterial symbiont. The efficiency of spontaneous nodulation under constitutive activity of these members was either same or lower than the rhizobia induced nodulation, which meant that pathways restricting nodule numbers were functional under these conditions. Symbiosis Receptor Kinase (SYMRK), is an upstream receptor kinase of the ‘Sym pathway’. Our efforts have shown that ligand independent deregulated activation of the intracellular kinase domain of SYMRK triggered hyperactivation of nodule organogenesis. The merged clusters of spontaneous nodules indicated that constitutive SYMRK had overcome the control over nodule numbers or their spatial position and suggested the importance of SYMRK ectodomain in restricting nodule number. In presence of the symbiont the unrestrained kinase activity of SYMRK supported the root hair curls and epidermal infection threads but rhizobial colonization in the nodule interior was highly compromised in the absence of ectodomain of SYMRK. Profuse spontaneous nodulation triggered by constitutive SYMRK would be discussed in the backdrop of prior arts.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 33 Day 1: December 11, 2015
Concurrent Session B-II
Regulation of plant stress responses and development by the ROS- Ca2+ signaling network
Kazuyuki Kuchitsu1,2*, Kenji Hashimoto1, Hidetaka Kaya1 and Nobutaka Kitahata1 Kazuyuki Kuchitsu 1Department of Applied Biological Science; 2Imaging Frontier Center, Tokyo University of Science, Noda, Chiba 278-8510, Japan *Correspondence: [email protected]
Reactive oxygen species (ROS) are highly toxic molecules produced during photosynthesis and aerobic respiration. However, ROS are enzymatically produced in a highly regulated manner spatially and temporally. ROS play key roles in regulating a broad range of physiological processes, such as growth and development including tip growth of root hairs and pollen tubes, defense responses against biotic and abiotic stresses as well as cell wall metabolism. Respiratory burst oxidase homologues (Rboh) have been identified as ROS-producing NADPH oxidases, which act as key signalling nodes integrating multiple signal transduction pathways in plants. Rbohs are synergistically activated by Ca2+ binding to the N-terminal EF-hand motifs and phosphorylation by several families of protein kinases. We have identified several proteins that interact with the N-terminal cytosolic region of Rbohs in vivo and regulate their ROS-producing activity. Tip growth and ROS accumulation in pollen tubes were severely impaired in the rbohH rbohJ double mutant. Point mutations in the EF-hand motifs impaired Ca2+-induced ROS production and complementation of the double mutant phenotype, indicating that Ca2+-activated ROS production is essential for the proper tip growth. The ROS-Ca2+ positive feedback regulation seems to be conserved not only in polarized cell growth but also various signaling processes in plants. We recently identified and characterized two Rbohs from an emerging model liverwort Marchantia polymorpha. Evolutionary aspects and molecular regulatory mechanisms of Rboh/ROS-mediated signaling will be discussed. Furthermore, novel chemical screening led us to identify potential compounds that affect Rboh-mediated signaling events including plant defense responses positively and negatively.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 34 Day 1: December 11, 2015
Concurrent Session B-II
Cytosine DNA methyltransferases and components of PRC1 complex regulate growth, development and abiotic stress tolerance in the moss, physcomitrella patens
Meenu Kapoor*, Vimala Parihar and Deepshikha Arya Meenu Kapoor
Guru Gobind Singh Indraprastha University, New Delhi, India *Correspondence: [email protected]
Liverworts, hornworts and mosses form a paraphyletic group among bryophytes that diverged early from the lineage that gave rise to flowering plants. Among these, mosses have had a tremendous impact on different ecosystems and climatic conditions during the pre-cambrian era that paved way for colonization of land by animals. Physcomitrella patens occupies a key evolutionary position among bryophytes linking the green algae and the vascular plants. Its genomic study provides insight into the adaptations to extreme conditions that eventually lead to colonization of land by plants. The green yeast™, as it is sometimes called, exhibits a high rate of homologous recombination. This, together with the ease of transformation and high regeneration capacity has resulted in emergence of P. patens as a model plant for reverse genetics approaches for gene functions analysis. Exploiting this potential, we have generated stable gene knockout mutants for cytosine DNA methyltransferases, PpCMT and PpDNMT2 and the PRC1 component, PpLHP1. Whole genome transcriptome analysis has revealed the role of epigenetic modifications, such as DNA methylation, in regulating genes/biological pathways affecting resilience and abiotic stress tolerance. We provide genetic evidence for regulation of cell growth, cell wall loosening and gravitropic response by the PpCMT/PpLHP1 complex and for PpLHP1 affecting transition from juvenile to adult leafy gametophore stage. Protein-protein interaction studies reveal other molecular components involved in these pathways. We also show that the DNA/RNA methyltransferase, PpDNMT2, regulates salt and osmotic stress tolerance by affecting biogenesis/stability of specific transfer RNA molecules.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 35 Day 1: December 11, 2015
Concurrent Session B-II
Ethylene receptors and molecular mechanism of ethylene sensitivity in tomato
Ajay Arora* Ajay Arora Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Correspondance: [email protected]; [email protected]
The past two decades have been rewarding in terms of deciphering the ethylene signal transduction and functional validation of the ethylene receptor and downstream genes involved in the cascade. Our knowledge of ethylene receptors and its signal transduction pathway provides us a robust platform where we can think of manipulating and regulating ethylene sensitivity by the use of genetic engineering and validating through transgenics. Recently, major progresses have been made in understanding of the molecular mechanism regulating ethylene biosynthesis, perception and signal transduction pathways. In important ornamental plant species, we aim to identify homologues to genes involved in ethylene signaling, which previously have been isolated in model plants. Key genes are identified and their regulation is studied in the postharvest context. An alteration of ethylene action is a valuable target for the genetic engineering of crops. Manipulation of ethylene biosynthesis or perception allows us to modulate these processes and thereby create plants with more robust and desirable traits, giving us a glimpse into the role of ethylene in the plant. We isolated, characterized and transformed the gladiolus ethylene receptor and developed the tomato transgenics that confer reduced sensitivity to ethylene. The possible strategies for altering the ethylene sensitivity of tomato plants using ethylene receptor genes will be discussed.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 36 Day 1: December 11, 2015
Concurrent Session D
Metabolic engineering of pathways for enhancing oil and seed yield in Camelina sativa
Om Parkash Dhankher1*, Sudesh Chhikara1, Hesham Abdullah1, Kenny Ablordeppey1, 1 2 Parisa Akbari and Danny Schnell O.P Dhankher
1Stockbridge School of Agriculture, University of Massachusetts, Amherst, MA 01003, USA 2Department of Biochemistry and Molecular Biology, Univ. of Massachusetts, Amherst, MA 01003, USA *Correspondence: [email protected]
Liquid transportation fuels based on plant seed oils have tremendous potential as an alternative to petroleum-derived fuels. Camelina sativa, a member of brassicaseae, has been proposed as an ideal non-food crop for biofuel production. Increasing seed oil yield by manipulation of the triacylglycerol (TAG) biosynthesis pathway and by increasing the flux of carbon from increased photosynthesis to TAG synthesis will make this crop highly profitable. In seeds, TAG is synthesized from glycerol-3-P and fatty acids through the sequential activities of the glycerol-3-P acyltransferase, lysophosphatidic acid acyl transferase and diacylglycerol acyltransferase. Overexpression of enzymes that catalyze the synthesis of glycerol backbone and conjugation of fatty acids to glycerol backbone appear to be a promising target for increasing TAG accumulation. Therefore, to increase the oil contents and seed yield in C. sativa, we targeted the over expression of several genes involved in fatty acids synthesis and acylation of fatty acids in the downstream process for TAG synthesis. Over expression of genes involved in TAG synthesis resulted in a significant increase in seed and oil yield. Since C. sativa is fairly a new oil seed crop and information regarding genes that regulate the fatty acids and TAG biosynthesis remain poorly understood. It is not known how these genes contribute to the overall accumulation of TAG in the seed oil and what are the rate limiting steps in the biosynthesis of these molecules during seed development. Therefore, to identify the bottlenecks for further improving the seed and oil yield in C. sativa seeds, a comprehensive transcriptome and metabolome analysis was performed in wild type and transgenic C. sativa over expressing TAG related genes. Both transcriptome and metabolome profiling revealed several genes/ gene networks controlling the synthesis of key metabolite for TAG biosynthesis and storage during seed development. Efforts are underway to combine the expression of these key genes using gene-stacking approaches for further enhancing the oil and seed yield.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 37 Day 1: December 11, 2015
Concurrent Session D
Terpenoid sub-class based discrimination between MVA and DOXP pathways of isoprenogenesis in plants: Exceptional results to conceptual revision
Rajender Singh Sangwan* R.S. Sangwan
1Center of Innovative and Applied Bioprocessing (A National Institute under DBT, Govt. of India), C-127, Phase-8, Industrial Area, S.A.S. Nagar, Mohali-160071, Punjab, India *Correspondence: [email protected]; [email protected]
Biosynthesis of terpenoids in plants differs from that in animals by its recruitment of two independent routes of isoprenogenesis: the classical cytosolic mevalonate (MV) pathway and the recently discovered plastid localized deoxy- xylulose phosphate (DOXP) pathway. Redundancy of routes to generate isopentenyl pyrophosphate (IPP) and its isomer (dimethyl allyl pyrophosphate, DMAPP) invokes the issue of their relative significance. Early studies have indicated that sesqui-, tri- and polyterpenes are synthesized through MVA pathway while biosynthetic pathways for of mono-, di- and tetraterpenes recruit DOXP pathway generated IPP and DMAPP. Very recently, exceptions to such strict discrimination, based on sub-class of terpenoids, with respect to recruit of isoprenogenic route have appeared including observations from our on studies on triterpene ancestry specialized metabolites (withanolides) of Ashwagandha (Withania somnifera). Our studies have revealed that withanolides, despite being of triterpene origin, are synthesized with the involvement of substantial participation of DOXP pathway. Similarly, even differentially role of MVA and DOXP pathways has been encountered by some researchers for same the sub-class of terpenoids produced in different parts of the plant. While an article cites this phenomenon as “exception is the rule”, we have hypothesized a new conceptual line of biosynthetic load-driven model of relative recruitment of DOXP and MVA pathways for biosynthesis of isoprenoids (Plant Cell Reports 31: 1889-1897, 2012). An updated synthesis of this subject would be presented at the conference along with its implications in inter-organelle coordination of metabolic events and its applications in metabolic engineering for enhanced production of terpenoidal secondary metabolites.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 38 Day 1: December 11, 2015
Concurrent Session D
Enhancement of therapeutic molecules in medicinal plants using metabolic engineering approach
M.Z. Abdin* M.Z. Abdin Centre for Transgenic Plant Development, Department of Biotechnology, Faculty of Science, Jamia Hamdard, Hamdard Nagar, New Delhi-110062, India *Correspondence: [email protected]
Plants synthesize a vast array of secondary plant metabolites through a network of complex metabolic pathways that are regulated by endogenous factors and environmental conditions. In 1988 the NAPRALERT data base contains more than 88000 natural compounds, most of which were derived from plants. Though these compounds are believed to be non essential for the plant to live, they play an important role in the interaction of the plant with its environment ensuring the survival in its ecosystem. The plant secondary metabolites however, are of immense economical importance due to their use as flavouring agents, fragrances, insecticides, pesticides, dyes and drugs. Of all the drugs used in western medicine, about 25% are derived from medicinal plants, either as pure compounds or as natural synthons. The drugs derived from the medicinal plants include morphine, codeine, paclitaxel, vinblastin, vincristine, scopolamine, atropine, pilocarpine, physostigmine, digoxin, artemisinin etc. Best example of natural synthon is the steroid skelton, which is the basis of large series of drugs, such as contraconceptives and corticosteroids. The concentration of these compounds in medicinal plants is however, very low, for example the concentration of artemisinin in Artemisia annua L. is @1.0 %, while the concentration of curcumin in Curcuma longa is @6.0%, limiting their commercial exploitation. The metabolic engineering approach of secondary metabolic pathways therefore, has been used by us and other researchers to enhance the concentration these therapeutic molecules in medicinal plants so as to improve their commercial utilization.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 39 Day 1: December 11, 2015
Concurrent Session D
Secondary plant product pathway engineering for improved crop productivity and human health
Prabodh Kumar Trivedi* P.K. Trivedi CSIR-National Botanical Research Institute, Rana Prata Marg, Lucknow-226001, India *Correspondence: [email protected]; [email protected]
Plants have developed various approaches to magnetize pollinators, shielding themselves against UV-light and for their defence. One of the strategies adopted by plants for these purposes is based on synthesis of secondary metabolites, including flavonoids. Flavonoids participate in number of processes in plants as well as are beneficial for human health. Biosynthesis and accumulation of these secondary plant products is species-specific and under tight spatial and temporal regulation of gene expression. Therefore, number of food products consumed by human is deficient in these phytoceuticals. Due to diversity of secondary transformations and complexity of regulation of this pathway, single gene strategies have not been much successful in enhancing the flavonoid biosynthesis in target plants. We used transcription factors, carried out gene pyramiding as well as used artificial miRNAs to engineer flavonoid biosynthesis in plants. Analysis of tobacco and tomato transgenic lines revealed that use of specific transcription factors can lead to flux availability for phenylpropanoid pathway in general and flavonol biosynthesis in particular. The transgenic lines with enhanced flavonols developed resistance against the insect pests. Transgenic tobacco lines co-expressing MYB transcription factor and isoflavone synthase gene accumulated substantial amount of isoflavonoid being at the highest levels that could be engineered in non-leguminous plants. In addition, flavonoid rich tobacco and tomato showed value addition in relation to improvement in bone health in animals. Our results establish an efficient strategy for successful pathway engineering of flavonoids which can be used for improving medicinal value as well as insect resistance in plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 40 Day 1: December 11, 2015
Concurrent Session D
Enhancing nutritional value of oilseed Brassica juncea through genetic manipulation of glucosinolate biosynthesis and transport
Naveen K. Bisht* and Rehna Augustine Naveen K. Bisht National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Correspondence: [email protected]
Glucosinolates are Capparales-specific secondary metabolites that have immense potential in human health and agriculture. Brassica juncea, a globally important oilseed crop, contains relatively high amount of seed glucosinolates ranging from 80-120 μmol/g dry weight (DW). One of the major breeding objectives in oilseed Brassicas is to improve the seed-meal quality through the development of low seed glucosinolate lines, as high amounts of certain seed glucosinolates are known to be anti-nutritional and reduce the meal-palatability. Unlike for Arabidopsis thaliana, our knowledge about the complex genetical and biochemical regulation of glucosinolate biosynthesis in Brassica crops is sparse. Our laboratory have identified multiple homologs of key glucosinolate pathway genes (MAM1, CYP83A1, AOP2), and its transcriptional regulators (MYB28) from B. juncea and assessed the effects of allopolyploidization on divergence of gene sequence, structure, expression and function. Subsequently we found that, targeted silencing of BjuMYB28, a key transcriptional regulator, provided significant reduction of the anti-nutritional aliphatic glucosinolates fractions (ca. 11.26 μmol/g DW of seed), without altering the desirable non-aliphatic glucosinolate pool. The low glucosinolate trait was stable in subsequent generations of the transgenic lines with no visible off-target effects on plant growth and development, when tested under containment field conditions. Enrichment of glucoraphanin, a glucosinolate known for its potent anti-carcinogenic properties, is another area of ongoing research in our laboratory. Besides, a part of my presentation will discuss selective manipulation of glucosinolate transport processes, which will be a favourable strategy to engineer the low glucosinolate trait in oilseeds without compromising the fitness of plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 41 Day 1: December 11, 2015
Concurrent Session D
Changes in metabolites and defense related enzymes in groundnut during leaf spot diseases
Mahesh Kumar Mahatma1*, Sujit Kumar Bish1, Lokesh Kumar Thawait1, Kuldeep Singh 1 1 2 2 Jadon , Nilesh Khatediya , Khyati J. Rathod and B.A. Golakiya M. K. Mahatma
1ICAR-Directorate of Groundnut Research, Post Box No. 5, Junagadh-362001, Gujarat, India 2Department of Biotechnology, Junagadh Agricultural University, Junagadh-362001, Gujarat, India *Correspondence: [email protected]
Leaf spots are major diseases affecting groundnut productivity worldwide. Early and late leaf spot diseases are caused by two distinct, but closely related fungi. Late leaf spot (LLS) is generally more aggressive and difficult to control than early leaf spot. In the present investigation LLS disease resistant (AK-265 , ICGV-86590, FDRS-10, GPBD-4 and CS-186) and susceptible genotypes (JL-24, JAL-42, GG 20, GG-7 and TG37A) were sown in the pots during Kharif 2014 and leaf samples were taken from control and infected plants at 45 and 90 days after sowing. Sugars and phenol were separated and analyse by Ion chromatography and LC-MS/MS, respectively. Glucose and sucrose content increased in resistant genotypes whereas decreased in susceptible genotypes after infection. These results suggested that the sugars might be utilized by the pathogen for growth and development. Inositol content was increased in all genotypes after infection. Higher constitutive and induced level of salicylic acid was observed in resistant genotypes. Resistant genotypes had about 2-3 times higher constitutive level of salicylic acid. Moreover, resistant genotypes also had appreciably higher content of p-coumaric acid than that of susceptible genotypes at infected and uninfected stages. Peroxidase and polyphenols oxidase activities were also higher in resistant genotypes. Thus, results revealed that higher sugars in resistant genotypes may provide higher amount of substrate for phenylpropanoid pathway which leads to synthesis of higher content of salicylic acid and coumaric acid and thus confers disease resistance in groundnut
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 42 Day 2: December 12, 2015
Plenary Session-II
Rice genome for forward and reverse genetics
Akhilesh K. Tyagi1*, S.K. Parida1, S. Das1, N. Malick1, S. Kapoor2, A.K. Singh3 and P. Agarwal1
1National Institute of Plant Genome Research, New Delhi-110067, India 2Department of Plant Molecular Biology, Univ. of Delhi, South Campus, New Delhi-110021, India Akhilesh K. Tyagi 3Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India *Correspondence: akhilesh @genomeindia.org
We have sequenced genome of rice as part of IRGSP ten years back. In the post-genomic era, significant progress has been made in transcriptomics and gene discovery by way of forward and reverse genetic approaches. Efforts are on to fill the gap between the genome and the phenotype. This may lead to regular practice of genomics-assisted molecular breeding. One of the primary aims of ongoing investigations in our group is to understand gene function, phylogeny and regulatory networks. Several general conclusions of great relevance emerging from these studies include: i. new genes and taxon-specific clades can be identified by using molecular model based comparative genomics, ii. signatures of co-evolution in interacting proteins are discernible, iii. duplicated genes have undergone neo-/sub-/pseudo-functionalization, iv. highly significant number of genes is differentially expressed in spatial/temporal manner and in response to stress, and v. such analysis provides useful input for selecting target genes/promoters for functional genomics. At the same time investigations on defining the role of seed-specific genes by reverse genetics and identification of QTLs/genes by genetic and association mapping as well as by haplotyping have been undertaken. As we integrate molecular biology to genetic enhancement activity and overcome practical impediments in taking the knowledge from lab to land, we hope to reap the benefit of genomics research for plant improvement.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 43 Day 2: December 12, 2015
Plenary Session-II
Natural variation in plant circadian clocks
C. Robertson McClung1*, Richard M. Amasino2, Brent E. Ewers3, Kathleen Greenham1, Victor Hermand1, Ping Lou1, Todd C. Mockler4, Cynthia Weinig3,5, Scott T. Woody2 and Qiguang Xie1,6 C. Robertson McClung 1Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire, USA 2Department of Biochemistry, University of Wisconsin, Madison, Wisconsin, USA 3Department of Botany, University of Wyoming, Laramie, Wyoming, USA 4Donald Danforth Plant Science Center, Saint Louis, Missouri, USA 5Department of Molecular Biology, University of Wyoming, Laramie, Wyoming, USA 6Hebei Key Laboratory of Molecular and Cellular Biology, College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei, China *Correspondence: [email protected]
We have developed a high-throughput motion estimation system to measure circadian parameters by leaf movement. Analysis of a nested association mapping (NAM) panel from seven Recombinant Inbred Line (RIL) populations of Arabidopsis thaliana, supplemented with accessions from the 1001 Genomes Project, has identified multiple Quantitative Trait Loci (QTL) linked to known clock genes, as well as at least one novel QTL. Among natural populations of Mimulus guttatus and elite cultivars of soybean (Glycine max), we observe a strong latitudinal cline in circadian period in both species. This suggests that the clock contributes to performance in both natural and cultivated conditions. This is consistent with our growing understanding of the roles of the clock in the regulation of flowering and in the temporal coordination of growth and development, and in abiotic and biotic stress responses. In Brassica rapa, we have mapped QTL for circadian period, water use efficiency (WUE), shade avoidance, and other morphometric traits. There is striking colocalization of QTL for circadian period with QTL for shade avoidance and WUE. We have identified GIGANTEA (GI) as a major QTL responsible for natural variation in circadian period and have identified the causal nucleotide polymorphism responsible. This same nucleotide polymorphism underlies variation in hypocotyl elongation, cold and salt tolerance. Thus, natural variants of circadian clock genes such as GI offer attractive targets for the development of crops with enhanced stress tolerance and improved yield.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 44 Day 2: December 12, 2015
Plenary Session-II
Functional relevance of stress specific transcription factors bZIP60, MYC57 and NAC1 from Eleucine coracona in improving stress tolerance in rice
K.C. Babitha, S.V. Ramu, H.V. Ramegouda, Nataraja N. Karaba and M. Udayakumar* M. Udayakumar
Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus, Bangalore-560065, Karnataka, India *Correspondence: [email protected]
Moisture stress induced decrease in cell turgor affects several metabolic processes and amongst these, protein synthesis and protein turnover are the most challenged. Major stress induced constraints are translation processes and protein folding and stability. Stress specific transcription factor (TFs) might play a pivotal role in regulating the functional genes to sustain these processes. With this back ground, from the stress specific transcriptome of finger millet we cloned and characterized three stress specific TFs EcNAC1, EcMYC57, EcbZIP60 involved in regulating the expression of genes associated with protein folding and stability. These TFs are upregulated under diverse stress conditions. Overexpression of these TFs in tobacco improved tolerance to diverse abiotic stresses. The EcbZIP60 transformants showed tolerance to DTT and tunicamycin induced ER stress by regulating expression of relevant genes involved in protein folding. The EcMYC57 and EcNAC1 improved tolerance predominantly by upregulating the several dehydrins. Transgenic plants co-expressing of EcNAC1, EcMYC57, EcbZIP60 showed substantially improved tolerance to diverse abiotic stresses expressing ER stress responsive genes like chaperones (BiP, CRT, PDIL), dehydrins (LEA14, rd29A, rd29B), ABA signaling (PP2C). These results not only signify the relevance of genes associated with protein folding and stability in improving the stress adaptation but also provide evidences that the simultaneous expression of relevant genes has specific advantage over single gene transgenics. Based on this proof of concept the rice transgenic plants developed co-expressing EcNAC1, EcMYC57, EcbZIP60 showed improved tolerance to ER stress besides salinity and dehydration. Further these transgenics maintained higher spikelet fertility and increased productivity under semi irrigated aerobic conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 45 Day 2: December 12, 2015
Concurrent Session E
Marker assisted breeding for disease / pest resistance
Ramesh V. Sonti*
CSIR-Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad-500007, Telangana, India Ramesh V. Sonti *Correspondence: [email protected]
Marker assisted selection (MAS) is a very effective tool for crop improvement that involves use of variations in DNA sequences as flag posts to make selections of varieties that have new and advantageous combinations of genes. MAS can be applied for crop improvement when the trait of interest is present within the gene pool/extended gene pool of the crop of interest. I will present our collaborative work with the Indian Institute of Rice Research, Hyderabad, on development of a rice variety that is resistant to bacterial blight as a case study on how MAS can be gainfully used for crop improvement. This variety, called Improved Samba Mahsuri (ISM), is in commercial cultivation and is being appreciated by farmers in rice growing regions where fine grained varieties are cultivated and bacterial blight is a production constraint. I will present recent results which indicate that ISM exhibits tolerance to another bacterial disease called leaf streak; a disease which is gaining in importance and for which very few sources of resistance are available. Furthermore, we have developed an EMS mutagenized population of the elite rice variety Samba Mahsuri that we have used to isolate new mutants that are resistant to bacterial blight and other mutants that exhibit enhanced tolerance to yellow stem borer, a serious insect pest of rice. I will discuss our plans for further characterization of these mutants and how this work can be used to extend the scope of MAS.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 46 Day 2: December 12, 2015
Concurrent Session E
Maize defense signaling in response to aphid infestation
Joe Louis1,2*, Saumik Basu1 and Suresh Varsani1
1Department of Entomology; 2Department of Biochemistry, University of Nebraska-Lincoln, Lincoln, NE 68583, USA Joe Louis *Correspondence: [email protected]
Maize, one of the world’s significant cereal crops, suffers severe yield losses due to insect infestation. Corn leaf aphid [CLA; Rhopalosiphum maidis (Fitch)] is an economically important pest of several monocot crops, including maize. In addition to extensive crop damage, CLA also acts as a vector for viruses that cause devastating diseases in maize. The maize genotype, Mp708, developed by classical plant breeding confers enhanced resistance to CLA compared to B73 and Tx601 maize inbred lines. Feeding by CLA triggers the rapid accumulation of the Maize insect resistance1 (Mir1) transcripts, which encodes a cysteine protease. Since Maize insect resistance1-Cysteine Protease (Mir1-CP) is expressed in the vascular tissues, we examined the effect of vascular sap exudates by including it in the aphid diet. The number of aphids recovered from diet containing vascular sap collected from uninfested plants was comparable between Mp708 and Tx601 genotypes. However, if plants were infested with CLA prior to sap collection, Mp708 vascular sap significantly reduced the number of aphids compared to Tx601, suggesting that CLA-feeding induced accumulation of toxic factors, presumably Mir1-dependent factors, contribute to defense against aphids. Furthermore, our results demonstrate that recombinant Mir1-CP provides direct toxicity to CLA. Finally, we recently identified that foliar feeding by CLA rapidly sends defensive signal(s) to the roots that trigger belowground accumulation of the Mir1, signifying a potential role of long-distance signaling in maize defense against the phloem-feeding insects.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 47 Day 2: December 12, 2015
Concurrent Session E
Targeting of chitinase and cathepsin L genes of Helicoverpa armigera by RNAi confers insect resistance in tomato
Mamta and Manchikatla Venkat Rajam* M.V. Rajam Polyamine, Transgenic and RNAi Laboratory, Department of Genetics, University of Delhi South Campus, Benito Juarez Road, New Delhi-110021, India *Correspondence: [email protected]
Helicoverpa armigera, a polyphagous and notorious insect pest causes huge yield losses in many crop plants, and various insect control strategies met with limited success. Hence, novel alternate strategies are required to combat insect pests. RNA interference (RNAi), a sequence-specific gene silencing mediated by dsRNA emerged as a novel tool for insect control, where host plants are engineered to produce dsRNAs against vital genes of insect pests. In the present study, we have identified chitinase (HaCHI) and cathepsin L (HaCL) genes as potential targets for control of H. armigera as they are involved in insect molting, metamorphosis and survival. The off-target free partial sequences were selected from CDS and 3’UTR regions of HaCHI and HaCL genes respectively, and they were isolated and used for preparation of hair-pin RNAi constructs. Several RNAi tomato lines were generated by Agrobacterium- mediated transformation with these constructs. Continuous feeding of insect larvae on leaves of RNAi tomato lines reduced the larval growth, development and caused mortality. Various developmental deformities occurred in pupa and adults developed from larvae fed on leaves of RNAi lines. Reduction in fertility/fecundity was also observed in adults as compared to the control adults. The q-PCR analysis showed reduction in target gene transcripts in larvae fed on RNAi lines, and this could be correlated with the observed RNAi effects on insects. These results demonstrate that the chitinase and cathepsin L are essential for insect growth and development, and RNAi holds a great potential for effective management of H. armigera.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 48 Day 2: December 12, 2015
Concurrent Session E
Rep mediated transcription of geminivirus genome : Bringing in a new perspective
Nirbhay Kushwaha, Mansi Bhardwaj and Supriya Chakraborty* Supriya Chakraborty School of life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Correspondence: [email protected]
Geminiviruses are a group of plant viruses with ssDNA genome, which replicate in the nucleus of the infected cell. Replication initiator protein (Rep) encoded by a geminivirus is an early protein that is indispensible for viral replication. Although, multitasking roles of Rep in replication has been elucidated, information about its role in efficient gene expression is lacking. Chilli leaf curl virus (ChiLCV) is a monopartite geminivirus that can replicate in permissive host Nicotiana benthamiana. For efficient and orderly transcription from viral genomic DNA, recruitment of machineries responsible for histone post-translational modifications (PTMs) on viral promoter is essential. The present study evidently proves that deposition of histone 2B monoubiquitination (ubH2B) promotes tri-methylation of histone 3 at lysine 4 (H3K4me3) on ChiLCV promoter. We isolated homologues of two major components of monoubiquitination machinery, UBIQUITIN CONJUGATING ENZYME UBC 2 (NbUBC2) and HISTONE MONOUBIQUITINATION1 (NbHUB1) from N. benthamiana. TRV based virus induced gene silencing of NbUBC2 and NbHUB1 has indicated their crucial roles in ChiLCV pathogenesis. Rep redirects NbUBC2 from the cytoplasm to the nucleusand interacts with NbHUB1 and NbUBC2 in the nucleus. Furthermore, NbUBC2 and NbHUB1 silencing reduced deposition of H3K4me3 and ubH2B on host chromatin and down-regulated expression of host genes. Together, the current study reveals that ChiLCV Rep protein binds to the viral genome and recruits two host proteins NbUBC2 and NbHUB1. These proteins further help in monoubiquitination of histones 2B, and accumulation of H3K4me3 on viral mini- chromosomes, and in turn enhance transcription of the viral genome.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 49 Day 2: December 12, 2015
Concurrent Session E
Secreted effectors of necrotrophic fungi: Quest for possible host targets
Praveen Verma* Praveen Verma Plant Immunity Laboratory, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Correspondence: [email protected]
Recent studies on genomics of both plant and phytopathogens are beginning to unravel how these organisms manipulate host immunity to cause diseases. Plants recognize pathogens through strategies involving both conserved and variable pathogen elicitor, and pathogen manipulate the defense response through secretion of virulence effector molecules. However, only a few studies have explored for necrotrophic fungal pathogen effectors. Chickpea-Ascochyta rabiei system provides an excellent model for studying necrotrophy. We have sequenced 34.6 megabase draft genome sequence of Ascochyta rabiei and predicted 10,596 protein-coding genes that encode large and diverse inventories of secretory proteins, carbohydrate-active enzymes, transporters and primary and secondary metabolism enzymes, reflecting its necrotrophic lifestyle. Comprehensive analysis predicted a set of 758 secretory proteins with different conserved motifs. Further analysis of the secretome revealed a high abundance of PEXEL-like effector candidates. We characterized a novel PEXEL-like effector, PEC25, and found that this effector is indispensable for A. rabiei virulence. PEC25 translocate into the host nucleus during infection, which is critical for its functionality. To identify the host target, we explored the candidate genes of major QTLs of chickpea against A. rabiei resistance for PEC-25 interaction. Interestingly, we showed that PEC-25 is interacting with transcription factor present in QTLAR1 in the nucleus of chickpea. Our research will give new insights to elucidate the necrotrophic mode of infection through effectors and their targets.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 50 Day 2: December 12, 2015
Concurrent Session E
Plant-pathogen interaction with respect to plant parasitic nematodes: A sophisticated dialogue
Amar Kumar1*, Neha Shukla1, Pritam Kaur1, Rachita Yadav2, Arun Jagannath1, Shailendra Goel1 and Manu Agarwal1 and Ramneek Gupta2 Amar Kumar 1Department of Botany, University of Delhi, Delhi-110007, India 2Center for Biological Sequence Analysis, Technical University of Denmark, Denmark *Correspondence: [email protected]
Plants are continuously exposed to a wide range of pathogens, including plant parasitic nematodes. Being sessile, plants have developed elaborate innate defense mechanisms that allow them to recognize potential invading pathogens and to initiate successful defenses. I will initially present our current state of understanding of plant pathogen interactions followed by our own research on plant-nematode interactions. Plant endoparasitic nematodes (cyst and root-knot nematodes) are major pests of crop plants. Potato cyst (Globodera species) and root-knot (Meloidogyne species) are obligate biotrophs and takes between 3-6 weeks to complete their life cycle in the host roots. Roots are invaded by infective juveniles (J2) and develop feeding sites with highly modified host cells. Formation of feeding cells is activated by nematode secretions, which cause dramatic morphological and physiological changes in root cells and converts them into nutrient sinks. Earlier, we have cloned and characterized a tomato gene (Hero) that confers resistance against Globodera species by disrupting initiation and development of feeding cells in roots. Recently, we have taken genome-wide approaches by transcriptome and miRNAome analyses to investigate genes and regulatory miRNAs from tomato that are involved in disease development during root-knot nematode infection. We have shown that a set of genes (e.g., WRKY transcription factor, Cyclin D3-1, Putative Hydrophobic protein OSR8, Auxin Efflux carrier) and miRNAs (e.g., mi156, miR164, miR319) from tomato are up- and down-regulated at various stages of disease development and these are likely to play a crucial role in determining the outcome of plant-nematode interaction.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 51 Day 2: December 12, 2015
Concurrent Session F
Molecular response of plants to phosphate deficiency
K.G. Raghothama1*, V.K. Nagarajan1 and A. Jain2
1Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN-47907, USA K.G. Raghothama 2National Research Centre on Plant Biotechnology, Lal Bahadur Shastri Building, Pusa Campus, New Delhi-110012, India *Correspondence: [email protected]
Phosphorus, a key nutrient regulating metabolism and energy balance impacts a multitude of plant processes. Many of them are regulated at transcriptional and post transcriptional levels to enable plants to acquire, mobilize and recycle phosphate. Systemic and local responses of root system allow the plant to acquire phosphate from the rhizosphere. Cross talk with sugar, phytohormones and other nutrient signaling pathways is a part of phosphate starvation response mechanism. Pi deficiency modulates expression of Pi starvation-responsive (PSR) genes that are coordinately regulated by several transcription factors (TFs). Our research is focused on dissecting transcriptional regulation of phosphate starvation responses in Arabidopsis. Studies show that low Pi stress triggers expression of several transcription factors including Myb62, WRKY75, Zat6, HHO2 and ERF, E3 ligase among others. Analysis of these transcription factors using gene knockout and overexpression plants reveal a diverse role for them in regulating root system architecture, interaction with sugar, phosphate acquisition and utilization. Further Myb62 and ERF are also involved in GA mediated root responses to phosphate starvation. Studies revealed that subset of Pi starvation- induced (PSI) responses mediated by HHO2 is under regulation of PHR1 which is considered to be a major regulator of PSI. Transgenic plants with altered expression of transcription factors exhibit interesting root and shoot phenotypes, sensitivity to sugar and GA highlighting the complexity of plant response to phosphate deficiency. In spite of recent advances questions related to sensing and signaling, post transcriptional regulation, hormone and sugar interactions still need to be answered.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 52 Day 2: December 12, 2015
Concurrent Session F
Genome-wide expression of transcriptomes under iron and zinc stresses revealed candidate genes for kernel iron and zinc accumulation in maize (Zea mays L.)
Mallana Gowdra Mallikarjuna1, Nepolean Thirunavukkarasu1, Rinku Sharma1, H. S. Gupta Kaliyugam Shiriga1, Shikha Mittal1, Firoz Hossain1 and Hari Shanker Gupta2*
1Maize Research Lab, Division of Genetics, Indian Agricultural Res. Inst., New Delhi-110012, India 2Borlaug Institute for South Asia, CG Block, NASC Complex, New Delhi-110012, India *Correspondence: [email protected]
Deficiency of iron (Fe) and zinc (Zn) causes micronutrient-malnutrition or hidden-hunger affecting millions of people globally including sub-Saharan Africa, South Asia and Latin America where, maize is one of the major sources of food and feed. Among several available approaches, genetic-biofortification is a sustainable and cost-effective approach in addressing hidden-hunger. Identification of key genes involved in Fe and Zn homeostasis is an important prerequisite to breed micronutrient-rich hybrids. A transcriptome assay was performed, with ~18,000 transcripts distributed across the maize genome, in a SKV-616 inbred after exposing the seedlings to three Fe and Zn stress treatments (+Fe -Zn, -Fe+Zn, -Fe -Zn) along with control (+Fe+Zn). A total of 306 and 715 genes expressed in root and shoot, respectively were found common across stress treatments, suggesting presence of common transporters controlling both minerals in root and shoot. Differential expression of genes under stress treatments revealed that Zn association in many pathways was more pronounced over Fe pathways and the physiological demand for these minerals in shoot was much higher than in root. Significant expression of genes involved in mugineic acid pathway, metal-transporters, and phyto-hormone metabolisms revealed the role of those pathways in understanding the physiological and molecular mechanisms of Fe and Zn homeostasis. Co-mapping of DEGs with known QTLs for kernel Fe and Zn concentration identified several candidate genes such as OPT, metal-binding-protein, nramp3 and NAS. The candidate genes identified in the present investigation could be used for breeding kernel Fe- and Zn-rich maize hybrids.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 53 Day 2: December 12, 2015
Concurrent Session F
Identification of genomic regions and genes for nitrogen use efficiency in rice (Oryza sativa L.)
C.N. Neeraja*, S.R. Voleti, D. Subrahmanyam, K. Surekha, I. Subhakara Rao and B. Srikanth C.N. Neeraja Indian Institute of Rice Research, Hyderabad, Telangana, India *Correspondence: [email protected]
Nitrogen use efficiency in rice crop is crucial in climate change for reduction of nitrous oxide emission resulting from excess nitrogen fertilizer application and also in reduction of cost of cultivation. To identify genomic regions and genes for NUE in rice, various strategies viz., QTL and association mapping, micro array, real time PCR, transcriptome and micro RNA analyses were applied to germplasm and biparental mapping populations. Around 700 genotypes were screened under low and recommended nitrogen levels in field and wide genetic variability was observed. A set of genotypes was subjected to association mapping with 50 SSR markers and ~100 traits derived from field and hydroponics screening and 26 genomic regions found to be associated with more than 10 traits. Differential expression of PAL, OsSPL 14, Ammonium transporter, Glutamine synthetase and Nitrate Reductase was observed through qRT-PCR. Using genotypes with differential response under low N, applying Next Generation Sequencing strategies, several up and down regulated genes were analyzed in root and shoot tissues of two genotypes using microarray analysis; panicles of two genotypes were identified on HiSeq 2000 using 2x100 bp pair-end sequencing and using whole genome mRNA sequencing. The identified differentially expressed genes are being validated in the mapping populations and being introgressed into popular varieties.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 54 Day 2: December 12, 2015
Concurrent Session F
Comparative functional genomics of GPCRs and G-proteins: A new match-making service?
Nandula Raghuram1*, Navjyoti Chakraborty1, Kostya Kanyuka2, Ravi Pathak1, Devapriya 3 4 Choudhur and Richard Hooley Nandula Raghuram
1Molecular Biology and Functional Genomics Laboratory, University School of Biotechnology, Guru Gobind Singh Indraprastha University, Sector 16C, Dwarka, New Delhi-110078, India 2Department of Plant Biology and Crop Science, Rothamsted Research, West Common, Harpenden, Hertfordshire, UK 3School of Biotechnology, Jawaharlal Nehru University, New Delhi-110067, India 4Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath BA2 7AY, UK *Correspondence: [email protected]
Plant G-proteins and canonical G-protein coupled receptor(s) are best represented by Arabidopsis GPA1 and GCR1. But their partnership in plant signalling became controversial due to the competing all-or-none hypotheses. We used comparative functional genomics approach to analyse the whole transcriptomes of GPA1/GCR1 single and double mutants of Arabidopsis (gpa1-5, gcr1-5 and gpa1-5gcr1-5). By comparing the transcriptomes of the single mutants, we found that 30% of all GCR1-responsive genes and 57% of all GCR1-regulated processes were similar to those of GPA1, though there were also 246 DEGs and 43 processes that did not overlap with those of GPA1. Similarly, pairwise comparison of the differentially expressed genes (DEGs) from the double mutant with each single mutant revealed commonality of 115 or 33% of the 350 GCR1-regulated genes and 214 or 54% of the 394 GPA1 regulated genes respectively. When the double mutant was included in the comparison, though the common genes constituted only 6.2% of the 1,031 DEGs found in all 3 mutants, they represented 34% of all the 160 processes, indicating their co-regulation. This is by far the most compelling evidence, not only in favour of the GCR1-GPA1 partnership, but also in favour of its possible coexistence with other alternative partnerships, contrary to all-or-none hypotheses. The alternative partnerships may involve other GCRs, RGS1 or hitherto unknown GPAs. Our identification of the genes/ processes that could be regulated by such combinatorial partnerships provides a functional basis for match-making between different components of G-protein signalling for the first time.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 55 Day 2: December 12, 2015
Concurrent Session F
Foxtail millet: A climate change resilient model C4 crop with rich genetic and genomic resources for genetic improvement of cereals and bioenergy grasses
Manoj Prasad* Manoj Prasad
National Institute of Plant Genome Research, New Delhi-110067, India *Correspondence: [email protected]
Foxtail millet (Setaria italica L.) is recently regarded as a tractable experimental model crop for studying functional genomics of millets and bioenergy grasses. Further its potential abiotic stress tolerance has invited much research attention and we attempted to generate genomic resources and to decipher the molecular mechanism of stress tolerance. Though our efforts has generated considerable knowledge on structural and functional genomics of foxtail millet, the release of genome sequence significantly accelerated the genomic research to a new dimension, as evidenced by large-scale development of genomic resources, development of web-based database, genome-wide association studies, genome-wide functional genomics and standardization of molecular protocols specific to foxtail millet. Our research contributions in both the functional and structural genomic aspects of foxtail millet are remarkable and our efforts have now resulted in the transformation of this orphan crop into a nutri-crop with rich genomic and genetic resources. These resources would promisingly assist in crop improvement programs involving millets, non- millets and bioenergy grass species, thus ultimately addressing global food security.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 56 Day 2: December 12, 2015
Concurrent Session F
Comparative transcriptome and co-expression network analysis of two Brassica juncea cultivars in response to nitrate treatments
Parul Goel1,2, Rohit Chauhan1, Mohit Kumar Swarnkar1, Shivalika Pathania1, Vandna 1 1 1 1,2 Chawla , Vishal Acharya , Ravi Shankar and Anil Kumar Singh * Anil Kumar Singh
1Division of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061 (HP), India 2Academy of Scientific and Innovative Research, New Delhi-110012, India *Correspondence: [email protected]; [email protected]
Brassica juncea is an economically important oilseed crop of India. The Nitrogen use efficiency (NUE) of this crop is lower than other crops. We employed RNA-seq approach to understand transcriptional response of two contrasting genotypes Pusa Bold (high NUE) and Pusa Jaikisan (low NUE) of B. juncea in terms of nitrogen use efficiency under various nitrate treatments. A total of 42 RNA-seq libraries were sequenced using Illumina GAIIx that generated 446,580,418 paired-end reads which were utilized for de novo assembly following quality filtering. A total of 46,556 transcripts were generated with N50 1,393bp. A total of 20,884, 11,327 and 12,575 transcripts were annotated under GO, EC and KEGG, respectively. Our study showed that nitrate as a signal regulates expression of several genes involved in processes like signaling, nitrogen metabolism, carbohydrate metabolism, photosynthesis, secondary metabolite production etc. Moreover, genes involved in nitrate transport and assimilation were found to be induced more rapidly in Pusa Bold as compared to Pusa Jaikisan after nitrate treatment. Weighted gene co-expression network analysis of RNA-seq data of both the varieties was performed to explore their differential behavior in response to nitrate. From our network analysis, conserved modules in both varieties that respond to nitrate treatment were identified. Based on the comparison of shared genes, a HUB transcription factor mTERF was found to be differentially connected in both datasets in response to nitrate treatment. Through this study, we made an attempt to understand the molecular regulation of nitrate response in Brassica juncea.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 57 Day 2: December 12, 2015
Concurrent Session G
Structural biology of pre- and post-receptor regulation of auxin hormone responses
Joseph Jez*, Lucia Strader, Corey Westall, Ashley Sherp and David Korasick Joseph Jez Department of Biology, Washington University, St. Louis, MO, USA *Correspondence: [email protected]
Plants respond to developmental cues and abiotic and biotic stresses by controlling both the level and activity of various hormones, including auxins, jasmonates and salicyclic acid. Recent structural biology studies provide new insights on control mechanism before and after binding of auxin and jasmonates to their respective receptors. Pre- receptor action by acyl acid amido synthetases of the GH3 protein family modulate hormone levels by amino acid conjugation. The first crystal structures of benzoate-specific Arabidopsis thaliana AtGH3.12/PBS3 and jasmonic acid (JA)-specific AtGH3.11/JAR1, combined with biochemical analysis, define features for the conjugation of amino acids to diverse acyl acid substrates and highlight the importance of conformational changes in the C-terminal domain for catalysis. These results demonstrate how a highly adaptable three-dimensional scaffold is used for the evolution of promiscuous activity across an enzyme family for modulation of plant signaling molecules. A second molecular control mechanism in auxin responses involves protein-protein interactions mediated by the Phox/Bem1p (PB1) domains of Auxin Response Factors (ARF) and Aux/IAA co-repressors to regulate transcriptional events modulated by the phytohormone auxin. These structures reveal a two-pronged interaction and evolutionarily conservation of this core recognition feature, along with co-varying interface sequences allows for versatile PB1-mediated interactions in auxin signaling. Combined structural, functional, and sequence analysis provides new molecular insights that inform future inquiry into plant auxin responses.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 58 Day 2: December 12, 2015
Concurrent Session G
NPR1-dependent and –independent salicylic acid signaling in Arabidopsis
Ashis K. Nandi* Ashis K. Nandi School of Life Sciences, Jawaharlal Nehru University, New Delhi-1110067, India
*Correspondence: [email protected]
Amongst the plant hormones, salicylic acid (SA) plays the most significant role in defending biotrophic and hemibiotrophic pathogen. Plants accumulate SA upon pathogen accumulation, which is required for activation of defense. SA activates a transactivator protein NPR1 (Non-expressor of PR1), the master regulator of SA signalling. Several suppressors of npr1 mutation suggest the presence of NPR1-independet pathways for defense signaling, which remain elusive. Through screening of EMS mutagenized Arabidopsis plants, we identified cdd1 (constitutive defence without growth defect 1) as a suppressor of npr1. In contrast to other npr1 suppressors, cdd1 is not compromised for growth and development. Further, through transcriptome analysis of cdd1, we identified and characterized a novel zinc-finger domain containing protein ZFD1 that positively regulates defense signalling. Pathogens or SA induce ZFD1 expression. Through screening of Arabidopsis cDNA library in a yeast-2-hybrid assay, we identified a large numbers of interactors that provide mechanistic support for ZFD1 function. The epistatic interaction studies with npr1 mutant suggest that ZFD1 influences both NPR1-dependent and –independent pathways of SA signaling.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 59 Day 2: December 12, 2015
Concurrent Session G
Biotechnological method of production of orchid tubers containing biologically active substances for biomedical applications
Lavr Kryukov* and Aleksandr Shirokov Lavr Kryukov Laboratory of Innovative Production Technologies of Plants, Lobachevsky State University, Nizhny Novgorod Region, Russian Federation, Russia *Correspondence: [email protected]
At present, there is almost no reliable data on the biochemical composition of tuberoids. According to the literature, it is only known that they contain starch (2.7%), mucilage (48%), glycosides, polysaccharides, dextrin, minerals, essential oils, proteins, etc. The contents of the components are not exactly specified, and it is impossible to calculate their percentage. However, this information is important, as this plant material could be of great interest to the pharmaceutical and parapharmaceutical industry, because it contains polysaccharides that have biologically active properties. We studied Orchis traunsteineri So3 and Orchis militaris L. from the collection of the botanical garden of UNN. To obtain the tubers, plants were cultivated for 2 months on the artificial agar medium without hormones with 12-hour photoperiod at 18°C and at 60% relative humidity. For the initial material we used protocorms, which were cultured to the stage of the seedling. Root-tubers of orchids contain highly vacuolated aquifer parenchyma cells. We extracted mucoid substance from these cells, dried them, and analyzed them using Shimadzu gas chromatography-mass spectrometer. The analysis showed that the mucilage contains the variety of secondary metabolites, among which we identified sericic acid (inhibition of yeast alpha-glucosidase), lupeol (possesses anti-tumor properties) and betulin (antioxidant and anti-inflammatory properties, immunomodulator). These valuable substances with a strong biological activity were discovered by us for the first time in this group of species. Our further research focuses on the selection of a line of orchid clones containing the largest number of biologically active substances for the biomedical applications.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 60 Day 2: December 12, 2015
Concurrent Session G
AOS networking in seed system biology
Subhash Chandra Naithani*
Seed Biology Laboratory, School of Life Sciences, Pt. Ravishankar Shukla University, Raipur-492010, Chhattisgarh, India S.C. Naithani *Correspondence: [email protected]
Ever expanding but fragmentary literature on the functions of AOS in the critical regulation of various aspects of seeds and seedlings demands comprehensive understanding of AOS networking in a consolidated manner appreciating seed system biology. Dual function of AOS is now established. Depending upon the physiological concentrations of AOS they may play an active role in either the regulation of signal transduction or cellular damage. Our study revealed that the low concentrations of AOS regulate maturation drying of seed development, germination and seedling establishment. Whereas, excess amounts of AOS are central to widely acknowledged “free radical theory” for explaining the mechanism of ageing in several forest tree seeds. Understanding of AOS functions becomes more complicated due to its various biologically active forms such as superoxide, H2O2 and hydroxyl radical with significant variation in their shelf-life and oxidation potential. Acquisition of desiccation tolerance, one of the remarkable traits of seeds to survive vicissitudes of environment is induced by ABA mediated AOS that up regulates Lea and AOS detoxifying genes during maturation drying of seed development of Azadirachta indica. Similarly, AOS signalling coordinate various phases of germination of A. indica seeds by regulating the antioxidant enzymes and cell cycle proteins. In germinated pea seeds, AOS have been assigned a new role in the cell proliferation and differentiation in stem cell niche through controlling the cellular redox state thus important in seedling establishment. Our work is an attempt towards understanding and unifying the tapestry of AOS network in seed system biology.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 61 Day 2: December 12, 2015
Concurrent Session G
Relationship of flag leaf photosynthesis with stomatal density, dry matter partitioning and yield in wheat (Triticum aestivum)
Kushal Kumar Baruah*, Ashmita Bharali and Gulshan Jha K.K. Baruah Department of Environmental Science, Tezpur Central Univ., Napaam-784028, Assam, India *Correspondence: [email protected]
Changing atmospheric levels of carbon dioxide (CO2) and the increasing demands for food of a growing world population are testing the photosynthetic adaptability of a crop and natural ecosystems. A field experiment was conducted during 2013-2014 and 2014-2015 to investigate the efficiency of 07 wheat cultivars [CBW-38(V1), WH-283(V2), WH-1080(V3), WH-1021(V4), WH-711(V5), WH-542(V6) and PBW-343(V7)] for carbon dioxide fixation, dry matter partitioning and yield. Differences among the varieties in terms of their photosynthetic efficiency and biomass partitioning ability were clearly observed. Stomatal frequency of the flag leaf showed a good correlation with photosynthesis at panicle initiation stage. Average number of stomata observed by Scanning Electron Microscopy ranged from 61-111 mm-2 of leaf area of the wheat cultivars. The internal to ambient CO2 mole fraction ratio also differed amongst the varieties which may be attributed to the differences in CO2 requirement of mesophyll cells. Higher stomatal density and flag leaf photosynthesis in the variety WH 1021 contributed to efficient translocation of assimilates to the developing sink and recorded higher dry weight of the kernels at harvest. The pooled analysis of data for both the years of experimentation have revealed that cultivar WH 1021 is superior over other cultivars in terms of photosynthesis and grain yield, and may be beneficial in a future climate change agriculture. Keywords: leaf gas exchange, stomatal density, yield, wheat, cultivar.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 62 Day 2: December 12, 2015
Concurrent Session G
Enhancing stress tolerance by Silicon in rice
S.R. Voleti1*, D. Subrahmanyam1, P.R. Rao1 and A.P. PadmaKumari2
1Plant Physiology Section 2Entomology Section, Indian Institute of Rice Research, Rajendranagar, Hyderabad-500030, Telengana, India S.R. Voleti *Correspondence: [email protected]
The beneficial effects of silicon in plant nutrition particularly abiotic and biotic stress tolerance in rice was studied and reviewed. Lower solubility, uptake, redistribution within the plant organs and polymerization of silicon on the leaf surface are proposed to be important in rice crop. At national level, in collaboration with IICT, compounds solubilizing silica were developed and examined their efficacy under field situation and against various stresses. Enhanced solubility and chemical profile of molecular complexes and sandwich arrangement model was used to explain the mechanism at the soil-plant interface. From yellow stem borer infected plants, wearing larval mandibles in incisor region due to feeding on stems was recorded. Histological evidences suggesting rupturing of peritrophic membrane, vacuolation, disintegration of columnar cells and discharging of cellular contents into the lumen due to abrasion of midgut epithelium using SEM, and deposition of silicon in cell wall in the form of thick dumb-bell shaped bodies using Electron dispersive X-ray spectroscopic analysis of stem samples of rice varieties indicated 11-18% accumulation. Enhanced photosynthesis and water relation characteristics are some of the physiological characteristics that were recorded on silicon application. The beneficial effects studied across the country in rice growing region under AICRIP revealed that, problematic soils are benefitted with application. At International level transporter mediated uptake of soluble silica, fine mapping of genes using low silicon rice mutant, sequencing and identification of aminioacid region and their position are some of the important milestones in this emerging area of silicon research.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 63 Day 3: December 13, 2015
Plenary Session-III
Methylglyoxal and glyoxalase pathway: markers for plant stress tolerance
Charanpreet Kaur1,2, Ananda Mustafiz1, Ajit Ghosh1, Shweta Sharma1, Khirod Sahoo1, 2 1 1,2 Ashwani Pareek , Sneh L. Singla-Pareek and S.K. Sopory * S.K. Sopory
1Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi-110067, India 2School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Correspondence: [email protected]
Methylglyoxal (MG), a potent cytotoxic α-oxoaldehyde is generated as a byproduct of glycolysis and increases to toxic levels under stress and pathological conditions in plants and animals. The ubiquitously present glyoxalase pathway, comprising glyoxalase I (GLYI) and glyoxalase II (GLYII) genes, is required for the detoxification of methylglyoxal. We have shown earlier that, glyoxalase pathway is induced in response to various abiotic stress conditions in plants and over-expression of GLYI and/or GLYII genes impart tolerance towards different stress conditions by keeping MG levels in control, thus indicating their essential role in stress adaptation in plants. Lately, we have shown that the plant kingdom possesses multiple isoforms of GLYI and GLYII which have probably evolved to adopt specialized roles, be it in different organelles, under different developmental stages or in response to different stimuli. A unique Ni2+-dependent GLYI, a nucleus-localized Zn2+-stimulated GLYI and a glutathione-stimulated GLYII have been characterized in rice. Our studies also indicate that MG plays a role in stress signaling in plants, inducing large-scale changes in plant signaling transcriptome and also altering the expression of various genes involved in stress and defense response. Thus, in view of general inducibility of both MG and glyoxalase pathway under stress, these can be proposed as possible biomarkers in plant stress response.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 64 Day 3: December 13, 2015
Plenary Session-III
Insights into plant and algal metabolism through metabolic modelling
Mark Poolman1*, Kailash Adhikari1 and Dipali Singh1
1Department of Biology and Medical Science, Oxford Brookes University, Oxford, U.K. Mark Poolman *Correspondence: [email protected]
In this presentation, some of the fundamental principles of the structural analysis of metabolic networks will be introduced, with particular reference to the application of linear programming to such networks. This will be followed be recent examples of the application of these techniques to the metabolism of Arabidopsis, identifying potential pathways which could circumvent the effect of genetic knockouts which might otherwise be predicted to be lethal, and to the metabolism of the diatom, Phaeodactylum tricornutum, to identify novel potential variants of photorespiration, and roles for enzymes whose purpose is currently unclear. This work is funded as part of the “AccliPhot” project, EU grant number PITN-GA-2012-316427, http://accliphot.eu.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 65 Day 3: December 13, 2015
Concurrent Session H
Discovery and creation of genetic variation to enhance the yield potential of wheat
Martin A.J. Parry* Martin A.J. Parry Lancaster Environment Centre, Lancaster University, Lancaster, UK *Correspondence: [email protected]
There is an urgent need to develop integrated and sustainable approaches that will significantly increase both crop production per unit land area and resource use efficiency. Compelling evidence from free-air CO2 enrichment experiments demonstrates that increasing photosynthesis increases yields, provided that other constraints do not become limiting. The efficiency with which the sun’s energy can be captured as wheat biomass seldom exceeds 2% and averages less than 1%; improving this conversion efficiency is widely recognized as the new frontier for crop improvement. Our research has focused on overcoming the limitations of the CO2-fixing enzyme Rubisco, either through mining natural diversity or by genetic engineering. We have surveyed a large panel of commercial wheat varieties to assess the extent of natural variation in photosynthesis in relation to yield. Significant heritable natural variation in CO2 assimilation was identified. Since photosynthesis is co-limited in the canopy by the kinetics of Rubisco and the regeneration rate of RuBP, increasing RuBP regeneration by overexpressing rate limiting enzymes involved in RuBP regeneration is predicted to increase photosynthesis. We have genetically modified UK and CIMMYT wheat cultivars to increase RuBP regeneration and preliminary results support the predictions of improved photosynthesis at high CO2. Understanding and exploiting natural diversity and creating novel germplasm based upon exploratory research provide a powerful combination to achieve the challenging and pertinent goal.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 66 Day 3: December 13, 2015
Concurrent Session H
Photorespiration: An adaptive response of plants of oxidative stress
Agepati S. Raghavendra*, Bobba Sunil, Vetcha Aswani, Ramesh B. Bapatla and Mohd. Khan Fareed
Department of Plant Sciences, School of Life Sciences, University of Hyderabad, A.S. Raghavendra Hyderabad-500046, India *Correspondence: [email protected], [email protected]
The process of photorespiration achieves two important goals of metabolizing 2-phosphoglycollate, a toxic metabolite, back into the Calvin cycle, and limiting the loss of carbon. The protection by photorespiration of photosynthesis against photoinhibition is well known. We emphasize that photorespiratory reactions contribute to the redox-homeostasis and are part of the adaptive responses of plants to abiotic stress. Photorespiratory reactions can dissipate excess reducing equivalents and energy either directly (by utilizing ATP, NAD(P)H and reduced ferredoxin) or indirectly (by sustaining photosynthetic cyclic electron flow, alternative oxidase (AOX) and providing an internal CO2 pool). Further, photorespiratory H2O2 could be a signal to modulate relevant gene expression. Our concept of photorespiration as an important mechanism to meet the challenge of abiotic stress, is based on multiple evidences. Even mild disturbances in redox status created by deficiency in ascorbate, alternative oxidase or chloroplastic NADP-malate dehydrogenase, cause increases in photorespiratory components, such as catalase, P-protein of glycine decarboxylase complex (GDC) and glycine. In a reversal of roles, any deficiency in photorespiratory capacity, results in oxidative stress. Plants exposed to mild or severe oxidative stress initiate multiple mechanisms to dissipate excess energy and to prevent ROS accumulation. Photorespiration as an important component of such flexible systems, to optimize photosynthesis, and to protect against oxidative stress. Photorespiration integrates the function of chloroplasts, peroxisomes, mitochondria and cytoplasm. Therefore, the strong interaction between the cellular redox status and photorespiration is not surprising, but demonstrates an interesting network of metabolic systems, including photorespiration, to prevent oxidative damage.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 67 Day 3: December 13, 2015
Concurrent Session H
Genetic manipulation of Uroporphyrinogen III methyltransferase (Upm1) modulates nitrogen and carbon assimilation in Arabidopsis thaliana
Sampurna Garai and Baishnab C. Tripathy* B.C. Tripathy
School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Correspondence: [email protected]
The biosynthesis of siroheme, an Fe-containing modified tetrapyrrole, is one of the common links between C and N assimilation. Siroheme, the prosthetic group of nitrite reductase (NiR), is synthesized from uroporphyrinogen III, an intermediate of biosynthetic pathway of chlorophyll and bacteriochlorophyll that are essential for photosynthetic carbon assimilation of eukaryotic and prokaryotic primary producers. The first enzyme in siroheme biosynthesis pathway that diverts uroporphyrinogen III from chlorophyll biosynthesis to siroheme is methyltransferase that catalyzes two methylation reactions of the uroporphyrinogen III to form dihydrosirohydrochlorin that undergoes oxidation to form sirohydrochlorin which is acted upon by sirohydrochlorin ferrochelatase to generate siroheme. The plastidic uroporphyrinogen III methyl transferase coded by a nuclear AtUpm1 [At5g40850] was used to raise transgenic sense and antisense plants to modulate siroheme biosynthesis. Overexpression of AtUpm1 resulted in higher NiR gene and protein expression and enzymatic activities in sense plants than that of WT. It co-modulated nitrate reductase (NR) gene and protein expression, and increased NR activity. Enhanced NiR and NR activities in overexpressors increased their protein content. Higher protein and chlorophyll in sense plants contributed to increased photosynthetic electron transport, carbon assimilation and plant biomass. AtUpm1 overexpression in sense plants protected them from nitrogen deficiency. In contrast antisense plants had lower NiR, NR activities, protein content, carbon assimilation and biomass than the WT and were prone to N starvation. Results demonstrate the co-regulation of two different but inter-dependent biological processes of C and N assimilation by AtUpm1.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 68 Day 3: December 13, 2015
Concurrent Session H
BBX32 controls a network of high light-induced transcription regulators, photosynthesis and dynamic acclimation in mature leaves
Philip Mullineaux1*, Matthews Jack1, Chernukhin Igor1, Jenkins Dafyd2, Wild David2, 1 1 2 1 Bechtold Ulrike , Lawson Tracy , Penfold Christopher , Ruben Alvarez-Fernandez and Philip Mullineaux Beynon James2
1University of Essex, Colchester, UK 2University of Warwick, UK *Correspondence: [email protected]
When low light (LL) - grown plants are exposed to high light (HL) for several days their mature leaves can substantially increase their photosynthetic capacity. This is termed dynamic acclimation and is associated with increased plant productivity. We hypothesised that nuclear-encoded genes coding for transcription regulators (TRs) that respond rapidly and transiently to HL could control the establishment of dynamic acclimation. To find such regulatory genes, a highly replicated temporally clustered transcriptomics dataset was generated from the leaf 7 of Arabidopsis plants exposed for up to 6h HL. The analysis revealed 3844 genes that were responsive to HL. These genes included a significant enrichment of those involved in protection against stress and others that were associated with longer term acclimation. HL-responsive TR genes were selected for Variational Bayesian State Space Modelling (VBSSM), which defined a 15 gene regulatory network (GRN) centred on B-BOX TRANSCRIPTION (CO) FACTOR32 (BBX32). BBX32 was found to be a negative regulator of photosynthesis in LL-grown plants and dynamic acclimation to daily repeated exposure to HL. A major, but not exclusive contribution to BBX32 regulation of dynamic acclimation was through its interaction with LONG HYPOCOTYL5 (HY5), a member of the TR GRN. Our studies indicate that BBX32 is at the centre of a complex temporal regulation of HL-responsive cellular processes underlying its TR GRN.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 69 Day 3: December 13, 2015
Concurrent Session H
Engineering a C4 rice: An update from the international C4 rice consortium
Anindya Bandyopadhyay* Anindya Bandyopadhyay C4 Rice Center, International Rice Research Institute, Los Baños, Laguna, Philippines *Correspondence: [email protected]
The C4 rice project is a large collaborative program involving sixteen separate organizations world-wide (The C4 Consortium), led by IRRI. The aim of the International C4 rice consortium is to produce a large and sustainable increase in the productivity of rice in all ecosystems. To achieve this, the engineering of a C4 photosynthetic pathway into rice (a C3 crop) is underway. C4 cereals typically have grain yields 50% greater than those of C3 crops like rice and wheat. This is because C4 plants have a more efficient photosynthetic pathway that leads to higher photosynthetic rates and improved nitrogen and water use efficiencies. The carbon concentrating mechanism of C4 allows CO2 to be fixed at much lower leaf internal CO2 concentrations. This allows stomatal conductance to be reduced and this lowers the rate of transpiration and doubles water use efficiency. Most of the C4 biochemical pathway genes, along with relevant transporters, have been transgenically introduced into rice so far. These genes are now pyramided and brought into a similar background as a breeding stack. Initial analyses of pyramided lines show signs of a C4 biochemical pathway. Gene discovery for Kranz anatomy and plastid development is also ongoing with interesting transgenic gene candidates under molecular verification. Progress has also been made in the areas such as development of gene stacking technologies for targeted stacking of C4 genes and elucidating the underlying mechanism of stomatal conductance and water use efficiency.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 70 Day 3: December 13, 2015
Concurrent Session H
The dynamic role of cell wall polysaccharides in sustaining sugar homeostasis during leaf senescence of Arabidopsis thaliana: Induction of cell wall hydrolases and cellular sugar reprogramming
Basanti Biswal* Basanti Biswal
CSIR, School of Life Sciences, Sambalpur University, Jyotivihar-768019, Odisha, India *Correspondence: [email protected]
The polysaccharides bound to cell wall are the richest source of organic carbon in plant body and play a critical role in cellular sugar metabolism and homeostasis during plant development and senescence. We have demonstrated that senescence induced decline in photosynthesis in Arabidopsis leaves causes loss of sugars, which acts as a signal for induction of many senescence associated genes (SAGs) that activate several catabolic events to provide sugars from other sources, primarily cellular macromolecules, to complete energy dependent senescence program. Expression of SAGs follows a temporal pattern, some expressed early and some expressed late. These genes initiate and execute an organized and time dependent senescence program. One of the late expressive SAGs codes for bita- glucosidase responsible for breakdown of cell wall polysaccharides. In addition, other wall bound hydrolases including bita-glucanase and bita-galactosidase are activated late during senescence, which suggests that breakdown of cell wall polysaccharides is the terminal event of catabolic network that provides sugars to energy starving cells for completion of senescence program. That the loss of photosynthetic production of sugar as a signal for induction of cell wall bound hydrolases is confirmed with the similar response of the hydrolases during loss of photosynthesis in green leaves of Arabidopsis experiencing abiotic stress. Finally, a model is proposed to explain the nature of signaling involved in cellular sugar reprogramming during plant senescence.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 71 Day 3: December 13, 2015
Concurrent Session H
Photosynthetic efficiencies among Indian peanut cultivars evaluated
A.L. Singh1*, R.N. Nakar2, K. Chakraborty1, Nisha Goswami1 and K.A. Kalariya3
1ICAR-Directorate of Groundnut Research, PB 5, Junagadh-362001, Gujarat, India 2Presently at Department of Botany, Seth PT College, Godhra-389001, Gujarat, India A.L. Singh 3Presently at ICAR-Directorate of Medicinal and Aromatic Plants Research, Boriavi, Anand-387310, Gujarat, India *Correspondence: [email protected]
Peanut is an important legume and oilseed crop of tropical world mainly grown as rainfed in semi-arid regions, and India has the largest area and released cultivars. To know how physiologically efficient our peanut cultivars are?, the photosynthesis (PN), transpiration (E), stomatal conductance (gs), chlorophyll fluorescence, SCMR and yields were studied in field grown 186 cultivars during rainy season where large variability was observed. Addition or deduction of -2 - standard deviation from the average value of these parameters categorized 27 cultivars as high PN with >23.9 μ mol m s 1 -2 -1 -2 - and 30 cultivars as lowPN with <15.7 μ mol m s , 35 cultivars as high E with >8.9 and 24 as low E with <4.7 m mol m s 1 -1 ,35 cultivars as high gs with >0.33 m sec and 28 as low gs with <0.17, 32 cultivars as high WUE >3.81 and 36 as low WUE <2.31. These parameters accounted for the yield potential and 33 cultivars with >225 g m-2 pod were high yielder, -2 -2 26 as low yielder <133 g m pod and 127 cultivars medium with 133-225 g m pod. Positive correlation between, PN and gs, PN and E, PN and number of pods, SCMR and pod yield, SCMR and WUE, haulm yield and plant height, haulm yield and pods, plant height and pods, and a negative correlations between SCMR and plant height, SCMR and E, and E with
WUE were found, however there was no any correlation of PN with yield.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 72 Day 3: December 13, 2015
Concurrent Session I
Male germline development and function in flowering plants
Mohan B. Singh*, Etsuko Itabashi and Prem L. Bhalla
Plant Molecular Biology and Biotechnology Laboratory, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Victoria 3010, Australia Mohan B. Singh *Correspondence: [email protected]
The life cycle of flowering plants alternates between the diploid sporophyte and haploid gametophyte phases, with the latter producing gametes required for sexual reproduction. Male gametophyte development encompasses meiotic division of microspore mother cells giving rise to four haploid microspores. The asymmetric division of the haploid microspore into two daughter cells that produces a small generative cell from the larger vegetative cell initiates the male germline. The generative cell eventually undergoes mitotic division to form two sperm cells required to fuse with two female gametes (egg and central cell) initiating development of the embryo and endosperm, respectively. The molecular basis for the diversification of the cell fates of generative and vegetative cells is a central issue in plant reproductive biology. This diversification is underpinned by the vastly distinct genetic and epigenetic programming of these two pollen cells. The vegetative cell gene expression program emphasizes cell-cell signalling and cell wall organization that might be required for cell-cell communication and pollen tube elongation from pollination to fertilization, whereas the GC transcriptional program is skewed towards DNA/RNA metabolism, histone modification, and cell cycle progression, indicating that gene expression regulation, changes in the epigenetic state and mitotic cell division are associated with the generative cell function. This talk will highlight molecular regulatory pathways controlling male germ line development and function in the flowering plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 73 Day 3: December 13, 2015
Concurrent Session I
Genomic assisted improvement of rice
A.K. Singh*, S. Gopala Krishnan, P.K. Bhowmick, R.K. Ellur, B. Haritha, M. Nagarajan and K.K. Vinod
Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India A.K. Singh *Correspondence: [email protected]
Exemplary grain and cooking quality characteristics along with the pleasant aroma is the uniqueness of Basmati rice. Genetic improvement of Basmati rice has led to the development of varieties with high yield and exceptional cooking quality traits. These varieties have revolutionized the livelihood of the farmers and increased the annual forex earnings to the tune of ~US $ 5 billion through export of Basmati rice. However, Basmati rice was highly susceptible to various biotic stresses such as bacterial blight (BB), blast, sheath blight (ShB), brown plant hopper (BPH) and abiotic stresses such as salinity. Molecular breeding has been successfully utilized in development of several MAS derived varieties such as Improved Pusa Basmati 1 and Pusa 1592 for resistance to BB; Pusa 6 (Pusa1612) and Pusa Basmati 1609 for blast resistance. Further, several monogenic and pyramided NILs with resistance to BB (xa13, Xa21, Xa33, Xa38), blast (Pi54, Piz5, Pi9, Pi1, Pita, Pib, Pi5), sheath blight (qSBR11-1), BPH (Bph18, Bph20 and Bph21), Bakanae (qBK1), salinity stress (Saltol) and herbicide tolerance in the genetic background of popular Basmati rice varieties such as Pusa Basmati 1, Pusa Basmati 1121, Pusa Basmati 6 and Pusa Basmati 1509 are being developed. QTLs for drought tolerance such as qDTY1.1, qDTY2.1, qDTY2.2 and qDTY3.1 are being incorporated into the genetic background of Pusa Basmati 1 and Pusa 44. Furthermore, pro-vitamin A trait has been incorporated into the mega rice variety Swarna.Genomic tools have been of great importance to discover novel QTLs for yield and yield related traits, grain and cooking quality traits, development of heterotic pools as well asassessing genetic purity.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 74 Day 3: December 13, 2015
Concurrent Session I
A novel mutagenesis design to revolutionize genetics, genomics and breeding in maize and beyond
Gurmukh S. Johal1*, Brian Dilkes2, Rajdeep S. Khangura1 and Norman Best3 Gurmukh S. Johal 1Department of Botany and Plant Pathology, Purdue University, West Lafayette, Indiana, USA 2Department of Biochemistry, Purdue University, West Lafayette, Indiana, USA 3Department of Horticulture, and Landscape Architecture, Purdue University, West Lafayette, Indiana, USA *Correspondence: [email protected]
The success of any genetic or breeding program depends on having adequate diversity in the germplasm. This genetic diversity can be of natural origin or generated deliberately by mutagenesis. Various reagents exist to intentionally induce mutations in plants, but alkylating agents such as ethyl methanesulfonate (EMS) are among the most frequently used. Unfortunately, many mutagenesis methods, pollen mutagenesis of maize in particular, are highly inefficient due to a relatively low number of transmitted mutations. As a result, many promising uses of mutagenesis are not feasible or cost effective. We have developed a technique for stacking mutations to increase the mutation density per individual. This increases tremendously the efficiency of independent mutant alleles, thereby opening up multiple applications impossible until now. The process utilizes a multigenerational design to maximize independent changes in the genome and allows screening of subtle differences in mutant phenotypes. In addition, it results in a vast immortalized mutant seed library that can be utilized again and again for multiple independent purposes and benefits. A key benefit is that it maximizes the frequency of polymorphisms of interest for enhancing elite breeding material.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 75 Day 3: December 13, 2015
Concurrent Session I
Ectopic expression of Arabidopsis thaliana HB17 gene in corn leads to improved sink potential
Vijay K. Sharma*, Manju Manjunath, Tom Slewinski and Elena Rice Vijay K. Sharma Monsanto Company, 700 Chesterfield Parkway West, Chesterfield, MO 63017 USA *Correspondence: [email protected]
As a result of the large scale screening of candidate genes in transgenic corn, we identified an Arabidopsis thaliana gene HB17, a member of homeodomain-leucine zipper II (HD-Zip II) family of the plant transcriptional factors, which affects plant growth and leads to increase in ear size at silking. When expressed in corn, AtHB17 lacks the repression domain due to the corn specific splicing mechanism and loses the ability to bind the co-repressors and affect transcription of the target genes. The protein still can form homo-dimers as well as heterodimers with corn endogenous HD-Zip II proteins and bind to the target DNA sequences due to the presence of the functional leucine-zipper and DNA-binding domains. We propose that AtHB17 expressed in corn mediates physiological effects through dominant-negative mechanism by attenuating transcriptional repression activity of endogenous corn HD-Zip II proteins. We hypothesize that modulation of the activity of HD-ZIP II proteins leads to modulation of corn plant’s growth responses to environmental and developmental signals and, ultimately, to increased ear size, thus, providing opportunity for enhanced sink potential in corn plants. Increased sink potential could be manifested through an increase in kernel weight or kernel number depending on the environmental conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 76 Day 3: December 13, 2015
Concurrent Session I
Developing herbicide tolerant rice plants for effective weed management
Aakrati Agarwal, Dhirendra Fartyal, Mrinalini Manna, Mohan Murali Achary, Vijay Sheri, Babu Ram Verma, Tanushri Kaul and M.K. Reddy* M.K. Reddy
International Centre for Genetic Engineering and Biotechnology, New Delhi-110067, India *Correspondence: [email protected]
Weeds are the most serious biological constraint in agriculture and manual weeding over large areas is not feasible. In this scenario, introduction of genetically engineered herbicide resistant crop plants will be a better weed management strategy. We developed transgenic rice plants simultaneously resistant to two broad-spectrum systemic herbicides i.e., sulfonylurea for pre-emergent and glyphosate for post-emergent application. Sulfonylurea is a very strong inhibitor of acetolactate synthase and glyphosate is a strong inhibitor of EPSP synthase the important plant enzymes involved in amino acid synthesis. We followed herbicide target site modification strategy by introducing selective amino acid substitutions in rice EPSP synthase and acetolactate synthase. These multisite-compensating mutations drastically decreased affinity for herbicide binding without detectable fitness cost. These mutant copies of rice EPSP synthase and acetolactate synthase encoding genes were cloned under constitutive promoters and linked together on a single plant transformation vector for simultaneously transformation into rice. These transgenic rice lines showed significant tolerance to glyphosate and sulfonylurea. Systematic rotation of glyphosate and sulfonylurea application in the rice fields is expected to prevent the development of herbicide tolerant weeds. We have also developed another strategy to control the weeds by application of phosphite. Plants cannot metabolize phosphite, as they can only utilize phosphate for their growth. We have developed transgenic rice plants overexpressing phosphite dehydrogenase that converts phosphite into phosphate. Phosphite can act as a fertilizer for the transgenic plants whereas, for weeds it acts like a herbicide and act as herbicide for the weeds.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 77 Day 3: December 13, 2015
Concurrent Session I
Identification and functional characterization of the regulators of reproductive development in rice
Kapoor Sanjay*, Chawla Mrinalini and Nayar Saraswati Sanjay Kapoor Department of Plant Molecular Biology, Univ. of Delhi South Campus, New Delhi-110021, India *Correspondence: [email protected]
Reproductive development in plants is a dynamic process that involves complex interplay of regulatory networks that are under the influence of genetic as well as extrinsic cues. Our group has been involved in the identification and characterization of both genic and epigenetic components that affect development of the pollen and the seed. Our studies on the rice model have been instrumental in the identification of sets of genes that could potentially play key roles during sporophytic to gametophytic transition in pollen mother cells, pollen maturation, embryo differentiation and grain filling. Male gametophyte development has been shown to be a heat-sensitive process in a variety of plant species and is one of the major determinants of yield in many crops. We have identified components of RNAi and ubiquitin-mediated protein degradation machinery that mimic pollen heat phenotypes, with reduced starch accumulation and loss of viability. We have also identified and carried out functional delineation a seed-specific MADS box transcription factor, OsMADS29, that regulates embryo development and grain filling by affecting hormone homeostasis during seed development in rice. Rice MADS29 has also been found to interact with several seed-expressed transcription factors and its capability to localize in the nucleus is affected by these interactions. These and other leads could thus contribute genetic targets and tools for augmenting yield as well as nutritive value of the seeds.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 78 Day 3: December 13, 2015
Concurrent Session I
Towards integrated product development
Amit Das1, Sresty Tavva1, Ratna Kumria1, Priyanka Bhyri1, Bindu Andreuzza1, Hajime Sakai2, Thomas W Greene3 and Amitabh Mohanty1*
1DuPont Pioneer, DuPont Knowledge Centre, Turkapally, Hyderabad-500078, Telangana, India Amitabh Mohanty 2E.I. duPont Nemours and Company, DuPont Experimental Station, Wilmington, Delaware 19898, USA 3DuPont Pioneer, 7000 NW 62nd Ave, Johnson, IA, USA *Correspondence: [email protected]
The global population is estimated to reach 9 billion by the year 2050. Most of this population growth is occurring in the region of low agricultural productivity. With the ever increasing population, there is a need to produce more food to feed the growing population. Since agricultural productivity has not kept in pace with the population increase and therefore, there is a strong need to use cutting edge technology for improving agricultural productivity, especially for important traits like nitrogen use efficiency (NUE) and drought tolerance (DRT). We have employed forward genetics, reverse genetics, as well as new breeding technologies, such as CRISPR/Cas9-based guided genome editing to address some of the key issues. Recent advances in development of tools for precision genome engineering, such as site directed nucleases (SDN) have opened up a plethora of opportunities for plant breeding and genetics. These new breeding technologies (NBTs) allow us to perform targeted and precise modification of the genome, thereby imparting ability to design and develop a new generation of crops with improved traits. Application of these new breeding technologies has potential to result in significant gains in the area of crop improvement. In this presentation we will present a holistic view of our integrated approach towards product development.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 79 Day 3: December 13, 2015
Concurrent Session J
Unraveling key regulators of floral transition at the shoot apical meristem of soybean
Prem L. Bhalla*, Chui E. Wong, Chol-Hee Jung, Lim Chee Liew and Mohan B. Singh Prem L. Bhalla Plant Molecular Biology and Biotechnology Laboratory, Faculty of Veterinary and Agriculture Sciences, The University of Melbourne, Parkville, Victoria 3010, Australia *Correspondence: [email protected]
Flowering process controls crop productivity. Understanding of the floral transition is essential in ensuring future food security. Soybean [Glycine max (L.) Merr.], one of the world’s most important crops, is responsible for 55% of the worldwide oilseed production and a significant source of dietary proteins. Further, soybean is significant in the world agriculture due to its capacity to fix atmospheric nitrogen. Soybean is a photoperiod sensitive crop whose floral transition is triggered by exposure to short-day conditions. Our knowledge of the molecular control of flower initiation in this economically important legume species is limited. We used an integrated experimental and bioinformatic approach to addresses this gap in our knowledge. Our comparative genomics study revealed complete repertoire of flowering regulatory genes in the soybean genome. Further, RNA-Seq analysis of shoot apical meristem and leaf undergoing floral transition revealed major reprogramming events in leaf and the shoot apical meristem that point toward hormones gibberellins and cytokinin as key regulators in systemic flowering signal(s) production in leaves. Our data also revealed an extensive reprogramming of genes associated with the epigenetic chromatin modifications and RNAi gene silencing in the shoot apical meristem during floral transition. Our results highlight the use of a genomics-based approach in improving our understanding of this important developmental process and provided a framework to investigate further the molecular network underlying the transition to flowering in soybean.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 80 Day 3: December 13, 2015
Concurrent Session J
Vernalization and epigenetic regulation in Arabidopsis
Christopher Helliwell1*, Masumi Robertson1, Jean Finnegan1 and Bob Anderssen2
1CSIRO Agriculture, GPO Box 1600, Canberra, ACT 2601, Australia 2Data61, GPO Box 664, Canberra, ACT 2601, Australia Christopher Helliwell *Correspondence: [email protected]
Many plants, including Arabidopsis, respond to extended periods of cold (vernalisation) by initiating the transition from vegetative to reproductive development. We are interested in this process both because of its biological importance and as a model for gene regulation by chromatin modification. In Arabidopsis the vernalisation response is conferred by expression of the FLC gene, a repressor of the flowering pathway. FLC transcription is down regulated in response to cold and this down-regulation is maintained once plants returned to warm conditions. The maintenance of FLC repression requires deposition of the chromatin modification H3K27me3 by the polycomb system. I will discuss our understanding of how FLC is repressed by cold, how this repression is maintained and the wider implications of this for understanding the role of chromatin modifications in plant development.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 81 Day 3: December 13, 2015
Concurrent Session J
Physiological and biochemical basis of seed longevity during storage
M.B. Chetti1* and M.K. Meena2
1Assistant Director General (HRD), Education Division, KAB-II, ICAR, New Delhi-110012, India 2Assistant Professor, Department of Crop Physiology, UAS, Raichur-584104, Karnataka, India M.B. Chetti *Correspondence: [email protected]
Great strides in seed packaging and storage have been made under the quality maintenance strategy by using vapour proof packaging materials which in turn made possible the global distribution of high value and long term availability of seeds. The factors affecting the storage life of seeds have been identified and characterized for successful seed storage and accordingly packaging materials are modified considering the storage period. Results of our study revealed that there was a gradual decline in seed quality parameters during entire storage period of 18 months in cotton, groundnut, soybean and onion. Not much variation was observed with respect to the vacuum packed bags from zero to 18 months of storage, irrespective of seeds stored either under cold storage or at room temperature. The changes in seed quality (germination per cent, mean daily germination, root length, shoot length, total seedling length, seedling dry weight, mobilization efficiency and vigour index) were attributed to the changes in moisture content. Comparing the HDPE and gunny treatments under room temperature and cold storage, lower values were observed in the latter, which is attributed to the high moisture in seeds as the cold storage maintains high humidity. The electrical conductivity was significantly lower in seeds stored in vacuum sealed containers followed by polythene bags and was higher in gunny bags and cloth bags throughout the storage period. Higher electrical conductivity values recorded in seeds stored in gunny bags and cloth may be due to higher level of seed deterioration on account of age induced membrane damage of various cell and cell organelles or degradation or disruption of cell membranes leading to subsequent loss of membrane integrity. Our investigations revealed that α-amylase enzyme decreased with an increased storage period. High density polythene, gunny and cloth bags stored seeds recorded higher α-amylase activity compared to vacuum packed seeds followed by cloth bags. Decreased α-amylase activity with an increase in storage period may be due to reduction of free sugars and amino acids. Lipase and protease activity were found to be slightly less in vacuum packed bags compared to aluminum, HDPE, gunny and cloth bags. There was higher lipase activity in gunny bags followed by cloth bags. There was increase in the activity of these enzymes with advancement in storage period. The investigation has thrown light on several aspects that need to be worked out in detail in future.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 82 Day 3: December 13, 2015
Concurrent Session J
How and why of flower senescence – Understanding from models to ornamentals
Inayatullah Tahir* and Syed Sabhi Ahmad Inayatullah Tahir Plant Physiology and Biochemistry Research Laboratory, Department of Botany, University of Kashmir, Srinagar-190006, India *Correspondence: [email protected]
Flower senescence represents the last stage of flower development ultimately resulting in the death of the floral parts. It is regulated by an array of external and internal cues coordinated at the gene level which bear the hallmark of programmed cell death (PCD). Petals provide an excellent model system for deciphering senescence as they have a finite lifespan regulated by tight developmental control. While flowers serve an essential role in sexual reproduction, the maintenance of petals is costly in terms of energy and water loss. PCD in flowers is generally triggered, once pollination takes place and the plant doesn’t require the floral organs. This results in the breakdown of macromolecules/ organelles and remobilization of their products to developing tissues. Flower senescence is chiefly regulated by a complex interplay between various growth regulators like ethylene, abscissic acid, auxins, gibberellins, cytokinins, jasmonic acid, brassinosteroids and polyamines. Ethylene plays main role in some flowers while as in others, it has a little or no role in the execution of flower senescence. Proteolytic cleavage is thought to play a pivotal role in the regulation of senescence in some flowers, being one of the earliest signs of gene change resulting in raised enzyme activity and decline in the protein levels. These changes cause membrane fluidity, increased lipid peroxidation and decreased activity of antioxidant enzymes, ultimately leading to flower death. Our study tries to gain an insight into the phenomenon of flower senescence by looking from models to ornamentals.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 83 Day 3: December 13, 2015
Concurrent Session J
Metabolic dominance of spikelets in rice panicle is determined by genetic expression of starch synthesizing enzymes
Binay Panda and Pravat Kumar Mohapatra* P.K. Mohapatra School of Life Sciences, Sambalpur University, Sambalpur-768019, Odisha, India *Correspondence: [email protected]
Spikelets are the individual flower units of the complex racemose type panicle of rice. Each spikelet produces one seeded fruit caryopsis at maturity; the success of occurrence depends on completion of starch synthesis in the endosperm. The spikelets located on the apical part of panicle dominate metabolically and produce well-filled grains in contrast to the inferior basal spikelets producing poorly-filled grains unsuitable for human consumption. Starch comprises 80% of the final dry weight of rice grain. Poor sink activity in the inferior spikelets underscores genetic potential for starch synthesis to the detriment of grain filling which downgrades grain weight and quality. In the recent years, rice breeders have increased spikelet number of panicle without increment of size resulting in a change over of spikelet arrangement from lax to compact. Compact arrangement compromised with apical dominance and shortage of space between spikelets discouraged starch filling of the extra-spikelets accommodated. The discrimination in starch filling capacity of the inferior spikelets have been measured by estimation of activity of three major starch synthesizing enzymes, such as, sucrose synthase, AGPase and starch synthase and their expression as seen in RT- PCR and qRT-PCR studies. Genetic expression of various isoforms of the enzymes was higher in the lax- compared to compact-panicle rice cultivars and apical verses the basal spikelet. The metabolic dominance of a rice spikelet depends on the expression of the genes for different isoforms of starch synthesizing enzymes, but the expression of isoform three of sucrose synthase could be more specific than the others.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 84 Day 3: December 13, 2015
Concurrent Session J
Understanding environmental and physiological basis for low productivity of Alphonso mango under coastal rainfed ecosystem of Konkan region in India
Murad M. Burondkar*, Sanjay Bhave, Mahesh Kulkarni and Arun Mane Murad M. Burondkar
Dr. B.S. Konkan Krishi Vidyapeeth, Dapoli, Ratnagiri, Maharashtra, India *Correspondence: [email protected]
Konkan region on the west coast of India, is one of the biggest (0.18 million ha) mango (cv. Alphonso), growing belts in India. However, inspite of several technological interventions, very low mango orchard efficiency (2.5-3.0 t/ha), is considered as major long standing constraint. Hence, present study was aimed at establishing the association between the climatic factors prevailing during 3 different seasons (monsoon, winter and summer) and physiological attributes such xylem sap flow, leaf water potential and rate of photosynthesis, recorded fortnightly during on 30 years old Alphonso mango trees, grown on red lateritic soils. Results indicated very close association between weather parameters and physiological response of mango during all the three seasons. Rate of photosynthesis was maximum (8.91 μmol -2 -1 CO2 m s ) only during October to November and was positively favoured by available bright sun light above light saturation point (approx. 500 μmol m-2s-1), maximum xylem sap flow and availability of leaf moisture content for the corresponding period. The rate of photosynthesis during monsoon (June-September) was bare minimum, ranging -2 -1 -2 -1 from 2.0-2.5 μmol CO2 m s , due to very poor sunlight (35-40 μmol CO2 m s i.e. almost ten times below light saturation point. After November, the rate of photosynthesis gradually decreased towards summer on account of lower xylem sap flow and leaf water potential (-1.9 bar) due to soil moisture stress and low humidity.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 85 Day 4: December 14, 2015
Plenary Session-IV
Investigations on cold acclimation induced changes in photosynthesis in Arabidopsis: Chlorophyll a fluorescence emission measurements
Kumud B. Mishra1 and Govindjee2* Govindjee 1Global Change Research Centre, Academy of Sciences of the Czech Republic, v. v. i, Belidla 986/ 4a, 603 00, Brno, Czech Republic 2Department of Plant Biology, Department of Biochemistry and Center for Biophysics & Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, IL USA *Correspondence: [email protected]
In spite of extensive research on diverse plant species displaying a broad range of cold tolerance, the strategy and dynamic processes involving the onset of cold acclimation induced cold tolerance and regulatory steps controlling re- adjustment of physiological homeostasis at low temperatures are inadequately studied and remain an area for future research. Further, most of the studies on cold tolerance have been carried out on plants grown in static environment, e.g., inside a growth chamber. In nature, plants are adapted to dynamic environment with varying light and temperature throughout their growth cycle, and this dynamic environment must have an important role on the development of tolerance of plants to these environmental stimuli including low temperatures. Since chlorophyll (Chl) a fluorescence emission has already been established as a powerful, and non-invasive tool for measuring various steps in photosynthesis, we have used it in this research to study cold acclimation induced changes in the model plant Arabidopsis (A.) thaliana. We shall present and discuss our results on Chl a emission and relate them to the modulation of photosynthesis in leaves from non-acclimated and cold acclimated natural accession of A. thaliana C24 (native of Pourtgal), during linear cooling from room temperature (220C) to -1.5°C.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 86 Day 4: December 14, 2015
Plenary Session-IV
Ionome to genome: Gene discovery in aid of plant nutrition, human health and environmental remediation
Mary Lou Guerinot* Mary Lou Guerinot Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire, USA *Correspondence: [email protected]
Micronutrient deficiencies commonly limit plant growth and crop yields. If the mechanisms of uptake, distribution and regulation of micronutrients were clearly understood, it might be feasible to engineer plants better able to grow in soils now considered marginal and to increase crop biomass on soils now in cultivation. Furthermore, as most people rely on plants as their dietary source of micronutrients, planlits that serve as better sources of essential nutrients would improve human health. We combine genetics, high throughput elemental analysis via ICP-MS and high resolution imaging via synchrotron X-ray fluorescence to identify and characterize genes involved in metal uptake, distribution and storage. Most of our work has been focused on the essential micronutrient iron. We are characterizing several Arabidopsis mutants that have the potential to improve seed iron content. One of these, Ig14, has a similar metal content to wild type when grown on normal soil, but thrives on alkaline soil, accumulating significantly more iron in its shoot and seeds. Another mutant, 93699, accumulates significantly more iron than wild type and is very sensitive to exogenous iron supply. The mutation in a third mutant has been mapped to an essential bHLH gene we call URI that controls many of the iron-regulated genes in Arabidopsis. We are also taking similar approaches to determine how arsenic, a non-threshold, Class 1 human carcinogen, accumulates in rice, a staple food for over half the world’s population. We have been screening a large collection of rice cultivars and have identified several QTLs for arsenic.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 87 Day 4: December 14, 2015
Plenary Session-IV
Capitalizing on genetic variation for functional genomics of quantitative traits in rice
T. Mohapatra* T. Mohapatra ICAR-Indian Agricultural Research Institute, New Delhi-110012, India *Correspondence: [email protected]
Genetic variaion is essential for evolution. It is the key to the development of improved cultivars through selection of the best. Besides, genetic analysis of any trait requires variation. Variation in natural populations of cultivated and even wild forms are first utilized for the purpose. Induced genetic variation often supplements the natural genetic variation while dissecting target traits. Majority of the traits of agricultural importance are quantitative in nature. Understanding variation in such traits requires different approaches in comparison to Mendelain traits. In recent years, a combination of genetic, genomic and statistical tools/approches is being applied to make sense of the variation in complex traits and to infer on how the genome functions. Rice is the staple food for more than half of the world’s population, especially those living in Asia. However, the world population is increasing at a fast pace, which might reach nine billion mark by 2050. The population of India is also estimated to be around 1.6 billion by that time. To feed this huge population, it will be required to increase production and productivity of rice. This is going to be highly challenging in view of the changing climate that would accentuate the problems due to abiotic and abiotic stresses. As the economic condition improves along with rising purchasing power, there will be greater demand for better quality of the produce. Fortunately, rich genetic diversity exists in rice. More than 100,000 germplasm lines including wild relatives have been collected and conserved in the National Gene Bank of the NBPGR, New Delhi. Subsets of this germplasm have been evaluated for a range of traits including, yield, tolerance to abiotic and biotic stresses and nutritional quality at different institutions in the country. Besides, more than 20,000 EMS mutagenised lines of the upland cultivar Nagina22 have also been developed through national network funded by the Department of Biotechnology, Govt. of India. Laboratory and field evaluation has led to identification of mutants with novel phenotypes. We have made use of these genetic resources to understand rice genome function. The traits that we have targeted include yield components, drought tolerance, salinity tolerance, cold tolerance, anaerobic germination and grain iron content. Whole genome re-sequencing of contrasting genotypes has revealed huge DNA sequence level variation with genome scale distribution. Transcriptome sequencing has led to identification of genes showing differential expression in contrasting lines suggesting thereby involvement of different pathways. Inclusion of EMS mutants in the study has helped identification of newer alleles of known genes. Integration of genome-wide approaches has provided opportunity to understand variation in complex traits such as tolerance to drought and salinity. The results of the above studies will be discussed.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 88 Day 4: December 14, 2015
Concurrent Session K
A day and a night in the life of a unicellular diazotrophic cyanobacterium
Himadri B. Pakrasi*
Laboratory of Himadri Pakrasi, Washington University, International Center for Advanced Renewable Energy and Sustainability, St. Louis, MO 63130, USA Himadri B. Pakrasi *Correspondence: [email protected]
Cyanobacteria are unique among prokaryotes for their ability to perform oxygenic photosynthesis and for their circadian lifestyle. Unicellular cyanobacteria like Cyanothece are fascinating organisms that actively use both phases of the diurnal cycle to separate and carry out antagonistic metabolic processes. The intracellular environment oscillates between aerobic and anaerobic states during the day-night period, allowing oxygen-sensitive processes like nitrogen fixation to occur at night while photosynthetic oxygen evolution takes place during the day. This diurnal periodicity gives a highly dynamic profile to these organisms, which can be captured at the genome, transcriptome, proteome and ultrastructural levels. Whole genome analysis of six Cyanothece strains revealed that this group of unicellular nitrogen fixing cyanobacteria specialize in metabolic compartmentalization and energy storage, concomitantly accumulating metabolic products in inclusion bodies that are later mobilized as part of a robust diurnal cycle. Transcriptomic analyses showed that the existence of these incompatible processes in the same single cell depend on tightly synchronized expression programs involving ~30% of genes in the genome, a phenomenon that also extends to a large extent at the protein level. To examine the proteome of Cyanothece, we used a high-throughput accurate mass and time (AMT) tag approach and identified a total of 3,616 proteins with high confidence, which is approximately 68% of the predicted proteins based on the sequenced genome. We have also examined the dynamic profiles of all of these proteins during a diurnal period. Three-dimensional tomographic reconstructions showed that the thylakoid membranes in Cyanothece form a dense and complex network that extends throughout the entire cell. In particular, the organization of these membranes and various intracellular bodies profoundly change between day and night periods. Together, these studies have provided a comprehensive picture of how a physiologically relevant diurnal light-dark cycle influences the metabolic behavior of a diazotrophic photosynthetic bacterium.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 89 Day 4: December 14, 2015
Concurrent Session K
Accelerating product pipeline through computational biology: deciphering role of Arabidopsis HB17 in maize
Saritha V. Kuriakose1*, Naveen Silvester1, Anuradha Mahindra1, Kyle Skottke2, 2 2 Elizabeth Gohara and Abha Khandelwal Saritha V. Kuriakose
1Monsanto Research Centre, Vasanth’s Business Park, Bellary Rd, Hebbal, Bangalore-560092, Karnataka, India 2Monsanto Company, 700 Chesterfield Parkway West, Chesterfield, MO 63017, USA *Correspondence: [email protected]
As a result of the large scale screening of candidate genes in transgenic corn, we identified an Arabidopsis thaliana gene HB17, a member of homeodomain-leucine zipper II (HD-Zip II) family of the plant transcriptional factors, which affects plant growth. Transformation of maize with an ATHB17 expression construct leads to the expression of ATHB17Δ113, a truncated protein lacking the first 113 amino acids which encodes a significant portion of the repression domain. Because ATHB17Δ113 lacks the repression domain, the protein cannot directly affect the transcription of its target genes. ATHB17Δ113 can homodimerize, form heterodimers with maize endogenous HD-Zip II proteins, and bind to target DNA sequences; thus, ATHB17Δ113 may interfere with HD-Zip II mediated transcriptional activity via a dominant negative mechanism. We used computational approaches to provide first level evidence that maize HD-Zip II proteins function as transcriptional repressors and that ATHB17Ä113 relieves this HD-Zip II mediated transcriptional repression activity. This was later confirmed in targeted experiments. Expression of ATHB17Δ113 in maize leads to increased ear size at silking and, therefore, may enhance sink potential. We hypothesize that this phenotype could be a result of modulation of endogenous HD-Zip II pathways in maize.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 90 Day 4: December 14, 2015
Concurrent Session K
CCPM: a web platform for the data capture, analysis and integration of plant metabolomics
Indira Ghosh1*, Abhijit Mitra2, Vikram Pudi2 and Nita Parekh2 Indira Ghosh 1School of Computational and Integrative Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Center for Computational Natural Sciences and Bioinformatics, International Institute of Information Technology, Hyderabad-500032, India *Correspondence: [email protected]
Plant metabolomics represents a global quantitative assessment of metabolites within a cell sample in specific plant tissue, assisted by advances in high-throughput instrumentation has emerged as a powerful source for the high throughput phytochemical analysis of plants and rapid generation of large volumes of high quality data, signifying different chemicals, in quantity & quality. A collaborative platform has been developed to addresses the unique interdisciplinary character of metabolomics; an integrated and effective tool, for web-based data repository with analysis & visualization platform for plant metabolomics known as CCPM 3.4 (Computational Core of Plant Metabolomics). This is a fully functional, state of art web application, addressing the issues of storage, processing, analysis and sharing of data and results of metabolomics experiments. With a python web framework (web2py) backend and open source analysis programs embedded, CCPM allows rapid prototyping, easy maintenance and role-based access control without compromising on security, scalability and efficiency through a user friendly bootstrap-based GUI. It provides the pipeline for end-to-end analysis of LC/GC-MS data involving raw data capture, data pre-processing, putative metabolite characterization (linking to METLIN and GOLM), and a wide range of statistical analysis, with option for customization of parameters from the web interface, modular architecture allows input of externally preprocessed data in all popular formats, directly to avail the analysis tools. It creates metadata on the details of sample, experimental conditions and factors. Other features include user friendly tools for on the fly data analysis and publication quality image download for experimental metabolomics of plant data.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 91 Day 4: December 14, 2015
Concurrent Session K
Wheat phenomics through hyper-spectral remote sensing for water deficit stress
R.N. Sahoo1*, Bappa Das1, Gopal Krishna1, Rahul Tiwari2, Rakesh Kumar2 and 2 C. Viswanathan R.N. Sahoo 1Division of Agricultural Physics, 2Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India
*Correspondence: [email protected]
Potential of hyper-spectral remote sensing has been explored for non-invasive assessment of water stress for wheat crop. Hyper-spectral data (350 to 2500 nm) and relative water content (RWC) of 10 different genotypes of wheat crop were measured at different stress level. Comparative evaluation of newly developed spectral index with existing water sensitive spectral indices and band depth analysis at selected wavelengths was done with respect to leaf water status, and spectral based prediction models were developed. Then, the proposed ratio of spectral indices (RSI) and normalized difference spectral indices (NDSI) based on the leaf reflectance spectra were obtained in all possible combinations within 350–2500 nm, and their correlation with RWC were quantified to identify the best indices. The best spectral indices for estimating RWC in wheat were found to be, proposed RSI (R1391, R1830) and NDSI (R1391, R1830) with R2 of 0.86 and 0.81, respectively. Hyper-spectral canopy reflectance (350–2500 nm) data recorded at various stress levels were used to develop a partial least squares regression (PLS) model followed by multiple linear regression (MLR) to calculate plant RWC. The model R2 and RMSE was found to be 0.96 and 0.88 and 3.88% and 7.16% for calibration, and 0.91 to 0.88 and 6.52% to 7.54% for validation, respectively. Based on differential spectral response of genotypes at different water deficit stress levels, genotypes were classified into stress tolerant and susceptible types. The methodology developed would be further used in high throughput phenomics of different crops for drought stress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 92 Day 4: December 14, 2015
Concurrent Session K
Mechanistic investigation of floral transition in plants using computational systems biology
Gitanjali Yadav* Gitanjali Yadav Computational Biology Laboratory, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Correspondence: [email protected]
Floral transition represents one of the most critical events in the life of a plant, and refers to the progression from vegetative to reproductive growth. It is known to be determined not by a single gene but by a highly complex gene network, in which LEAFY (LFY), a transcription factor with protein binding function that expresses widely in both vegetative and reproductive tissues, plays an important role as a switch that triggers flower formation by interacting and coordinating between several other genes. We have investigated the floral transition event through a graph- theoretical approach at a genome-wide network level in Arabidopsis thaliana by superimposing mRNA expression profiles and gene ontologies of the known floral genes with the available floral interactome. The regulatory network provides insights into how LFY, as a master regulator, positively or negatively synchronizes the level or activities of other genes in the general physiology of flowering. A dynamic analysis of network topology led to the identification of key regulatory modules of genes during flowering, apart from characterization of relationships among genes in different floral modules. Since LFY stands at the very center of the network of flower development, even minute modifications in its expression/interactions may contribute to the appearance of floral structures in evolution. In summary, this work massively expands the known floral network of Arabidopsis thaliana besides offering new perceptions regarding the principal mechanism of flowering and molecular basis of diverse floral morphologies.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 93 Day 4: December 14, 2015
Concurrent Session K
Comparative analysis of glutaredoxins (GRXs) in monocot and dicot species
Hemant Ritturaj Kushwaha1*, Minesh Jethva1 and Saurabh Yadav2 Hemant R. Kushwaha 1Synthetic Biology and Biofuels Group, International Centre for Genetic Engineering and Biotechnology, New Delhi-110067, India 2Department of Biotechnology, Hemwati Nandan Bahuguna Garhwal University, Srinagar Garhwal, Uttarakhand-246174, India *Correspondence: [email protected]
Environmental variations often lead to the production of reactive oxidative species (ROS), thus causing oxidative stress in plants. One of the enzymatic antioxidant systems, Glutaredoxins (GRXs) has been characterized for its role in redox homeostasis. GRXs are the small thermo-stable thiol based oxidoreductases of thioredoxin (TRX) superfamily. GRXs are involved in the maintenance of cellular redox homeostasis in bacteria, yeast and plant and mammalian cells. Glutaredoxin proteins perform the glutathionylation and de-glutathionylation reactions. Broadly they have been classified into two types based on the redox active motif: Monothiols (CGFS) and dithiols (CXXC). CC-type GRXs are specific to land plants and participates in the more complex functions in the plants. GRXs are widely conserved in both prokaryotes and eukaryotes and are known to have many roles in addition to oxidative stress. In this study, we have identified and characterized GRXs in nineteen monocot and dicot species. We have analyzed domain architecture in their protein sequences, in order to establish their evolutionary relationship. Further, using phylogenetic tree, we have analyzed the conserved features in monothiol and dithiol sequences in both monocots and dicots. The localization of GRXs on various chromosomes of monocots and dicots highlights their probable expansion in the various plant genomes. Thus, this analysis provides novel insights to the functions of GRX proteins in various monocots and dicot species.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 94 Day 4: December 14, 2015
Concurrent Session L
Role of transpiration in regulating leaf temperature in wheat: Physiological basis of contrasting genotypic responses
S.R.W.M.C.J.K. Ranawana1,2,5, H, Bramley4, J.A. Palta1,3 and K.H.M. Siddique2* K.H.M. Siddique 1School of Plant Biology, The University of Western Australia, Perth, WA 6009, Australia 2The UWA Institute of Agriculture, The University of Western Australia, Perth, WA 6009, Australia 3CSIRO Agriculture, Private Bag No. 5, Wembley, WA 6913, Australia 4Plant Breeding Institute, Faculty of Agriculture and Environment, The University of Sydney, 12656 Newell Highway, Narrabri NSW 2390, Australia 5Department of Export Agriculture, Faculty of Animal Science and Export Agriculture, Uva Wellassa University, Badulla 90000, Sri Lanka *Correspondence: [email protected]
The projected increase in global temperature may drastically reduce wheat yields without adaptation strategies. The role of transpiration in dissipating heat in wheat leaves through evaporative cooling was assessed in relation to changes in vapour pressure deficit (VPD), air temperature and soil water under controlled environmental conditions. A reductionist approach was taken starting with evaluating a morpho-physiologically diverse group of 20 wheat genotypes under two temperature regimes for total water use, leaf temperature and biomass production. Selected contrasting genotypes were then evaluated under controlled VPD conditions with varying levels of soil water to identify the physiological basis of the contrasting responses. Greater water use and its effects on leaf cooling contributed to greater biomass production under high temperature, which depended on genotype. Genotypes differed in their leaf temperature regulation that did not always correlate with transpiration response to VPD, indicating other heat dissipation mechanisms are involved in leaf cooling in addition to transpiration. Two contrasting transpiration responses to VPD were identified: (1) Transpiration increased gradually at slower rate with VPD while stomatal conductance was unresponsive, and (2) transpiration responded to VPD in two phases separated by a breakpoint (segmented linear response) where transpiration responds little to VPD after the breakpoint. Genotypic variation in transpiration to VPD response could be used to develop new wheat cultivars targeting specific environments. The first response pattern may be better suited to rain-fed agricultural systems, where yield depends on stored water. The second pattern may be better suited for agricultural systems with intermittent in-season rainfall.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 95 Day 4: December 14, 2015
Concurrent Session L
Functional mechanisms and strategies for improving drought tolerance in rainfed crops
M. Maheswari* M. Maheswari Division of Crop Sciences, Central Research Institute for Dryland Agriculture, Santoshnagar, Saidabad PO, Hyderabad-500059, India *Correspondence: [email protected]
Developing tolerant genotypes is crucial for stabilizing crop yields under drought stress conditions as it is one of the most important abiotic stresses affecting crop productivity. Maize and pigeon pea genotypes were evaluated for water deficit toler-ance across seasons under well watered and water stressed conditions to identify interactions amongst various tolerance traits and grain yield as well as their association with SSR markers. Significant positive correlations were elucidated amongst the water relations traits with yield as well as with a few SSR markers under water stressed conditions indicating the importance of functional mechanisms of intrinsic tolerance and cumulative traits for drought tolerance in rainfed maize. The importance of recovery patterns in identifying tolerant genotypes was highlighted using high through put phenomics. In pigeon pea genotypes, water deficit impacted biomass accumulation, partitioning while it enhanced the flower drop and decreased flower to pod conversion and molecular genetic diversity based on SSR markers associated with drought tolerance revealed useful information for genetic enhancement of drought tolerance. The cross talk of different abiotic stresses is mediated through anti-oxidative metabolism and differential expression of AOX isozymes seemed to be involved in stress tolerance in rainfed crops like pearl millet. Similarly differential expression especially of transcription factors and regulatory genes is a key response in crop plants under climatic stresses. The differential expression analysis of SnRK2, a plant specific protein kinase family involved in stress signalling indicated that this family of genes has a functional role in stress tolerance. Integrated approaches utilizing the phenomics, functional genomics and molecular breeding are essential for enhancing climate resilience in rainfed crops.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 96 Day 4: December 14, 2015
Concurrent Session L
Climate information, crop modeling and impacts on global food security
Vangimalla R. Reddy* and Shardendu K. Singh
Research Leader, USDA-ARS Crop Systems and Global Change Laboratory, Beltsville, MD, USA Vangimalla R. Reddy *Correspondence: [email protected]
Climate change is already affecting the natural resources such as terrestrial vegetation, animal husbandry, and fisheries that societies depend on to provide food, fiber, fuel, several industrial products, and recreational services. These carbon-based products and fresh water supply come from the thin living skin covering the earth’s land surface called the terrestrial ecosystem. Within the next fifty years, the human population is projected to double, and economic buying power for carbon-based products could triple. As there are no more unexplored frontiers, this increased demand from terrestrial ecosystem will have to be met with the existing natural resource base and in the face of a changing climate and extreme weather events. In addition, regional increases in soil erosion and atmospheric pollution could also have negative impacts on crop productivity and the natural resource base of the planet. With existing scientific knowledge it is impossible to accurately predict how these changes in the global climate may change the productivity of various crops worldwide and overall productivity of the terrestrial ecosystem. One way to deal with the complexity of the system, and its impact on crop productivity, is to develop and use mechanistic, process level computer simulation models, both at the field level and at the ecosystem level. This presentation outlines some examples of the development and use of the crop models for various applications to increase crop productivity and to mitigate the harmful effects of adverse environmental variables on natural resources, both in the current and in the future changing environment.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 97 Day 4: December 14, 2015
Concurrent Session L
Elevated CO2 ameliorates drought and high temperature stress effects in rice
Madan Pal*, Ashish K. Chaturvedi and Divya Shah Madan Pal Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Correspondence: [email protected]
Apart from rising CO2, high temperature and drought stress are important climate change factors, which are predicted to increase in frequency and magnitude under changing climate. These factors often interact with each other under field conditions and their resulting impacts may be different compared to individual exposure. This work is focused on physiological impact of combinations of these factors compared to independent exposure. The experiments were –1 conducted to analyse the role of elevated CO2 (eCO2, 550±34 μmol mol ) in combination with drought and high temperature stress on growth, gaseous exchange, carbon metabolism, grain yield and quality attributes in Nerica L- 44 and Pusa 1121 rice genotypes, raised inside FACE and open top chambers. Rate of photosynthesis, non-structural carbohydrates and ADPGase and starch synthase enzyme activity was stimulated under eCO2, decreased under drought and high temperature stress and reduction was less under combined exposure of eCO2 with drought or high temperature. Similar response was recorded for dry matter production and grain yield. Chalkiness in grains was more under eCO2 individually and further enhanced under combined exposure of drought and high temperature. Grain protein concentration decreased under eCO2 individually but reduction was less under combined exposure of drought and high temperature with eCO2. Amylose concentration in grains was less under combined exposure of drought and high temperature with eCO2 compared to exposure to eCO2 individually. These findings suggest that negative impacts of drought and high temperature stress on grain yield and quality in rice can be lowered if combined with eCO2 exposure.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 98 Day 4: December 14, 2015
Concurrent Session L
Rice adaptation pathways in response to heat stress at anthesis
Tanguy Lafarge*, C. Julia, N. Ahmadi and M. Dingkuhn
French Agricultural Research Centre for International Development, France Tanguy Lafarge *Correspondence: [email protected]
Coping with heat stress involves different options that account for the ability of plants to escape (early anthesis time), avoid (panicle cooling through transpiration) or tolerate (presence of genes of interest) heat at flowering. First, variability in the time of day of anthesis was correlated with the mean of climatic variables calculated for the 7-day period before flowering, over four distinct seasons and four contrasted varieties. The best predictive variables (negative correlations) were Tmin and VPD, with high values of both being associated with early times. Second, depending on conditions, panicle temperature varied between 9°C below and 2°C above air temperature at 2 m. A significant positive correlation was obtained between spikelet sterility rate and maximum panicle temperature at flowering, whereas no correlation was obtained with air temperature. By extrapolation, this correlation predicted minimal sterility with a panicle temperature of 3°C, and 50% sterility with a panicle temperature of 33-34°C. Third, a genome-wide association study of the sterility rate of 167 traditional and modern varieties (grown for six consecutive days at 37°C between 8 am and 2 pm at anthesis) detected 91 significant associations grouped into 12 independent regions located on eight chromosomes. The highest heat tolerance was detected for N22, an aus variety from India, and Peh Kuh, a traditional indica variety from Taiwan. Avoidance and tolerance appear as main genetic improvement pathways, whereas sensitivity of time of day of anthesis to air temperature and humidity seems already effective within the rice crop.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 99 Day 4: December 14, 2015
Concurrent Session L
Implications of climate change on plantation sector with special reference to coconut
K.B. Hebbar*, Ravi Bhat and P. Chowdappa K.B. Hebbar Plant Physiology, Biochemistry & Post Harvest Technology, ICAR-Central Plantation Crops Research Institute, Kasaragod-671124, Kerala, India *Correspondence: [email protected]
Plantation crops have a key role in the socio-economic development and transition from a subsistence agrarian economy to market oriented commercial cultivation due to their trade significance. Cultivation of plantation crops in India also has a rich diversity and varied history with each crop having its own distinct historical and economic context of development. The major plantation crops in the country include coconut, arecanut, oil palm, cashew, tea, coffee, rubber and several spice crops. Amongst these coconut is the predominant crop which provides sustenance to more than 10 million people and contributes Rs. 83000 million annually to the Gross Domestic Product (GDP) of the country. Most of these crops are grown either in hilly areas or coastal tracks where the threat of climate change is more. The impending climate change will affect coconut plantation through higher temperatures, elevated CO2 concentration, precipitation changes, increased weeds, pests, and disease pressure. As per the latest IPCC report rainfall becomes sporadic and temperature would rise. Coconut requires large volume of water (50 to 100 litre per day) for the production of highly productive canopy and with water limitation the canopy reduces and yield may reduce as high as 50% of irrigated gardens. In order to adopt the crop to climate change various soil, water and crop management techniques have been evolved at ICAR-CPCRI. The cropping and farming system models developed at the Institute involving grasses, vegetables, fruit trees, and animal component showed encouraging results in the farmer’s fields.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 100 Day 4: December 14, 2015
Concurrent Session L
Staygreen QTL3B is associated with improved grain and stover yield under postrainy sorghum growing conditions
H.S. Talwar1*, S.P. Deshpande2, Shiwesh Kumar1 and V. Vadez2 H.S. Talwar 1Indian Institute of Millets Research, Rajindranagar, Greater Hyderabad-500030, Telangana, India 2International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Patancheru, Greater Hyderabad-502324, Telangana, India *Correspondence: [email protected]
Postflowering drought is the major production constraint that adversely affects the crop growth and grain yield. Stay- green has been described as the best characterized trait contributing to the adaptation of sorghum to terminal drought conditions. The objective of this study was to assess the contributions of identified QTLs to grain and stover productivity under control and water-stressed conditions. Using B 35 as the donor for staygreen QTLs, introgression lines (ILs) were generated in two genetic background (S 35 and R 16). These lines along with local checks, recurrent parent and staygreen donor were evaluated for two years during the postrainy season and at four locations in India. A total of 42 introgressed lines (ILS) targeting stay-green QTL Stg1, Stg3A, Stg3B and Stg4 in R 16 genetic background, and 56 ILs targeting Stg1, Stg2, Stg3A, Stg3B, Stg4 and StgC in S 35 genetic background were tested under irrigated and terminal water-stressed (WS) treatments. Staygreen ILs had improvement in green leaf area retention at maturity (GLAM) in both R 16 and S 35 genetic backgrounds under both WW and WS conditions, and more so under WS condition. R16 was more responsive to the staygreen QTL introgression than S 35 in terms of GLAM, suggesting that effect of individual staygreen QTL introgression interacted with genetic backgrounds. Stg3B was the key staygreen QTL in enhancing the GLAM, particularly under WS conditions. Our results, established the superiority of stg3B in enhancing the Gy and Tdm accumulation over other individual QTLs or in combination under both the genetic backgrounds, particularly under WS conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 101 Day 4: December 14, 2015
Concurrent Session M
Control of meiotic gene expression in plants
I. Siddiqi*, S. Andreuzza, A. Singh, J. Davda, D.K. Singh, S. Pandey, Ramesha, A.V. Pardha Saradhi, A. Nalli, K.F. Max, S. Mahesh, S. Gharpure, V. Vijaybhaskar, V. Subbiah, V. Vinya, K. Bannerjee, S. Surendra, K. Bethoju and Y. Kavya Imran Siddiqi Centre for Cellular and Molecular Biology (CCMB), Hyderabad, Andra Pradesh, India *Correspondence: [email protected]
Meiosis produces haploid cells essential for sexual reproduction. Meiosis is a critical event in sexual reproduction. During meiosis, chromosomes recombine and segregate twice consecutively to produce haploid daughter cells, which differentiate into gametes. In humans, errors in meiosis are the leading causes of congenital birth defects. In plants, bypassing the meiotic program can lead to production of clonal seeds that retain hybrid traits that otherwise segregate. Thus, understanding the controls of meiosis has major implications for both health and crop improvement. In yeast, entry into meiosis activates transcription factors which trigger a transcriptional cascade that results in sequential co-expression of early, middle and late meiotic genes. However, these factors are not conserved, and the factors and regulatory mechanisms that ensure proper meiotic gene expression in multicellular eukaryotes are poorly understood. We have identified and characterized higher plant genes that control meiotic gene expression at different levels and through differential action in the two sexes. ACTIN RELATED PROTEIN6, a component of the SWR1 chromatin remodeling complex controls meiotic gene expression in the female lineage through loading of the histone variant H2A.Z. DUET/MMD1 controls expression of middle meiotic genes in the male lineage, and CDM1 an RNA binding protein controls levels of meiotic RNAs in the male lineage. Our results point to an integration of sex-specific gene expression programmes within a common meiotic programme leading to sexual dimorphism in plant meiosis and gametogenesis.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 102 Day 4: December 14, 2015
Concurrent Session M
Novel phenotyping technique for reproductive-stage salinity tolerance in rice
Mostafa Ahmadizadeh1,2, Naireen A. Vispo1, Cecilia Diana O. Calapit-Palao1,3, Iris Dawn 1 1 Pangaan and Rakesh K. Singh * Rakesh K. Singh
1Plant Breeding, Genetics, and Biotechnology Division, International Rice Research Institute, Philippines 2Department of Plant breeding, Sari Agriculture Science and Natural Resources University, Sari, Mazandaran, Iran 3Institute of Biological Sciences, University of Philippines, Los Baños, Laguna, Philippines *Correspondence: [email protected]
Rice responds differentially at different stages of development. The seedling and reproductive stages are the most sensitive growth stages with very weak association, suggesting that they are regulated by different processes and sets of genes/QTLs. There are hardly any studies exist on reproductive-stage salinity tolerance mainly because of the lack of reliable reproductive-stage-specific phenotyping techniques and incomplete knowledge of the stage-specific mechanisms of salinity tolerance. Two major challenges for screening exclusively for the reproductive stage are, 1) how to stress plants at the reproductive stage without stressing them at the seedling or late vegetative stage; and 2) how to impose the stress on different genotypes or mapping populations at equivalent growth stages of development because of the variability in the progression of growth rate through developmental stages. We standardise a methodology that allows salt translocation to the reproductive organs such as the flag leaf (the largest source for the sink) and panicle as quickly as possible just at the initiation of booting when the genotypes are at the same stage of tissue/organ development rather depending on age of the plant. Extra leaves were clipped, leaving only a minimum number of leaves needed by the rice plant that will not significantly affect grain yield and yield components. Salt stress equivalent to EC 10 dSm-1 was imposed on rice plants with trimmed leaves starting from boot leaf emergence up to 20 days in an experiment. Stage-specific effect of salt stress was verified by observing salt-sensitive and salt-tolerant genotypes. Details will be presented.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 103 Day 4: December 14, 2015
Concurrent Session M
Integrated genomics-assisted breeding strategy for rapid quantitative dissection of complex traits in crop plants
Swarup Kumar Parida1*, Alice Kujur1, Deepak Bajaj1, Shouvik Das1, Shailesh Tripathi2, 3 1 Hari D. Upadhyaya and Akhilesh K. Tyagi Swarup K. Parida
1National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India 2Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India 3International Crops Research Institute for the Semi-Arid Tropics, Patancheru-502324, Telangana, India *Correspondence: [email protected]
Development of genetically-tailored superior high-yielding stress tolerant (climate resilient) crop varieties is essential to improve the productivity and sustainability of global agriculture. However, most of these stress tolerance and yield- component traits targeted for crop improvement in current plant genomics and breeding research are complex and quantitative in nature, and multiplicatively governed by many genes/QTLs. The intervention of multiple modern structural, functional and comparative genomics approaches in classical/advanced breeding strategies i.e., “Genomics-assisted Breeding (GAB)” offers unprecedented opportunities to identify functionally relevant novel trait-associated major/ minor genes (QTLs) and elite rare alleles for precise molecular dissection of complex traits. This approach is also effective in studying the natural allelic interactions of favorable loci influencing trait variation in diverse crop genotypes that have adapted to varied agro-climatic conditions. More recently, the significance of GAB for dissecting complex genetic architecture of quantitative traits by implementing an integrated approach of association and QTL/eQTL mapping, associative transcriptomics and whole genome selective sweep-scans, has truly been realized. Essentially, the GAB expedites selection of desirable plants/recombinants with superior combinations of favorable alleles at multiple loci (gene/QTL pyramiding) specifically for trait genetic enhancement studies in crop plants via marker- assisted selection (MAS) and genomic/haplotype selection. The advent of various modern genomics and breeding tools, including sequence-based informative molecular markers, high-throughput marker genotyping assays and methods of rapid trait-specific gene/allele identification and their introgression into genotypes, have provided tremendous opportunities and flexibility to researchers for adopting diverse integrated approaches in genomics-assisted crop improvement program. This will greatly enrich our understanding of rapid quantitative dissection of complex traits for crop genetic enhancement through translational genomics (GAB and transgenics).
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 104 Day 4: December 14, 2015
Concurrent Session M
Molecular regulation of reproductive cold tolerance in chickpea genotypes
Harsh Nayyar1* and Kamal Dev Sharma2 Harsh Nayyar 1Department of Botany, Panjab University, Chandigarh-160014, India 2Department of Agricultural Biotechnology, Himachal Pradesh Krishi Vishva Vidyalaya, Palampur-176062, H.P., India *Correspondence: [email protected]
Chickpea is a cool-season food legume, which is sensitive to chilling temperatures (<20/10C), especially at its reproductive stage, leading to abortion of flowers to substantially reduce the podding potential. The underlying molecular mechanisms governing cold tolerance in reproductive components of chickpea are not known, which are imperative to be examined to address cold tolerance in this legume. In the present study, we investigated various physiological, biochemical and molecular mechanisms in leaves and anthers in chickpea genotypes contrasting for cold sensitivity. The findings revealed that maintenance of a stable sucrose metabolism in anthers is a vital mechanism affecting pollen development during cold stress in cold-tolerant genotypes. Investigations on the regulation of expression of differentially expressed genes in anthers of cold-tolerant genotypes under cold stress revealed two important features that govern cold tolerance in anthers i) differential expression of less number of genes and ii) up-regulation of majority of the differentially expressed genes. The main categories of genes governing cold tolerance in anthers were carbohydrate/triacylglycerol metabolism, signal transduction, pollen development and transport. Almost all the genes in these categories were up-regulated. Regulation of gene expression suggests that chickpea anthers use a dual cold tolerance mechanism wherein anthers sustain their development under cold by enhancing triacylglycerol and carbohydrate metabolism while pollen grains maintain normal development by regulating pollen development genes.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 105 Day 4: December 14, 2015
Concurrent Session M
Anthesis, anther dehiscence, pollen viability, stigma receptivity and pre, post-pollination biochemical changes in three varieties of mango
Kodthalu S. Shivashankara*, Tapas K. Roy and Gouribidanur A. Geetha K.S. Shivasankara ICAR- Indian Institute of Horticultural Research, Bengaluru, India *Correspondence: [email protected]
The study was carried out in order to evaluate the floral biology in terms of anthesis, anther dehiscence, pollen viability and in-vitro stigma receptivity as well as pre-, post-pollination biochemical changes in stigma of mango cvs. Alphonso, Totapuri and Amarapali, differing in fruit set intensity. Higher anthesis and anther dehiscence were observed before 8:00 AM and between 11:00 AM to 12:00, respectively in all the cultivars. As the temperature increased, anther dehiscence occurred early which wascoincided with higher anthesis resulting in higher fruit set. Higher stigma receptivity was observed within 3 hrs of anthesis in all the cultivars. The pH of the stigma was neutral during higher anthesis, anther dehiscence and also with higher stigma receptivity, and later on the pH was changed to acidic in all the cultivars. The stigmatic exudate was analysed to determine its components with a view to understanding their role in pollination and fertilization inmango.Free sugars (Sucrose, Galactose, Glucose and Fructose), free amino acids (Serine, Citrulline, Alanine, Proline, Phenylalanine and Glutamic acid) and hormones (Abscisic acid, Salicylic acid and Jasmonic acid) analysed using LCMS were consistently predominant and increased after anthesis. Among all the cultivars, Amarapali was observed to be good in terms of floral biological observations as well in biochemical parameters resulting in higher fruit set.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 106 Day 4: December 14, 2015
Concurrent Session M
Can one-sided sterility be regarded as a contrivance for sexual variability in Cleome viscosa?
Veenu Kaul* and Shveta Saroop Veenu Kaul Department of Botany, University of Jammu, Jammu-180006, J&K, India *Correspondence: [email protected]
Cleome viscosa L. belonging to family Cleomaceae is reportedly hermaphrodite. However, investigators have found the species to be quite complex in so far as its sexual system is concerned. Almost all floral features at structural and functional levels exhibit variation which quite likely leads to a flux in the breeding system of the species. The primary sex expression on an individual plant differs tremendously. Overall, six flower types differentiate on an individual plant. These can broadly be characterized as hermaphrodite, staminate (with rudimentary pistil), pistillate (with rudimentary stamens), male (with no pistil), hermaphrodite with staminodes and staminate with staminodes. The average production of these flower types varies from plant to plant and so does the frequency of the plants bearing them. Majority of plants are, however, and romonoecious with some exhibiting functional monoecy. This restriction is imposed due to the structural arrangements of sex organs while functional ones include stigma receptivity, pollen germination, ovular and pollen sterility. For-example female sterility is imposed by the non-functionality of ovules and male sterility due to pollen non-viability of shrivelled anthers. Interestingly, the one-sided sterility in context to the flower sex expression varies among plants subjected to simple manipulations of defoliation, deflowering and fruit removal. Results suggest major shift in resource investment towards the hermaphrodites further cluing towards the complex game of resource allocation influencing this sterility.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 107 ent Session M Young Scientist Session
YSS01 (IPPC0359) Plants actively avoid state transitions upon changes in light intensity: Role of light-harvesting complex II protein dephosphorylation in high light Nageswara Rao Mekala*, Mikko Tikkanen and Eva Mari Aro Department of Biochemistry, University of Turku Finland, Sirkkalankatu 8a B10, Turku-20520, Finland *Presenting author: [email protected]
Photosystem II (PSII) core and light-harvesting complex II (LHCII) proteins in plant chloroplasts undergo reversible phosphorylation upon changes in light intensity (being under control of redox-regulated STN7 and STN8 kinases and TAP38/PPH1 and PSII core phosphatases). Shift of plants from growth light to high light results in an increase of PSII core phosphorylation, whereas LHCII phosphorylation concomitantly decreases. Exactly the opposite takes place when plants are shifted to lower light intensity. Despite distinct changes occurring in thylakoid protein phosphorylation upon light intensity changes, the excitation balance between PSII and photosystem I remains unchanged. This differs drastically from the canonical-state transition model induced by artificial states 1 and 2 lights that concomitantly either dephosphorylate or phosphorylate, respectively, both the PSII core and LHCII phosphoproteins. Analysis of the kinase and phosphatase mutants revealed that TAP38/PPH1 phosphatase is crucial in preventing state transition upon increase in light intensity. Indeed, tap38/pph1 mutant revealed strong concomitant phosphorylation of both the PSII core and LHCII proteins upon transfer to high light, thus resembling the wild type under state 2 light. Coordinated function of thylakoid protein kinases and phosphatases is shown to secure balanced excitation energy for both photosystems by preventing state transitions upon changes in light intensity. Moreover, PROTON GRADIENT REGULATION5 (PGR5) is required for proper regulation of thylakoid protein kinases and phosphatases, and the pgr5 mutant mimics phenotypes of tap38/pph1. This shows that there is a close cooperation between the redox- and proton gradient-dependent regulatory mechanisms for proper function of the photosynthetic machinery.
YSS02 (IPPC0269) The role of inositol pyrophosphates in the perception of the plant defense hormone jasmonate Debabrata Laha* and Gabriel Schaaf Plant Physiology, ZMBP University of Tuebingen Germany, Auf der Morgenstelle 32, Tuebingen-72076, Germany *Presenting author: [email protected]
Diphosphorylated inositol polyphosphates, also referred to as inositol pyrophosphates, are important signaling molecules that regulate critical cellular activities in many eukaryotic organisms. In mammals and fungi, two distinct classes of inositol phosphate kinases mediate their biosynthesis: KCS1/IP6K- and VIP1/PPIP5K-like proteins. Recent studies in our lab show that PPIP5K homologs are widely distributed in plants and that Arabidopsis VIH1 and VIH2 are functional PPIP5K enzymes. We will report a specific induction of the inositol pyrophosphate InsP8 by jasmonate and will provide evidence that steady-state and jasmonate-induced pools of InsP8 in Arabidopsis seedlings depend on VIH2. We will further report a role of VIH2 in regulating jasmonate related defenses by potentiating jasmonate perception. Using in silico docking experiments and radioligand-binding based reconstitution assays we can show high affinity binding of inositol pyrophosphates to the F-box protein COI1–JAZ jasmonate co-receptor complex and will provide evidence that coincidence detection of jasmonate and InsP8 by COI1–JAZ is a critical component in jasmonate-regulated defenses.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 109 Young Scientist Session
YSS03 (IPPC0398) Transcriptomic insights into the plant structure and function: A parasitic plant and a unicellular alga Aashish Ranjan1*, Daniel Chitwood2, Brad Townsley3, Moran Farhi3, Yasunori Ichihashi4 and Neelima Sinha3 1National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India; 2Donald Danforth Plant Science Center, Saint Louis, USA; 3University of California, Davis, USA; 4RIKEN Center for Sustainable Resource Science, Japan *Presenting author: [email protected] Recent advances in the omics approaches, such as next generation sequencing in combination with genomics and transcriptomics, have facilitated the global insights into the genome and transcriptome to investigate the plant structure and function. Parasitic plants, which penetrate and establish vascular connections through specialized organs called haustoria to steal nutrients and water from host plants, are one of the most destructive agricultural pests. In order to understand the genetic basis of plant parasitism, we de novo assembled the transcriptome of an obligate stem plant parasite Cuscuta pentagona (Dodder) using RNAseq reads from various stages i.e. seed, seedling, vegetative stem, prehaustoria, haustoria and flowers. Subsequent gene expression analysis and dissection of transcriptional dynamics across the stages identified key genes and gene categories, such as plant defense and transporter genes, involved in the process of plant parasitism. The study is being used to identify potential gene targets for use in controlling infestation of crops by parasitic plants. In a parallel project, transcriptomics deciphered the molecular and genetic basis of patterning in one of the largest unicellular coenocytic alga, Caulerpa taxifolia, with distinct functional pseudo-organs. The study revealed a global, apical-basal pattern of the transcript distribution across the algal body. Interestingly, transcripts associated with specific cell compartments localize to organs with related functionalities. The results not only provided an intracellular atlas of transcript localization, but also demonstrated the contribution of transcript partitioning to morphology, independent from multicellularity, in plants.
YSS04 (IPPC0720) Physiological, metabolic, and transcriptional analysis of Arabidopsis to gain insight into plant responses to climate extremes Gaurav Zinta1,2*, Hamada AbdElgawad1, Ivan A. Janssens1, Gerrit T.S. Beemster1 and Han Asard1 1Department of Biology, University of Antwerp, Belgium; 2Shanghai Center for Plant Stress Biology, SIBS, CAS, China *Presenting author: [email protected]
Ongoing climate change involves co-occurring changes in various environmental factors, such as atmospheric CO2, temperature and precipitation. Extreme heat and drought limit plant growth and yield, whereas elevated CO2 alone is beneficial. Current climate change predictions indicate that climate extremes are not only projected to become more frequent and intense, but also to last longer. Molecular and physiological mechanisms underlying plant responses to climate extreme conditions under ambient and elevated CO2 are still poorly understood and little attention has been given to the temporal variation of plant stress responses, and on the transgenerational effects induced by climate extremes. We used Arabidopsis thaliana as a model system to understand plant responses to climate extremes. We investigated plant stress responses from whole-plant level, to the level of metabolites, enzymes, transcriptome and epi-genome. Main findings of this work are: 1) Elevated CO2 mitigates the impact of combined heat and drought, which could be explained by the higher up-regulation of antioxidant defense system (improved ROS scavenging) as well as by reduced photorespiration (less ROS formation) under high CO2, 2) Temporal metabolic profiling of plants exposed to combined heat and drought showed that short- and long-term stress exposure altered the plant primary metabolism differentially, and 3) Multigenerational exposure (for four generations) of Arabidopsis to heat stress resulted in more tolerant off springs. The descendants of the stressed plants showed a primed transcriptional state and greater up-regulation of defense related metabolites. Here, we expect the imperative role of epigenetic mechanisms in transgenerational stress memory of plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 110 Young Scientist Session
YSS05 (IPPC0091) Saltol QTL localized OsGATA transcription factor: An important candidate gene contributing salt tolerance in transgenic rice Kamlesh Kant Nutan1*, Sneh Lata Singla-Pareek2 and Ashwani Pareek1 1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected] Plant being sessile faces adverse environmental condition like drought, salinity, flooding and high and low temperature, which affects plant yield. Among them salinity is a major contributor of plant yield loss by abiotic stresses. One-third of the world’s population consumes rice as a staple food, but this crop is highly sensitive to salinity stress. In our study, we have studied differential expression of transcription factor genes localized within Saltol QTL of rice under salinity stress. Saltol QTL are present on short arm of chromosome number 1, accounts for more than 60% of the salt tolerance. On the basis of differential expression of 13 transcriptions factor genes (localized in Saltol) in contrasting cultivars of rice IR64 (sensitive) and pokkali (tolerant) under salinity stress, we have chosen GATA transcription factor. Constitutive overexpression of GATA transcription factor in transgenic rice leads to higher biomass accumulation as compared to wild type (WT) plants. Under salinity stress, transgenic rice seedling overexpressing GATA transcription factor shows higher salinity tolerance, high chlorophyll content and less ion leakage as compared to WT counterparts. Furthermore, at the cellular level, lesser damage of sub cellular organelles mainly chloroplast was observed in transgenic plants as compared to wild type and RNAi knock out plants. Osmolytes and other metabolites accumulation which help the plant in osmotic stress tolerance have been found to be higher in transgenic rice as compared to wild type. Our results suggest that GATA transcription factor may play a pivotal role in salinity stress tolerance in rice.
YSS06 (IPPC0171) Decoding the transcriptome and small RNAome to reveal genotype/developmental stage-specific molecular barcodes during drought and salinity stress in chickpea Rohini Garg* National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected] Drought and salinity are the major factors that limit chickpea production worldwide. In response to various stresses, plants evoke some common and stress-specific signature pathways that enable plants to discriminate between different stimuli and respond in unique ways. In the present study, we identified stress associated transcriptome of different chickpea genotypes under drought and salinity stress. RNA-seq of the root tissues of drought and salinity related genotypes were carried out under control and stress conditions at vegetative and/or reproductive development stages. Deep sequencing identified thousands of novel gene loci and alternatively spliced mRNA isoforms. A comparative analysis of the transcriptomes revealed divergent gene expression profiles in different chickpea genotypes at different developmental stages. A total of 79.1% and 84.8% transcripts exhibited genotype- and developmental stage-specific differential expression under drought stress and salinity stress, respectively. Overall, a significant fraction (~47%) of the transcription factor encoding genes showed differential expression under stress conditions. The key enzymes involved in metabolic pathways, such as carbohydrate metabolism, photosynthesis, lipid metabolism, generation of precursor metabolites and energy, post-translational protein modification, nucleic acid metabolism, cell redox homeostasis, cell wall component biogenesis and trehalose biosynthesis, were affected by drought and/or salinity stresses. Interestingly, different isoforms showed specificity of expression patterns across the chickpea genotypes and/or developmental stages as illustrated by the AP2- EREBP family members. We also identified genotype-specific and salinity regulated miRNAs in salt-sensitive and salt- tolerant genotypes of chickpea. These findings provide basis to dissect the gene regulatory network involved in drought and/or salinity stress response.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 111 Young Scientist Session
YSS07 (IPPC0651) Marker assisted recurrent selection (MARS) for drought tolerance using physiological and agronomic traits in bread wheat (Triticum aestivum L. em. Thell.) Harikrishna1*, N.K. Singh2, A.R. Rao3, Parveen Chhuneja4, P.C. Mishra5, S.C. Misra6, Niveditha Saini1, Kavitha Bhisth1, Arun Kumar1, Amitha C.R. Mithra2, Priyanka Vijay1, P.K. Singh1, Neelu Jain1, G.P. Singh1 and K.V. Prabhu7 1Division of Genetics, IARI, New Delhi-110012, India; 2National Research Centre on Plant Biotechnology, IARI, New Delhi-110012, India; 3Indian Agricultural Statistics Research Institute, New Delhi -110012, India; 4Punjab Agricultural University, Ludhiana, Punjab, India ; 5Jawaharlal Nehru Krishi Vishwa Vidyalaya, Powerkheda, India; 6Agharkar Research Institute, Pune, India; 7Joint Director Research IARI New Delhi-110012, India *Present author: [email protected]
The term drought carries networks of complex phenomenon. Because of its complex nature in terms of networks of pathways involved, complex inheritance and difficulties in phenotyping, breeding for drought tolerance is difficult. An effort was made to combine QTLs for drought adaptive traits using biparental population HI1500 X DBW 43; both parents carrying positive alleles for drought tolerance. Among 25 different traits phenotyped, NDVI at boot and grain filling stage canopy temperature at grain filling and milky stage, Leaf area index(LAI), grain weight per spike, thousand grain weight, biomass were showing significant correlation and regression values with yield under drought. These parents were diverse based on polymorphic 120 SSRs and 802 SNPs using 9K wheat SNP bead chip array. Based on validated QTLs information and multi-location (four location two year) testing of base populations 25 best individuals were selected for inter mating at F6 generation. Carefully chosen pairwise inter crosses (100) were attempted in order to combine QTLs using limited population size. A total of 17 loci were targeted and validation of molecular marker SSRs/QTLs present on 1B,1D,2D,2A,2B,2D,3A,3B,4A,5A,5B,6A, and 7B was done. Based on validated markers, wide range of inter crossed progenies in F5 generation with 4-12 positive alleles were obtained in first round of recombination out of 17 targeted loci segregating in the population. Genotypes which contains more number of QTLs out performed over checks in multi- location testing (four locations). These individuals will be subjected for further testing in co-ordinated trails at national level.
YSS08 (IPPC0195) OsGLYI-8 is a nucleus-localized glyoxalase I required for methylglyoxal metabolism in rice Charanpreet Kaur1*, Sneh Singla-Pareek2 and Sudhir Sopory2 1School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
The glyoxalase system generates D-lactate from methylglyoxal (MG), a cytotoxic byproduct of glycolysis which can otherwise increase to toxic levels under pathological conditions and form irreversible adducts with proteins and nucleic acids. The first enzyme of the glyoxalase system, glyoxalase I (GLYI), is usually a cytosolic metalloprotein that requires either Ni2+ or Zn2+ for its enzymatic activity. Plants have a variety of diverse GLYI isoforms that can play a crucial role during drought and high salinity. However, the exact subcellular localization, mechanism and physiological relevance of several of these diverse GLYI isoforms remain to be determined. Here, we analyzed a Zn2+-stimulated OsGLYI-8 from rice that has a so far unique nuclear localization and exhibits unusual biphasic steady-state kinetics. Further, loss of its homologue (AtGLYI2) in Arabidopsis leads to severe developmental defects on exposure to even low MG concentrations, much like yeast GLYI mutant which is also sensitive towards low MG concentrations. Our results thus, provide first evidence for MG metabolism in the nucleus and indicate towards a critical role of OsGLYI8 in plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 112 Young Scientist Session
YSS09 (IPPC0772) CBL-interacting protein kinase, CIPK9, differentially interacts and phosphorylates its upstream regulators, Ca2+ sensor CBLs in Arabidopsis Akhilesh K. Yadav*, Saroj K. Jha, Sibaji K. Sanyal and Girdhar K. Pandey Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi-110021, India *Presenting author: [email protected] Potassium is essential macronutrient required for various cellular processes and its deficiency leads to yield penalty in crops. Arabidopsis genome encodes 10 calcineur in B-like (CBLs) calcium sensors, which differentially interacts with 26CBL-interaction protein kinases (CIPKs). These CBL-CIPK complexes regulate various developmental, biotic, and abiotic as well as nutrient deficiency signaling processes. One of the CIPK family members, CIPK9 was identified as a critical regulator of K+ deficiency. In this study, we are discussing the detail biochemical regulation of CIPK9 activity, and phosphorylation of its interacting CBLs. CIPK9 specifically interact with CBL1, CBL2, CBL3 and CBL9 in yeast two-hybrid assay. The full length CIPK9 differentially phosphorylates its interacting CBLs in in-vitro kinase assay, hence specific CBLs act as physiological substrate of CIPK9.Autophosphorylation of CIPK9 is enormously increased specifically in presence of CBL2 or CBL3 while unaltered in the presence of CBL1 or CBL9. In contrast, CIPK9ÄR (CIPK9 without regulatory region i.e. having only kinase domain) also phosphorylates its interacting CBLs butitspreferential phosphorylation of CBLs is lost. Moreover, we found that mutated T178 to D178 residue in the activation loop of CIPK9 enhances kinase activity while T178 to A178 mutation reduces the level of autophosphorylation of CIPK9. However, in presence of CBL2 orCBL3, autophosphorylation of CIPK9, CIPK9T178D and CIPK9T178Aas well as transphosphorylation of CBL2 or CBL3 increases dramatically in in vitro kinase assay. Our study suggests that CBL1, 2, 3 and 9 interact differentially withCIPK9. Upon interaction CIPK9 can also differentially phosphorylate the CBLs and this differential phosphorylation of CBLs also influences the autophosphorylation as well as substrate phosphorylation activity of CIPK9 in a Ca2+ dependent manner in in-vitro kinase assay.
YSS10 (IPPC0327) Physiological and biochemical plasticity of Lepidium latifolium L. suggests it as Sleeper Weed in Western Himalayas under climate change scenario Tarandeep Kaur*, Hilal A. Bhat and Dhiraj Vyas Biodiversity and Applied Botany Division, Indian Institute of Integrative Medicine (CSIR), Canal Road, Jammu Tawi-180001, India *Presenting author: [email protected] Unlike the aggressive invasive character of Lepidium latifolium in riparian plains of North America, its growth is restricted in Ladakh Himalayas. In order to understand its future spread under climate change scenario in ecologically sensitive region of Western Himalayas, native populations of Lepidium latifolium in different microclimates were studied for physiological and biochemical plasticity. Different populations from sparsely populated site (SPS; 2807 m asl; 34.54 N, 76.13 E), moderately populated site (MPS; 4139 m asl; 33.20N, 78.64E) and densely populated site (DPS; 3164 m asl; 34.15N, 77.57E) were studied for their fluorescence, photosynthetic and biochemical characteristics. Results showed that the DPS had higher photosynthetic accumulation and transpiration rate despite lower VpdL and higher relative humidity. This suggests the regulation of its leaf temperature by evaporative cooling. Apart from its nutritive value, higher amounts of Sinigrin obtained in DPS might play an important role in water transport under environmental stress. The quantum efficiency of PSII photochemistry (Fv/Fm, NPQ, PSII) and light curve at various PPFD’s suggests better light harvesting potential and light compensation point at DPS than the other two sites. Concomitantly, plants at SPS had significantly higher lipid peroxidation, suggesting a stressful environment, and higher induction of antioxidative enzymes. High light intensities at MPS are managed by specialized contrive of carotenoid pigments and PsBs gene product. It is suggested that L. latifolium is present as ‘sleeper weed’ that has inherent biochemical plasticity involving multiple processes and its potential spread is linked to site-specific micro-environment, whereby, it would prefer flat valley bottoms with alluvial fills having high water availability under changed climate.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 113 Young Scientist Session
YSS11 (IPPC0162) Ambient ionization mass spectrometry imaging of rohitukine, a chromone anticancer alkaloid, during seed and seedling development in Dysoxylum binectariferum Hook.f (Meliaceae) P. Mohana Kumara1*, Amitava Srimany1, G. Ravikanth2, R. Uma Shaanker2 and T. Pradeep1 1DST Unit on Nanoscience and Thematic Unit of Excellence, Department of Chemistry, IITM, Chennai- 600036, India 2School of Ecology and Conservation, Deprt. of Crop Physiology, University of Agricultural Sciences, GKVK, Bengaluru-560065, India *Presenting author: [email protected] Chromone alkaloids are a unique group of compounds containing a flavonoid or a noreugenin chromone component linked with a ring containing one or more nitrogen atoms. These are sparsely distributed in plants. Rohitukine, a prominent chromone alkaloid, has been reported from only four species, belonging to the families, Meliaceae and Rubiaceae. Among them the Dysoxylum binectariferum (Meliaceae) is reported to accumulate the highest amount of rohitukine. In recent years, chromone alkaloid, such as rohitukine has gained wide attention because of their phrmacological activities. In fact, flavopiridol, a derivative of rohitukine, has been approved as an anticancer drug. The biosynthesis of rohitukine is not yet elucidated. In this study, we examine the spatial and temporal distribution of chromone alkaloids during various stages of seed and seedling development in D. binectariferum using desorption electrospray ionization mass spectrometry imaging (DESI MSI). Rohitukine (m/z 306.2) accumulation increased from early seed development to seed maturity stage. The spatial distribution of rohitukine was largely restricted to the cotyledonary tissue followed by the embryo and least in the seed coat. In seedling, root and leaves contained the high amount of rohitukine compared to the stem and twigs. Besides rohitukine, rohitukine acetate (m/z 348.2) and glycosylated rohitukine (m/z 468.2) were also detected, both through mass fragmentation and exact mass analysis. All of these molecules showed the tissue specific distribution of metabolites in seed, stem, root and leaves. In summary, DESI MS results indicated that spatial and temporal pattern of distribution of metabolites. It would be interesting to examine the underlying genetic basis for such a pattern.
YSS12 (IPPC0761) Biochemical characterization of an Arabidopsis thaliana cyclophilin AtCyp19-3 Gundeep Kaur1*, Supreet Singh1, Harpreet Singh1, Mrinalini Chawla2, Tanima Dutta1, Harsimran Kaur1, Kyle Bender3, W.A. Snedden3, Sanjay Kapoor2, Ashwani Pareek4 and Prabhjeet Singh1 1Guru Nanak Dev University, India 2Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India 3Queen’s University, Kingston, Canada 4Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected] Cyclophilins are ubiquitous proteins and constitute a multigene family in higher organisms. Several cyclophilins show peptidyl-prolyl cis-trans isomerase (PPIase) activity that catalyzes cis to trans isomerization, the rate limiting step in protein folding. Although Arabidopsis genome is predicted to contain 35 cyclophilin genes, biochemical characterization, imperative for understanding their cellular functions, has been reported only for few of these proteins. The present study reports that Arabidopsis cyclophilin, AtCyp19-3, is an enzymatically active protein and possesses PPIase activity that is specifically inhibited by CsA, with an inhibition constant of 18.75 nM. The catalytic efficiency of AtCyp19-3 for the substrate N-succinyl-ala-ala-pro-phe-p-nitroanilidine is 4.88 x 106 mol-1 s-1. Gel overlay and bimolecular fluorescence complementation assays provided evidence that AtCyp19-3 interacts with calmodulin in vitro and in vivo. The interaction of this protein with calmodulin is Ca2+- dependent. Further, the PPIase activity of AtCyp19-3 appears to be redox regulated and in silico modelling studies suggested that these mechanisms may be distinct in different members of this family, signifying specificity in their roles.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 114 Session A : Abiotic Stress Adaptation and Management
PA001 (IPPC0010) Mitigating effects of paclobutrazole on morpho-physiological, biochemical and antioxidant enzyme activities of mungbean [Vigna radiata (L.) Wilczek] under flooding stress A. Hemantaranjan* and D.K. Yadav Department of Plant Physiology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221005, India *Presenting author: [email protected]
An extensive investigation was conducted during kharif (rainy season) on the role of paclobutrazol (PBZ) on morpho- physiological, biochemical and antioxidant enzyme activity in flooding tolerant (HUM-16) and susceptible (HUM-12) mungbean (Vigna radiata L.) genotypes at pre-flowering stage. Paclobutrazol was applied both as seed soaking treatment (ST) and foliar spray (FS) (at pre-flowering stage). The flooding stress was carefully imposed at pre-flowering stage up to 7 days. Observations recorded revealed that paclobutrazol @ 10 ppm (ST) + 10 ppm (FS) significantly increased number of leaves per plant, fresh and dry weights of shoot and root, relative water content (RWC) and nitrate reductase activity (NRA) followed by antioxidant enzyme activity, viz., superoxide dismutase (SOD), ascorbate peroxidase (APX) and alcohol dehydrogenase (ADH) while, reduced plant height per plant and leaf area per plant in both the genotypes were also noted in PBZ treated plants under stress. Control plants without PBZ registered lesser significant values on the aforementioned parameters under flooding stress. Hence, findings elucidated that PBZ treatments positively influenced the aforementioned parameters. Nevertheless, the threshold concentration of PBZ @10 ppm ST + 10 ppm FS played significant role in mitigating the flooding stress by enhancing number of leaves per plant, fresh and dry weights of shoot and root; leaf relative water content, NRA, SOD, APX and ADH activities. Interestingly, the advantageous threshold level of PBZ as seed treatment combined with foliar spray treatment in this experiment was evaluated to be an economic source having considerably high potential as compared to other plant growth regulators and data for which shall be further discussed.
PA002 (IPPC0075) Abiotic stress tolerance in transgenic potato with reduced expression of PSII manganese stabilizing protein Chandrama Prakash Upadhyaya1* and Se Won Park2 1Department of Biotechnology, Dr. Hari Singh Gour Central University, Sagar-470003, Madhya Pradesh, India 2Department of Molecular Biotechnology, Konkuk University, Seoul, Korea *Presenting author: [email protected]
Manganese stabilizing protein (MSP) is an important component of the Photosystem II (PSII) oxygen evolving complex. We have reported the transgenic potato plants with reduced expression of MSP (named as MSP-As) and the response of MSP-As plants towards salinity, heavy metal and osmotic stresses. MSP-As plants treated with NaCl, ZnCl2 or mannitol solution showed significant level of tolerance under all the stress conditions. Specific enzyme activities of major ROS- scavenging enzymes were found significantly higher in MSP-As plants than the control plants. MSP-As plants accumulated increased levels of proline and low molecular weight metabolites such as ascorbate and α-tocopherol, which indicated that these plants were much more resistant to stress compared to the corresponding control plants. The primary photochemical efficiencies and the OJIP kinetics analyses further confirmed that MSP-As plants were in better optimal health under stress compared to the control plants. Although the exact reason behind the increased stress tolerance in stressed MSP-As plants is unclear, our results strongly indicate the role of MSP, which unknown function in abiotic stress tolerance.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 115 Session A : Abiotic Stress Adaptation and Management
PA003 (IPPC0085) Comparative performance of wheat genotypes adjusting by sowing windows and moisture regimes in Eastern Indo-Gangatic Plains S.K. Dwivedi1*, Santosh Kumar1, S.K. Singh1, J.K. Mishra1 and S. Singh2
1Division of Crop Research, ICAR Research Complex for Eastern Region, Patna, Bihar-800014, India 2International Rice Research Institute (IRRI), Manila, Philippines *Presenting author: [email protected] Understanding of physiological mechanisms that enable plants to adapt to water deficit stress and maintain growth and productivity during stress period could help in screening and selection of tolerant genotypes. Keeping these views in mind, an experiment was conducted at ICAR Research Complex for Eastern Region, Patna to study the comparative performance of five wheat genotypes (HD2987, K7903, HI1563, HD2824 and HD2967) grown under irrigated (at CRI) and completely rainfed conditions adjusted by sowing windows (15 Nov. & 30 Nov.). Study revealed that least impact of sowing dates was observed among genotypes in terms of physiological traits and yield attributes. The average yield reduction from irrigated to rainfed condition was 31 percent. Under rainfed condition, the higher yield was observed with genotype K7903 (1.9 t ha-1) followed by HD 2987 (1.8 t ha-1) as compared to other genotypes. Moreover, the physiological traits (relative water content; RWC, membrane stability index; MSI, total chlorophyll content and photosynthetic rate) were declined while proline content was improved under rainfed condition across the genotypes at both the developmental stages (anthesis and A+20 days). However, the genotype HD 2987 followed by K7903 were able to main high RWC (67.59 and 64.84%; 67.73 and 61.86%), and photosynthetic rate (15.70 and 10.73 μmol m-2 s-1; 17.76 and 10.74 μmol m-2 s-1) as compared to other genotypes at anthesis and post-anthesis stage, respectively. High RWC and photosynthetic rate is an important indicator to drought tolerance. This study will boost wheat production under completely rainfed condition.
PA004 (IPPC0090) Development of a robust screening protocol and, identification and characterization of novel heat tolerant rice mutants Priyanka Das1*, Ray Singh Rathor1, Sneh Singla-Pareek2 and Ashwani Pareek1
1School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Global warming is expected to have negative effect on plant growth due to the damaging effect of high temperature on plant development and yield. The increasing threat of high temperature might lead to catastrophic loss of productivity of vital food crops like rice. Therefore, it is necessary to develop heat tolerant rice plants by transgenic or mutation breeding approaches to overcome this problem. Mutation breeding is accepted as a powerful technique in the present era to obtain stress tolerant rice plants. We have developed a robust screening protocol and obtained three heat tolerant mutant plants from M6 mutant population of gamma-irradiated rice. High temperature tolerance is determined at first round of screening by phenotypic performance of mutant lines as compared to wild type after heat treatment and recovery. As indicators of heat tolerance at seedling stage, parameters such as leaf colour, root length, shoot length, fresh weight, leaf tip burning and ‘leaf tip kink’ were taken. In the second round of screening, physiological and biochemical analysis (lipid peroxidation, electrolyte leakage study and antioxidant enzyme assays) were carried out to authenticate the tolerance of mutants towards heat stress. Third round of screening of the heat tolerant mutant (confirmed at seedling stage) plants was carried out at flowering (during anthesis) stage following heat treatment and subsequent physiological and biochemical characterization. Our results showed that the putative heat tolerant rice lines are better in comparison to WT plants in terms of better phenotype, better enzymatic activities, better photosynthesis and higher yield.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 116 Session A : Abiotic Stress Adaptation and Management
PA005 (IPPC0100) Transgenic Arabidopsis thaliana over-expressing MuSMT1, a gene encoding sterol methyl transferase1 from Macrotyloma uniflorum improves tolerance to abiotic and biotic stresses Shikha Masand* and Sudesh Kumar Yadav Plant Metabolic Engineering Laboratory, Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061 (HP), India *Presenting author: [email protected]
Sterol methyl transferase1 (SMT1), a rate regulating enzyme catalyzes the production of 24-ethyl sterols. Transgenic Arabidopsis over-expressing MuSMT1 encoding SMT1 enzyme protein was isolated from Macrotyloma uniflorum and the beneficial effect of enhancing 24-ethyl sterol biosynthesis was studied. Transgenic Arabidopsis showed tolerance against multiple abiotic stresses such as high temperature, low temperature, drought, salinity, and oxidative stresses, as observed by retention of RWC, chlorophyll content and reduced MDA and H2O2 levels compared to WT plants. Transgenic Arabidopsis also showed significant resistance to Botrytis cinerea, a necrotrophic fungus due to enhanced accumulation of 24-ethyl sterols. Paclobutrazol, a potent inhibitor of sterol biosynthetic pathway, severely impaired plant growth by reducing 24- ethyl sterols production. The impaired growth of WT plants was rescued by the exogenous application of stigmasterol, suggested the requirement of 24-ethyl sterol for growth and development. Taken together, this study has for the first time documented the importance of 24-ethyl sterols accumulation in providing biotic and abiotic stress tolerance by MuSMT1 over-expression.
PA006 (IPPC0113) Elucidating molecular markers for the loss and re-establishment of desiccation tolerance in pea (Pisum sativum) seeds Balram Sahu* and Subhash Naithani Seed Biology Lab., SoS in Life Sciences, Pt. Ravishankar Shukla University, Raipur-492010, India *Presenting author: [email protected]
Most outstanding feature of dry seeds necessary for ex-situ storage is desiccation tolerance (DT). DT is the ability of an organism to withstand removal of its intracellular liquid water, then resume normal metabolism after rehydration and to survive in this state for ecologically significant period. The DT which is lost in the germinated seeds can be re-induced by the treatment of PEG, osmotic stress agent etc. Our study offers insight on the role of AOS, antioxidative enzymes and desiccation proteins [LEA and HSP] as a biomarker in pea seeds during loss and re-establishment of DT. The respiration capacity of germinated pea seeds declined with prolonged exposure of dehydration using silica gel. The loss of DT in the germinated pea seeds was negatively correlated with the accumulation AOS [superoxide and H2O2] measured in situ and in vivo. The accumulation of AOS was promoted due to impairment of antioxidative capacity [quantitative, isoenzyme and gene expression analysis of SOD, CAT and APX in the germinated pea seeds]. The pattern of desiccation tolerance protein [LEA and HSP’s] was similar to antioxidative enzyme. The re-establishment of desiccation tolerance in the germinated pea seeds was accompanied by reorientation of AOS, antioxdative system and DT-proteins thus confirming their vital role in the DT. The biomarkers of DT such as AOS, antioxidative enzymes and DT-proteins is discussed in loss and re-establishment of DT in the germinated pea seeds a model system developed for understanding the less known pathways of DT.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 117 Session A : Abiotic Stress Adaptation and Management
PA007 (IPPC0118) Moisture stress tolerance in lentil (Lens culinaris Medik.) genotypes in relation to physiological, biochemical parameters and 13C discrimination B.K. Mishra1, J.P. Srivastava1* and J.P. Lal2 1Department of Plant Physiology, 2Department of Genetics and Plant Breeding, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221005, India *Presenting author: [email protected]
Three microsperma (HUL-57, KLS-218, NDL-1) and five macrosperma (DPL-15, DPL-62, IPL-81, IPL-406 and L-4076) lentil genotypes were sown in field under irrigated and rainfed conditions. Drought sensitivity indices indicated microsperma types to be more tolerant to drought than macrosperma types. Pod formation stage was the most sensitive stage to drought. Stress reduced durations between different phenophases, total dry matter production and seed yield, but reduction was of lesser magnitude in drought tolerant genotypes. Efficient remobilization of assimilates and nitrogenous substances from vegetative parts to seeds, especially during later phase of crop growth, was observed in drought tolerant genotypes. In drought-stressed plants reduction in seed yield was mainly due to reduction in pods per plant and fruiting nodes per plant. Drought tolerant genotypes maintained higher relative water content (RWC), higher amounts of proline and soluble sugars, and lower amounts of malondialdehyde and H2O2 in leaves under stress. Drought tolerant genotypes exhibited lower 13C discrimination (Δ13C) and in normal plants Δ13C was associated negatively with seed yield, dry matter production and specific leaf weight (SLW) of plants under drought. Efficient remobilization of assimilates and nitrogenous substances from vegetative parts to seeds during later period of crop growth, maintenance of higher leaf relative water content and accumulation of osmolytes (proline and soluble sugars) were the parameters associated with moisture stress tolerance in lentil. It is suggested that Δ13C technique may be used to identify drought tolerant and susceptible lentil genotypes. Nevertheless, SLW under stress condition may be taken as a reliable alternative of Δ13C.
PA008 (IPPC0121) Trichoderma-colonization induced morpho-physiological and molecular amendment in drought stressed rice plant Veena Pandey1*, Mohammad Wahid Ansari2, Suresh Tula2, Ranjan Kumar Sahoo2, Gurdeep Bains1, Atul Kumar1, Alok Shukla1, Narendra Tuteja2 and Jitendra Kumar1 1Department of Plant Physiology, G.B. Pant University of Agriculture & Technology, Pantnagar-263145, Uttarakhand, India 2International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
The present study was carried out to assess the potential of Trichoderma harzianum to induce resistance under drought stress in two rice genotypes namely Pusa Basmati 1 and Pant Sugandh Dhan 17. Root-dip treatment of rice seedlings with Trichoderma formulation was given at the time of transplanting i.e. 21 days after sowing. The formulation was prepared by using Echinocloa frumentacae as substrate and the concentration of viable propagules was found to be 107 cfu/g. For determining the dose response, concentrated solution of Trichoderma, containing 10g/L (Dose1), 20g/L (Dose2) and 30g/L (Dose3) of Trichoderma formulation was prepared. Rice seedlings of all the genotypes were dipped in these three solutions for 25 min and thereafter planted in pots having sterilized soil. Seedlings dipped in distilled water were used as control. Drought stress was given at the time of flowering with withdrawn of water for five days. The soil moisture content in pots on fifth day was 8±2%. Results showed a significant amendment in the transcript levels of dehydrin and aquaporin genes in a dose responsive manner, as compared to control. Inoculation with Trichoderma induced prominent leaf rolling and reduced the incidence of leaf senescence and tip burn caused by drought stress in rice plant. Thus, providing evidence that T. harzianum colonization in rice plants can be considered as a promising strategy to mitigate drought stress. This strategy can be explored further so that it can be applied in field to eliminate undesirable consequences of drought stress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 118 Session A : Abiotic Stress Adaptation and Management
PA009 (IPPC0139) Raising stress tolerant rice for better yield and quality through genetic manipulation of cyclophilin Suchismita Roy1*, Sneh Singla-Pareek2 and Ashwani Pareek1 1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Cyclophilins are a set of ubiquitous proteins present in all subcellular compartments, involved in a wide variety of cellular processes. Cyclophilin 2 (OsCyp2) is a cytosolic member of the immunophilin family from Oryza and is a 519 bp gene encoding 172 amino acids. qRT-PCR data suggests that transcript abundance of OsCyp2 is regulated under different stress conditions. Localization studies confirm that OsCyp2 is localized in both cytosol and nucleus, indicating its possible interaction with several other proteins. Promoter analysis of this gene demarcates the presence of several stress related cis-acting elements, which did show inducibility of the promoter under different stress conditions. Stable overexpressing transgenic lines of this gene in Oryza sativa imparted tolerance towards multiple abiotic stresses as evidenced by higher germination efficiency, higher root length, more number of lateral roots, higher shoot length, chlorophyll content, and K+/ Na+ ratio as well as less calcium accumulation observed in the transgenic plants, under stress conditions. Transgenic plants also showed reduced MDA content and less electrolyte leakage under stress conditions than WT plants, suggesting better ion homeostasis in the transgenic plants than WT plants. RNAi lines of OsCyp2 gene validated these results with an observed phenotypic change in the seed morphology, seed weight and yield quality. Thus, it is proposed that overexpression of this gene is suitable for genetic engineering for better grain quality, yield and tolerance to cope with unfavourable conditions.
PA010 (IPPC0143) OsHK5, a histidine kinase of rice, working as multi-stress responsive gene in Saccharomyces cerevisiae Priyanka Gupta1*, Sneh L. Singla-Pareek2 and Ashwani Pareek1 1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, JNU, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Signal transduction cascades regulate cellular functions under normal and stress conditions. Two component signalling system (TCS) is one of the examples of signal cascades which play significant role in signal transduction in both prokaryotes and eukaryotes. TCS members include membrane bound histidine kinase (HK) as receptor and its cognate response regulator (RR) in cytosol. Plants have histidine phosphotransfer protein (HPT) as adaptor protein along with HK and RR. Perception of an environmental stimulus by membrane bound histidine kinase leads to autophosphorylation of conserved histidine residues. Signalling proceeds through transfer of the phosphoryl group to histidine residue of Histidine phosphotransfer protein (Hpt), which subsequently transfers phosphoryl group to aspartate residue of response regulators making a relay of phosphate transfer from His to Asp to His to Asp. In this study, we have investigated the role of rice histidine kinase, OsHK5 in abiotic stresses along with the rest of the sensory histidine kinases of rice. In silico analysis revealed that OsHK5 is a close homolog of Arabidopsis cytokinin receptor AHK2, and functions as membrane localised homodimer. OsHK5 is highly expressed throughout various developmental stages in rice. Expression of OsHK5 is induced in response to multiple abiotic stresses. OsHK5 along with the other OsHKs can functionally complement yeast Äsln1 temperature sensitive mutant. Yeast transformants carrying OsHK3b and OsHK5 can cope up with salinity, osmotic stress and high temperature. Assessment of growth of yeast transformants carrying OsHK5 in response to plant hormone cytokinin reveals that it may be functioning independent of cytokinin signalling.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 119 Session A : Abiotic Stress Adaptation and Management
PA011 (IPPC0167) Thermal imaging to assess genetic variation in drought adaptation of soybean cultivars Mahesh Kumar*, Ajay Singh, Susheel Raina, V. Govindasamy, R.L. Choudhary, Jagadish Rane and P.S. Minhas ICAR-National Institute of Abiotic Stress Management, Malegaon, Baramati-413115, Pune India *Presenting author: [email protected]
Soil moisture deficit is the most important factor limiting soybean yields in drought prone regions. With diminishing share of fresh water for agriculture this problem is likely to aggravate hence water use efficient varieties are essential to enhance and stabilize the production of crops like soybean. Though advances in genomics of this crop are remarkable, genes contributing to drought-tolerance are elusive mainly because of lack of precision in phenotypic characterisation of responses of large set of diverse genotypes. Hence non-invasive tools such as thermal imaging system to understand crop canopy temperature dynamics are recently gaining more attention. However, methods for employing these tools for acquiring the image, processing and analysis are not standardised. Hence to optimise methods for differentiating responses of soybean to drought, 32 diverse genotypes with similar phenology were selected from the pool of 174 collections screened in preliminary experiments. They were raised in field in RBD with 8 replications. More than 2500 thermal image were acquired at different growth stages of crop to assess the genetic variation in canopy temperature. Simple methods were developed to process the thermal image and analysis was carried out with customised software supplied with instrument. The method could identify superior and high yielding genotypes despite higher canopy temperature and also those which can keep their canopy cooler than local adapted cultivars. However, high productivity of NRC-7 despite relatively higher canopy temperature than other genotypes throughout growth period could be partially attributed to high net assimilation rate and quantum yield, indicated by chlorophyll fluorescence parameters.
PA012 (IPPC0169) Comparative effects of salicylic acid and its derivatives on the water relation, antioxidant defence mechanism, nitrate reductase, malate dehydrogenase, lipid peroxidation and yield of clusterbean (Cyamopsis tetragonoloba) under water stress N.S. Nathawat1,*, V.S. Rathore1, Birbal1, N.D. Yadav1 and R. Bhargav2 1ICAR-Central Arid Zone Research Institute, RRS, Bikaner-334004, Rajasthan, India 2ICAR-Central Institute of Arid Horticulture, Bikaner-334006, Rajasthan, India *Presenting author: [email protected]
Drought is an important factor influencing the growth and physiological characteristics of plants. Salicylic acid and its derivatives are known to affect various physiological and biochemical activities of plants. A field experiment conducted during rainy (kharif) season of 2014 at CAZRI, RRS, Bikaner, Rajasthan to evaluate the efficacy of salicylic acid (SA), thiosalicylic acid (TSA) and 5-sulfosalicylic acid (5-SSA) on growth, physio-biochemical parameters and yield of cluster bean under water stress condition. The foliar application of SA and its derivatives had 10-23% greater membrane stability index and 13-23% higher NAR than control. The application of SA and its derivatives had 11-40, 14-45, 13-30, 9-22% higher superoxide dismutase (SOD), ascorbate peroxidise (APX), nitrate reductase (NR), malate dehydrogenase (MDH) activity, whereas the level of lipid peroxidation in terms of malondialdehyde (MDA) content was 12-29% lower compared to water spray control. The MDA content in leaves varied from 8.9 to 12.7 μmol MDA g-1 fr. wt.; being lowest with 1.0 mM SA (29%) over WS. The activities of SOD, APX and NR were highest with 1.0 mM SSA over control. Exogenous application of SA and its derivatives brought 10-19 % improvement in seed yield, being highest with 1.0 mM SSA, followed by 1.0 mM SA and 1.0 mM TSA. Application of SA and its derivatives resulted in higher tissue water content; increased activity of antioxidant enzymes, decreased level of lipid peroxidation and membrane injury and it also protected nitrate reductase activity against inhibition under water deficit conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 120 Session A : Abiotic Stress Adaptation and Management
PA013 (IPPC0183) The effect of sowing dates on frost tolerance in faba bean (Vicia faba L.) Najeeb Alharbi1*, Kedar Adhikari2 and Helen Bramley2 1Plant Breeding Institute, Faculty of Agriculture and Environment, The University of Sydney, 7/48-52 St Hilliers Road AUBURN, Sydney 2144, Australia 2The University of Sydney, New South Wales 2006, Australia *Presenting author: [email protected]
Radiation frost is an important abiotic stress affecting grain yield in faba bean. The purpose of this study was to find frost tolerant genotypes and investigate whether frost damage can be managed by adjusting sowing time. A selection of 15 diverse faba bean genotypes were evaluated under three different sowing dates (16 April, 7 May and 26 May 2014) with two replications in a field condition at Narrabri in New South Wales, Australia. A visual 1-9 scale was used for scoring the frost damage at vegetative and reproductive stage by examining stem and pod, respectively. There was a highly significant difference among genotypes for frost tolerance at the vegetative stage, but sowing date and its interaction with genotype was not significant. There was no significant variation in pod damage among genotypes, however, the sowing time had a highly significant impact indicating frost can be managed by altering the sowing time. The highest damage was found in the early sowing date. Moreover, frost damage in stems and pods was not correlated with any of the morphological traits, such as leaf area, number of leaves and internodes length. In conclusion, frost tolerance can be improved at the vegetative stage by including more tolerant genotypes in the breeding program, but there was no variation for tolerance at the reproductive stage. Frost events are not predictable which might be a challenging task if sowing date alone is considered in frost management. Therefore, more tolerant genotypes need to be identified in the future.
PA014 (IPPC0189) Multigene engineering bridges the yield gap in rice under abiotic stress conditions Brijesh Gupta1*, Khirod Sahoo1, Ashwani Pareek2, Sudhir Sopory1 and Sneh Singla-Pareek1 1Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India 2School of Life Science, Jawaharlal Nehru University, New Delhi 110067, India *Presenting author: [email protected]
Increasing the yield of crops is an important objective for biotechnologists. Considering the multigenic nature of stress response in plants, gene pyramiding or multigene engineering, act as a promising approach to develop stress tolerant plants. Glyoxalase pathway enzymes, viz. glyoxalase I (GlyI) and glyoxalase II (GlyII), are the two enzymes required for glutathione-based cellular detoxification of a metabolically produced cytotoxin methylglyoxal (MG). A sodium proton antiporter contributes towards cellular ion homeostasis by sequestering excess Na+ ions. In the present study, we have shown that gene pyramiding using GlyI, GlyII and sodium proton antiporter significantly improves tolerance to multiple abiotic stresses in both tobacco and rice. The photosynthetic efficiency of triple (GlyI+GlyII+sodium-proton antiporter) transgenic tobacco and rice lines was significantly higher than double or single gene transgenic plants and they were able to flower and set more seeds under salinity and drought stress conditions. Further, as compared to wild type, single or double gene transgenic plants, the yield penalty was significantly lower in the plants expressing all the three genes under such stress conditions. These results suggest that strategies of pyramiding genes to remove the cytotoxic metabolites like methylglyoxal (by engineering glyoxalase system) and sequestration of sodium ions (by engineering ion transporters) may significantly improve stress tolerance limits. Future studies may utilize a similar strategy to generate stress tolerant varieties in other agronomically important crops as well.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 121 Session A : Abiotic Stress Adaptation and Management
PA015 (IPPC0193) OsCBSCBS3 is a stress-responsive ATP and/or AMP binding protein involved in abiotic stress adaptation in rice Ashish Subba1*, Thilini Ariyadasa1, Charanpreet Kaur2, Ashwani Pareek2 and Sneh Lata Singla-Pareek1 1Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India 2Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
CBS domain is a small protein module (approx. 60 residues long) present in various classes of proteins in all kingdoms of life. This domain has been identified to have binding sites for adenosine derivatives, thereby regulating the activity of associated proteins or domains. In plants, CBS domain containing proteins are differentially expressed during abiotic stress and few of them have been shown to be involved in stress adaptation, but still much is yet to be known about their physiological role in the system. Here, in order to understand the role of CBS domain containing proteins, we have cloned and characterized OsCBSCBS3 from rice which encodes a two CBS domain pair-containing protein and is differentially induced in response to abiotic stress conditions in salt-tolerant Pokkali and salt-sensitive IR64 rice cultivars. Our in silico studies revealed high similarity of OsCBSCBS3 to gamma regulatory subunit of AMP-activated protein kinase from different species. Further, subcellular localization studies indicated cytoplasmic localization of OsCBSCBS3. Notably, OsCBSCBS3 was found to possess binding affinity towards AMP and ATP, with higher affinity for AMP than ATP and could also hydrolyze ATP. Heterologous expression of OsCBSCBS3 in E. coli led to enhanced tolerance of bacterial cells towards salinity and oxidative stress. Likewise, over-expressing OsCBSCBS3 in plant system also imparted tolerance to transgenic tobacco and rice plants towards multiple abiotic stresses. We thus, propose that OsCBSCBS3 acts as sensor of cellular energy status by binding to AMP and/or ATP and is involved in stress adaptation in plants.
PA016 (IPPC0194) Physiology and proteome analysis of rice and its mutant under salinity stress Nita Lakra1*, Khalid Anwar1, Sneh L Singla-Pareek2 and Ashwani Pareek1 1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Salinity is one of the major environmental constraints that affect the growth and productivity of rice worldwide. Salt tolerant mutants are promising varieties for saline soil and they constitute the unique genetic stocks for gene discovery. We have generated several mutant lines by gamma-irradiation of IR64 and screened for their salt tolerance. In order to get better understanding of molecular pathways affected in rice mutant in response to salinity, we have studied their physiological characteristics and applied 2D-DIGE approach to explore the proteome changes associated with salt stress response. Physiological analysis showed that chlorophyll contents, relative water content and carotenoids were significantly higher in mutants under salinity. The level of anti-oxidative enzymes like POD, SOD, APX and CAT showed higher activity in mutant (D100-200) as compared to parent (IR64). Proteomics analysis showed that ~200 protein spots were reproducibly detected which were sequenced and identified using protein function databases. Gene ontology analysis showed that these proteins are involved in metabolism, protein synthesis, photosynthesis and stress response. Some of the proteins which showed specifically higher accumulation in mutant as compared to their parent (IR64) were identified as putative SOD, transcription factors, glycosyl hydrolase, nuclear transport factor, ribosomal proteins, fructose bis-phosphate and Rubisco small subunits etc. Few proteins were specifically up-regulated in IR64 and down regulated in leaves tissues of its mutant such as serine/threonine protein kinases, eukryotic translation initiation factor, OsFBX139 F box protein. The possible role of all the identified proteins in salt stress adaptation will be discussed in detail.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 122 Session A : Abiotic Stress Adaptation and Management
PA017 (IPPC0198) Impact of early and delayed transplantation of paddy on physico-chemical, pasting, cooking, textural and protein characteristics of milled rice Priyanka Paul1, Amardeep Singh Virdi1*, Parmeet Kaur, Gulshan Mahajan2 and Narpinder Singh1 1Department of Food Science and Technology, Guru Nanak Dev University, Amritsar- 143005, India 2Department of Plant Breeding & Genetics, Punjab Agricultural University, Ludhiana-141004, India *Presenting author: [email protected]
The present study aimed to evaluate the impact of early and delayed transplantation of paddy on physico-chemical, cooking, pasting and protein profiling of milled rice of four cultivars (PR114, PR116, PR118, and PAU201) harvested from early (5th June, 15th June) and delayed transplantation (25th June and 5th July) of paddy. At the time of active grain filling and seed maturation stages, early transplanted paddy experience an average minimum and maximum temperature of 32.8 and 22.3°C, against 30.8 and 15°C, respectively, by delayed transplanted paddy. Lightness (L*) value, protein content, and lipid content of milled rice was lower while, apparent amylose content, cooked grain hardness, final viscosity, and setback viscosity were higher in the delayed transplanted paddy. Amylose content was positively correlated with final viscosity, setback viscosity and cooked grain hardness and negatively correlated with breakdown viscosity and adhesiveness of cooked grains. Cooked rice of PAU 201 showed exceptionally higher adhesiveness and lower hardness as compared to other cultivars, which may be attributed to lower average amylose storage in PAU 201. Protein profiling showed that the accumulation of prolamin (18 kDa; 16 kDa and 14 kDa polypeptides) was reduced in the milled rice from PR114, PR116 and PAU 201 upon delaying the transplantation from 5th June to 5th July. The changes in milled rice, which were caused by delayed transplanting of paddy, were due to cool night air temperature that affected the accumulation of starch, protein and lipids, thereby, influencing the quality of milled rice.
PA018 (IPPC0203) Exploring the role of AtTAF4b in abiotic stress tolerance Paomipem Phazang1*, Ulrike Bechtold2, Philip M. Mullineaux2 and Neera Bhalla Sarin1 1School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2School of Biological Sciences, University of Essex, Colchester, UK *Presenting author: [email protected]
AtTAF4b is one of the TATA binding protein associated factors constituting the TAFIID, a general transcription factor. In Arabidopsis thaliana there are about 21 such TAFs. Recent findings show that each TAF regulates a distinct subset of genes in contrast to the earlier belief that they are required for the accurate transcription of all genes. While a large body of data is available on TFIID complex in mammalian systems, not much has been elucidated in plants. In this study we ventured to investigate the implication of overexpressing the AtTAF4b gene in plants for abiotic stress tolerance. The AtTAF4b was isolated from Arabidopsis thaliana and was overexpressed in Nicotiana tabacum under the control of CaMV 35S promoter. Stable integration of the transgene was confirmed by Southern Hybridization using the gene specific probe. The expression of the gene at the transcript level was also confirmed by RT-PCR. The transgenic lines of Nicotiana tabacum overexpressing the gene AtTAF4b showed a significant increase in tolerance to NaCl and mannitol stress as compared to the untransformed control plants. Although the exact mechanism of how the overexpression of AtTAF4b leads to multiple stress tolerance is not known, our preliminary findings suggest that AtTAF4b may play a role in regulating the abiotic stress tolerant genes. Further high through-put studies need to be carried out to have a clear insight into the function of the AtTAF4b gene in plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 123 Session A : Abiotic Stress Adaptation and Management
PA019 (IPPC0204) Evaluation of management strategies to mitigate adverse effects of waterlogging on productivity and quality of flue-cured tobacco (Nicotiana tabacum L.) M. Anuradha1*, D. Damodarreddy2, K. Sivaraju2 and T. Goplakrishnamurthy2 1Plant Physiology, ICAR-Central Tobacco Research Institute, Research Station, Kandukur-522101, India 2ICAR-Central Tobacco Research Institute, Rajahmundry-533105, Andhra Pradesh, India *Presenting author: [email protected]
In recent years tobacco crop has been facing a series of cyclonic rains leading to waterlogging and prolonged droughts coupled with high temperatures. It is predicted that the changes in the climate may lead to extreme weather conditions including floods and droughts or uneven and untimely rains in the future. These changes may result in reduction of productivity and quality of tobacco. Though tobacco, can withstand water deficit to a fair degree, it is known to exhibit vulnerability to excess-water or waterlogging. There is need to develop technologies to minimize the negative effects of waterlogging on productivity and quality of flue-cured tobacco. In this context, field and pot culture experiments were conducted to evaluate management strategies (Soil application of KNO3, Foliar application of KNO3, Hoagland solution, Salicylic acid, Putriscine, Kinetin) for their potential to alleviate waterlogging induced adverse effects on tobacco. The condition of waterlogging was imposed in both pot and field culture at 60 DAP for 48 hours and after relieving the plants from excess water the plants were treated with different management interventions. Leaves were harvested at maturity, cured and were analyzed for quality. In the pot culture experiment, soil application of KNO3, foliar spray of kinetin and putriscine performed better in terms of enabling plants to recover faster. As soil application of KNO3 found better, all the foliar spray treatments were again tested in combination with soil application of KNO3. Foliar spray of kinetin in combination with soil application of KNO3 could minimize the adverse effects of waterlogging. These treatments were tested in the field condition and the results revealed that soil application of KNO3 (5 g on either side of the plant by making holes) in combination with kinetin spray @ 50 ppm twice at 10 days interval could minimize the adverse impact of excess water stress on yield and quality to some extent in flue-cured tobacco.
PA020 (IPPC0206) Role of Glucose Auxin responsive gene GAX2 towards thermotolerance in model plant system Arabidopsis thaliana Mohan A. Sharma*, Brihaspati N. Shukla and Ashverya Laxmi Plant Signaling, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected] Glucose-Auxin and heat responsive gene GAX2 plays role in thermotolerance. The subcellular localization of GAX2 as determined using YFP was found to be in cytosol and nucleus. The expression levels of GAX2 was examined in glucose signaling mutants and it was found that expression of GAX2 was not dependent on HXK1 dependent and independent signaling pathway. Overexpression lines of TOR showed increased expression of GAX2 mRNA whereas overexpression and RNAi lines of KIN10 exhibited decreased and increased GAX2 expression. Data extracted from Genevestigator also showed reduced transcript levels of GAX2 in TOR RNAi lines which further confirmed the plausible involvement of energy mediated glucose signaling. Kinetics of GAX2 expression in response to starvation and energy enriched condition revealed the role of energy signaling in GAX2 regulation. Furthermore, Cis-regulatory elements analysis indicated that 942bp promoter of GAX2 contains several heat shock elements (HSE and CCAAT) and other stress-response elements. Quantitative measurement of chlorophyll content and seedling survival assay revealed stronger thermotolerance in GAX2 overexpression lines than wild type. Bio-computational analysis through Genevestigator showed reduced expression of GAX2 in HsfA1 quadruple mutant (KO-hsfa1a/b/c/d). Besides, overexpression of GAX2 exhibited up-regulation of those Hsp genes, which are also up-regulated by glucose and HsfA1a, which may lead to stronger thermotolerance. Taken together, these results suggest that glucose-mediated TOR kinase signaling control GAX2 expression level, which eventually regulates thermotolerance response in Arabidopsis seedlings.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 124 Session A : Abiotic Stress Adaptation and Management
PA021 (IPPC0210) Pseudomonas putida regulates physiological, biochemical and molecular mechanisms of Cicer arietinum L. plants for coping with short-term drought stress and recovery Charu Lata* and Shalini Tiwari Division of Plant-Microbe Interaction, CSIR-National Botanical Research Institute, Rana Pratap Marg, Lucknow-226001, India *Presenting author: [email protected]
Drought is one of the most important environmental stress factors that adversely affect plant growth and yield. However, some plant growth promoting rhizobacteria (PGPRs) are known to improve plant health and promote growth during abiotic stresses. Present study suggested the potential role of Pseudomonas putida MTCC5279 (NBRIRA) in drought stress amelioration in two contrasting chickpea varieties namely, cv. BG-362 (desi) and cv. BG-1003 (kabuli) under both in vitro and greenhouse conditions. Drought stress significantly affected various growth parameters, water status, membrane integrity, osmolyte accumulation, reactive oxygen species (ROS) scavenging ability and stress responsive gene expressions at all stress durations, which were positively attuned upon application of NBRIRA in both chickpea cultivars. When compared to control, NBRIRA inoculation also helped in better recovery of both chickpea types. Quantitative (qRT) PCR analysis showed differential expression of genes involved in transcription regulation (DREB1A and NAC1), stress response (LEA and DHN), ROS scavenging (CAT, APX, GST), and ethylene biosynthesis (ACO and ACS) in both desi and kabuli chickpea subjected to drought stress and recovery with or without NBRIRA. Our results thus indicate that NBRIRA confers drought tolerance in chickpea by altering various morphological, physiological and biochemical parameters, as well as by regulating differential expression of at least 9 stress responsive genes. To the best of our knowledge, this is the first report on detailed analysis of plant growth promotion and stress amelioration in one-month-old desi and kabuli chickpea subjected to short-term drought stress and subsequent recovery in the presence of a PGPR.
PA022 (IPPC0212) Synchronization of antioxidant defense machinery by facilitation of redox state equilibrium in Naringenin-treated and Funneliformis mosseae inoculated-Cicer arietinum L. genotypes under salt stress Priyanka Singla * and Neera Garg Department of Botany, Panjab University, Chandigarh - 160014, India *Presenting author: [email protected]
Under prolonged salinity, equilibrium between production and scavenging of reactive oxygen species (ROS) is overridden by oxidative burst in legume plants. Arbuscular mycorrhizal (AM) symbiosis as well as flavonoid application helps plants to thrive in saline soils by countering stress induced redox imbalance of plants. However, probable cross-talk between AM and flavonoid and their relative influence in upholding cellular redox stability in stressed plants is an aspect that has not been explored hitherto and thus necessitates its in-depth investigation. Consequently, fully factorial experiments were designed to investigate the potential role of naringenin (Nar, 4 ìM) and AM (Funneliformis mosseae) in transforming antioxidant network of two salt stressed (0, 40, 60, 80, 100 mM NaCl) Cicer arietinum L. genotypes (PBG 5, DCP 92-3). Despite increase in primary antioxidants, Na+ accumulation-induced ROS build-up was coupled with lowering of ascorbate/ dehydroascorbate (AsA/DHA), reduced/oxidised glutathione (GSH/GSSG) and flavonoid content, more significantly in DCP 92-3 than PBG 5. In Nar treated and /or AM plants, increased antioxidants efficiently attenuated oxidative loads, with maximum redox stability attained by +Nar+AM plants. Improved efficacy was related with elevated recycling of reduced glutathione and ascorbate alongwith improved flavonoid concentration, thereby facilitating higher H2O2 scavenging and subsequently higher O2•- dismutation. However, PBG 5 was more responsive to Nar treatment and/or AM inoculation and displayed better redox equilibrium than DCP 92-3. Study suggested that in-order to improve chickpea growth and curtail salt induced oxidative constraints; shifting redox towards more reduced form constitutes an important defensive mechanism adopted by Nar and F. mosseae.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 125 Session A : Abiotic Stress Adaptation and Management
PA023 (IPPC0213) Effect of high temperature stress on physiological attributes, antioxidative enzymes and expression of heat shock proteins in wheat (Triticum aestivum L.) Narendra Gupta1*, Aarif Khan2 and Radheshyam Sain1 1Rajasthan Agricultural Research Institute, Durgapura- Jaipur-302018, India 2Plant Biotechnology Centre, SK Rajasthan Agricultural University, Bikaner, India Presenting author: [email protected]
Experiment was conducted to study the effect of heat stress on various parameters in five diverse wheat genotypes at seedling stage. Seeds were sown in plastic pots in laboratory at 25±2°C. For heat treatment, 20 days old seedlings were exposed to 36°C/42°C for 6 hours. Results showed that the heat stress significantly reduced the dry weight and length of seedlings in all the genotypes. Chlorophyll content and membrane stability index decreased but proline content increased in all the genotypes under stress conditions. Among genotypes, Raj 4083 exhibited better heat tolerance, particularly at 42°C. Activities of SOD, CAT, APX and POX increased significantly in all the genotypes but per cent enhancement was maximum in Raj 4083 and minimum in PBW 502. In SDS-PAGE protein profiling, 15 polypeptides were recovered in control plants with slight variations in temperature treated plants of most genotypes. Proteins with molecular weights of 85 kDa and 15 kDa were constitutively expressed in all the genotypes but proteins of 45 kDa were expressed only in tolerant genotypes. Expression analysis of heat shock proteins, particularly hsp 90 further indicated the existence of tolerance mechanism in Raj 4083. Thus, using a multi parametric approach, Raj 4083 was found most tolerant and PBW 502 most susceptible to heat stress. The study also provides evidence that antioxidant enzymes like SOD, CAT, GPX, APX and heat shock proteins like HSP 90 have association with heat tolerance in wheat and thus can be used as efficient markers for heat tolerance at seedling stage.
PA024 (IPPC0214) Development of drought tolerant variety in potato: present status and way forward Devendra Kumar1*, J.S. Minhas2, V.K. Gupta1, S. Rawal1, Name Singh1 and Brajesh Singh3 1Crop Physiology Biochemistry & PHT, Central Potato Research Institute Campus, Modipuram-250110, Meerut, India 2Central Potato Research Station, Jalandhar, India 3Central Potato Research Institute, Shimla-171001, India *Presenting author: [email protected]
Potato is highly sensitive to water stress. Current potato varieties have been bred for optimum inputs viz fertilizers, irrigation, temperature, soil type etc. Minor scarcity of any input therefore, offers substantial challenge for sustaining the productivity. Considering future water scarcity, a programme to breed drought tolerant varieties was started in 2005. Besides field screening of existing varieties, hybrids and exotic clones; carbon isotopes discrimination, proline profile etc were also considered in the selection of parents. Established protocols were followed for breeding and selection of desired tubers traits in subsequent generations. Promising hybrids were evaluated for leaf area pattern, photosynthesis rate and associated parameters, antioxidant defense mechanism, leaf senescence, tuber yield and tuber defects under reduced water regimes (25 % less). Four seasons (2010-11 to 2013-14) data at Modipuram showed that hybrid WS/05- 146 had >33.0 t ha-1 mean tuber yield as compared to 27.9 t ha-1 in Kufri Bahar (popular high yielding variety) which is distinctly higher (>18%) under mild water stress conditions and recorded highest Drought Tolerance Index (DTI) and lowest Drought Susceptibility Index (DSI). Similarly under well watered conditions (control), it had 36.6 t ha-1 (>15%) mean tuber yield as compared to 31.6 t ha-1 in Kufri Bahar. Consequently, WS/05-146 has qualified for multi-location trials under All India Co-ordinated Research Project-Potato (ICAR) and will be evaluated during 2015-16 at Bhubaneswar, Deesa, Faizabad, Gwalior, Hisar, Kota, Pune and Raipur. Further, efforts are under way to enhance N economy through improved agro-technology.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 126 Session A : Abiotic Stress Adaptation and Management
PA025 (IPPC0215) In vitro drought induced changes in antioxidant systems of Rauwolfia serpentina Narendra Kumar*, Deepika Sharma, S.C. Shankhdhar and Deepti Shankhdhar Department of Plant Physiology, College of Basic Sciences and Humanities, GBPUA&T, Pantnagar, Uttarakhand, India *Presenting author: [email protected]
The present study conducted in department of plant physiology, GBPUA&T, Pantnagar to study the in vitro drought induced changes in antioxidant systems of (Sarpagandha) Rauwolfia serpentina. It is endangered woody perennial medicinal shrub, belongs to apocynaceae family, having various secondary metabolite as well as enzymatic and non enzymatic antioxidant to prevent various disease like high blood pressure, mental disease, sleeplessness and several other disease. Due to their poor seed germination percentage, tissue culture technique is very useful method for rapid multiplication and drought induced study. In our experiment the leaf derived callus induction and proliferation of R. serpentina (variety Cim- sheel) observed best in MS media supplemented with 2,4 D (2 mg/l) and BAP (0.5 mg/l). Drought stress was given to proliferated leaf derived callus under different concentration of PEG (1%, 2% and 3%) along with 2,4 D (2 mg/l) and BAP (0.5 mg/l). Proliferation of callus under different concentration of PEG observed in 15, 30 and 45 day after inoculation. Tissue water content was found to be decreased while enzymatic activity of antioxidants i.e. SOD and CAT was enhanced with increasing PEG treatments in comparison to control. In case of non enzymatic antioxidant like flavonoids content also increased with increasing level of drought in different interval of days. So above results indicate that Sarpagandha good source of antioxidant under drought stress conditions. The use of different concentration of PEG helps to measure the level of antioxidant and making the drought tolerant genotype through tissue culture technique.
PA026 (IPPC0221) Thermotolerance in wheat (Triticum aestivum L.) in relation to gibberellic acid Shivani Nagar1, R. Dhakar2, Neera Singh3, Ajay Arora1 and V.P. Singh1* 1Division of Plant Physiology, 2Division of Agricultural Physics, 3Division of Agricultural Chemicals, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Most of the wheat growing areas of the world experience water deficit and high temperature stress. Recent report of Intergovernmental Panel on Climate Change (IPCC 2014) has indicated an increase of 4.80C from pre-industrial temperature by the end of the century. It will result in exposure of reproductive and ripening phases of wheat to terminal heat stress, leading to severe drop in productivity. In the absence of adaptation and CO2 fertilization benefits, a one degree celsius rise in temperature alone could lead to a decrease of six million tonnes of wheat production. Paclobutrazol (PBZ) a plant growth regulator is inhibitor of gibberellic acid (GA) biosynthesis. PBZ also confers the thermotolerance to the crop plants. Hence, we hypothesize that lower level of GA should provide thermotolerance. Four wheat cultivars were sown at different dates to create temperature variation at reproductive phase. GA and PBZ were applied exogenously. Heat stress induced lipid peroxidation, membrane deterioration and reactive oxygen species production, and this trend was reversed by PBZ but not by GA. Activities of antioxidant enzymes like superoxide dismutase (SOD), glutathione reductase (GR), ascorbate peroxidase (APX) and catalase (CAT) were enhanced in all the cultivars. Application of PBZ enhanced activities of all these enzymes, whereas, GA enhanced the activity of SOD and APX only. We also studied the expression of heat shock factors (HSFs) and heat shock proteins (HSPs). Application of paclobutrazol enhanced the expression of HSF A1, HSP 101, HSP 70 and HSP 17.8. Among the cultivars highest expression of HSP’s and HSF’s was observed in
C-306. GA enhanced the endogenous level of GA, while PBZ decreased it. Endogenous GA3 levels in plants showed negative correlation with MSI, yield/plant, test weight and SOD activity and positive correlation with lipid peroxidation, ascorbate peroxidase and H2O2 content.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 127 Session A : Abiotic Stress Adaptation and Management
PA027 (IPPC0224) In vitro drought induces oxidative stress and its impact on phenol and flavonoid content in callus of Withania somnifera Deepika Sharma, Narendra Kumar, Shailesh Chandra Shankhdhar and Deepti Shankhdhar* Plant Physiology College of Basic Sciences & Humanities, G.B. Pant University of Agriculture & Technology, Pantnagar-263145, Uttarakhand, India *Presenting author: [email protected]
An important herbal plant Withania somnifera (genotype J 20) was evaluated for its growth, water content, necrosis, phenol and flavonoid changes in callus tissue under PEG induced drought. Leaf derived callus were initiated on Murashige and Skoog medium supplemented with 1.5 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D), 0.20 mg L-1 kinetin (Kn), and subcultured on same media containing different concentrations of Poly Ethylene Glycol (1-3%). After 4 weeks of culture, callus growth and water content were determined. The results showed that increasing PEG concentration in the medium causes a gradual decrease in growth, water content of callus and create necrosis. However, in between the black necrotic callus, whitish embryogenic mass of cells were observed. The tolerant cells were again proliferated on stress media and used for further study of phenol, flavonoid and MDA content. It was observed that higher concentrations of PEG along with longer exposure (up to 45 days) induces oxidative stress in surviving callus and resulted in increased phenolics, flavonoid and malondialdehyde (MDA), which was positively correlated with an increase in the accumulation of PEG. Increasing concentration of phenols and flavonoids in the tissue act as scavengers of free radicals and also prevent their formation by chelating metals.
PA028 (IPPC0230) Isolation and characterization of SOS pathway genes from a halophyte, Oryza coarctata Showkat Ahmad Ganie* and Tapan Kumar Mondal Division of Genomic Resources, National Bureau of Plant Genetic Resources, IARI Campus, Pusa, New Delhi-110012, India *Presenting author: [email protected]
Salinity is an important abiotic stress which limits the agricultural production world-wide. Rice is regarded as a traditionally salinity sensitive crop. However, one of its wild relative species, Oryza coarctata, is highly salt tolerant (halophyte) which grows in the coastal regions under high saline conditions. It has been demonstrated previously that genes of O. coarctata can induced salinity tolerance in other crop plants including rice. Thus, we cloned full length sequences of SOS pathway genes encoding SOS1, SOS2 and SOS3 from this halophyte. The ORFs were found to be 3353bp, 1370bp and 633bp for SOS1, SOS2 and SOS3 respectively. The deduced amino acid sequences registered cAMP/cGMP binding domain, protein kinase domain and Ca2+ binding domain respectively which showed high degree of homology with corresponding genes of angiosperm. We further studied three dimensional model of all the genes and quality of predicted models were analysed with PROCHECK. Molecular docking of model revealed similar binding pockets for different substances and products. All the transcripts were found to salinity stress responsive indicated by qPCR study. Finally, a comparative analysis of these gene sequences from susceptible and tolerant glycophtes as well as facultative and obligatory halophytes of individual genotypes of Oryza species revealed interesting differences that might be responsible for differential salinity adaptation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 128 Session A : Abiotic Stress Adaptation and Management
PA029 (IPPC0246) Simultaneous over-expression of Eif4A and Lea2 genes in rice (Oryza sativa cv. PB1): Linking salt and drought tolerance Aakrati Agarwal*, Yashwanti Mudgil, K. Chandrasekhar, Subramanyam Chinreddy, Dhirendra Fartyal, Donald James and M.K. Reddy Plant Molecular Biology, International Centre for Genetic Engineering & Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Abiotic stress is a huge roadblock in crop-plant productivity worldwide especially in the case of rice (Oryza sativa). Drought and salinity are the most prominent abiotic stresses and are responsible for the major yield loss in rain-fed rice cultivation. Both these stresses are thought to be linked together as they lead ultimately to osmotic stress and the plant responses in both cases are similar. We developed a strategy to combat against these stresses simultaneously by using Lea2 and Eif4A genes. Lea2 gene is a late embryogenesis abundant protein, which specifically protects plant integrity against water loss. Eif4A has a reported role in providing salinity tolerance. To validate their relationship in rice, both genes were over-expressed simultaneously in indica rice plant variety PB1. The construct harbouring these two genes in addition to a plant selectable marker gene hptII and Bar was transformed into rice calli with a virulent Agrobacterium strain EHA105. The calli were selected in hygromycin B and the presence and stable integration of transgene in putative transgenic plants was confirmed by PCR for Lea2, Eif4A, Bar and hptII genes and Southern hybridization, which revealed the copy number in different transgenic events. Transgenic lines having a single and double copy number were selected for physiological analysis showing resistance to phosphinothricin. Simulated drought and salinity stress assays revealed an interdependent relationship between the two stresses.
PA030 (IPPC0253) Cross-genus role of endophytic microorganisms on growth promotion and metal phytoremediation in an arsenic accumulator terrestrial plant Solanum nigrum Gairik Mukherjee* and Anindita Seal Department of Biotechnology, University of Calcutta, Kolkata-700019, West Bengal, India *Presenting author: [email protected]
Endophytes are microorganisms that reside normally inside plant tissues and have been shown to have beneficial impact on the plant. Many of them are known for their growth promoting effects on the host as well as ensure their survival under various stress conditions. Among several plants growing in an arsenic contaminated site in West Bengal, a shrub Solanum nigrum was found to accumulate arsenic in its shoot with a transfer factor greater than 1. When grown under laboratory conditions, the plant was found to accumulate significant levels of arsenic with a transfer factor greater than 3. Therefore, the plant was chosen to be a suitable terrestrial model for arsenic phytoremediation. Lantana camara is an invasive weed that can survive environmental extremes. As such, seven arsenic tolerant endophytes were isolated from L. camara growing in the arsenic contaminated site. All endophytes had phosphate solubilization potential and produced the growth hormone auxin. One of the endophytes could even grow in nitrogen-free medium. All these properties are indicative of their probable growth promoting effects. When applied to Solanum nigrum under arsenic treated conditions, the endophyte consortium of L. camara was found to exhibit growth promoting effects on the plant. The consortium was also found to increase the leaf chlorophyll content related to growth promotion by enhanced photosynthesis. Besides, application of the consortium resulted in enhanced removal of arsenic from the soil and increased root to shoot arsenic translocation signifying suitability of the system in microbe- assisted phytoremediation of arsenic.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 129 Session A : Abiotic Stress Adaptation and Management
PA031 (IPPC0258)
Impact of high temperature and elevated CO2 in blackgram genotypes (Vigna mungo (L.) Hepper) H. Vijayaraghavan*, M. Pandiyan and C. Partheeban Department of Crop Physiology Tamil Nadu Agricultural University Coimbatore-641003, India *Presenting author: [email protected]
Investigation was carried out to screen the black gram genotypes for high temperature tolerance and to evaluate elevated
CO2-high temperature interaction. A protocol for rapid screening for high temperature was also developed namely, the temperature induction response (TIR) technique. Evaluation of black gram genotypes for heat tolerance under natural field condition indicated that six genotypes, VBG-06-002, VBG-06-005, VBG-06-010, VBG-07-001, VBG-10-008 and COBG-759 have highest yield components than the rest of the genotypes. These six genotypes fall under the category of heat tolerant, eight genotypes under moderately heat tolerant and five under the heat susceptible group. TIR at seedling stage exactly matched with that of field experiment as well as studies conducted under controlled environment chamber, suggesting the usefulness of TIR for screening for heat tolerance. The impact of elevated carbon dioxide and temperature on select six black gram genotypes was evaluated in the carbon dioxide chamber. The results showed that carbon dioxide concentration at 700 ppm under ambient temperature seems to be optimum in producing more number of flowers with consequent high fertility coefficient. Interestingly the two genotypes, VBG-06-002 and VBG-07-001 were identified as highly heat tolerant ones and deserve further research. These genotypes are being forwarded to the breeding programs and for further progeny selection.
PA032 (IPPC0275) Sorghum improvement for abiotic stress adaptation and climate change resilience in dryland conditions S.S. Rao1*, H.S. Talwar1, B. Subbarayaudu1, N. Kannababu1, C. Aruna1, K. Hariprasanna1, T.G.N. Rao1, Prabhakar Bhat2, P.V.V. Prasad3 and J.S. Mishra4 1ICAR-Indian Institute of Millets Research, Rajendranagar, Hyderabad-500030, India 2Centre on Rabi Sorghum (IIMR), Solapur, Maharashtra, India 3Kansas State University, KS, Manhattan, USA 4ICAR-ICAR Research Complex for Eastern Region, Patna, Bihar, India *Presenting author: [email protected]
Sorghum (Sorghum bicolor (L.) productivity is reduced due to the occurrence of drought and heat stresses in pre-and post flowering stages. Current research aimed at 1) quantifying the impact of drought (D), high temperature (HT) and combined stress (D*HT) on sorghum physiological traits and 2) assessing the climate effects on bioenergy sorghum quality. Treatments imposed in growth chambers 1) control [Irrigation + Optimum temperature (OT)(32/22 degrees C day and night, resp.]; 2) drought stress [drought for 20 days + OT (32/22 d C)]; 3) HT stress [Irrigation+ HT (38/28 d C)]; and 4) combined stress [drought + HT (38/28 d C)]. Leaf area decreased by 25% in D *HT stress over OT. Average leaf temperature increased by 8.0, 5.0, 2.5% resp., in combined, HT and D over OT. Root length and root -shoot ratio significantly increased by 11.0% in combined stress over optimum. Leaf water content decreased by 16.1% and 35.0% respectively, in D and D*HT over OT. Across rainy season plantings, sweet sorghum CSH22SS produced 29.0% more bioethanol than SSV84, while SSV74 yielded 16.6% more than SSV84. CSH22SS produced higher stalk, grain, sugar and bioethanol yields over SSV84 or SSV74. Both CSH22SS and SSV74 were more stable across plantings and showed climate resilience. In post rainy season, stalk yield decreased by 26-36% in late plantings over early. Based on stalk yield and sugar quality, PAC52093 and CSV19SS were stable across plantings and were climate adapted. The implication of these results is discussed for genetic enhancement of sorghum for climate change resilience.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 130 Session A : Abiotic Stress Adaptation and Management
PA033 (IPPC0276) Dehydration-responsive extracellular matrix of chickpea identifies CaFer1, a secretory ferritin that promotes stress tolerance and growth Shaista Parveen*, Deepti Bhushan and Niranjan Chakraborty National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author:[email protected]
Plants strictly regulate iron homeostasis to avoid iron deficiency which can impair key metabolic functions, and iron excess that produce damaging reactive oxygen. As key components of iron homeostasis, ferritins can store up to 4500 Fe (III) atoms in its central cavity and can release the stored iron when the need arises. However, the functions of phytoferritins, particularly in stress responses, remain poorly understood. In a previous proteomics study, we identified an array of stress-responsive proteins in the chickpea extracellular matrix that include ferritin. Here we describe the structure, expression and stress-responsive function of the chickpea ferritin, designated CaFer1. Water-deficit conditions trigger CaFer1 expression indicating its dehydration-responsive function, besides its role against Fusarium oxysporum infection. Functional complementation of the yeast frataxin-deficient mutant Deltayfh1 indicates that CaFer1 also functions in oxidative stress. CaFer1 expression increases in response to iron deficiency, and CaFer1-overexpressed seedlings showed altered expression of iron-responsive genes and improved growth and development. The mechanism of functions of secreted ferritins is well characterized in insects and animals, but no secreted ferritin has yet been identified in plants. The compartmentalization of CaFer1 to the extracellular matrix and the secretome, reported here for the first time, indicates that its function differs from that of other phytoferritins. CaFer1 interacts with the iron transporter IRT1, suggesting a distinct role in maintenance of iron homeostasis. Our results suggest that CaFer1 plays key role in iron-buffering and stress tolerance.
PA034 (IPPC0277) Influence of stress mitigating compounds on biophysical parameters and yield attributes under drought in groundnut (Arachis hypogeal L.) R.P. Patil1*, M. Harish1, A. Amaregouda1, M.M. Dhanoji1 and M.R. Umesh2 1Department of Crop Physiology, 2Department of Agronomy, University of Agricultural Sciences, Raichur-584133, India *Presenting author: [email protected]
Field experiment was conducted during the year 2014-15 at college of Agriculture, Raichur to investigate the appropriate technology to overcome the ill effects of water stress on growth and development of groundnut crop. Water stress plots were treated with various stress mitigating compounds, significantly higher pod yield was recorded with application of tricontanol @ 2.0 ml/l (28.3 q/ha) followed by KCl @ 2.5% (26.9 q/ha) methanol (2%). Biophysical parameters TR were shown maximum in tricontanol and minimum diffusive resistance was seen in same treatment. Whereas, among the stressed plot RWC was found more in tricontanol followed by KCl (76.6%). At 45 DAS, Finally yield deciding parameter in photosynthetic rate was showed significant differences between the treatments. Photosynthetic rate (31.87 μmol CO2 m-2 s-1) was higher in unstressed plot wherein continuous irrigation without stress. Among the different stress mitigating -2 -1 compounds foliar application of tricontanol has recorded significantly higher photosynthetic rate (30.13 μmol CO2 m s ) -2 -1 -2 -1 followed by Nitrobenzene (30.03 μmol CO2 m s ) Methanol (28.33 μmol CO2 m s ) Salicylic acid, Kaolin and lower in stressed plots. Finally the yield components No. of pods plant-1, Pod weight (g), pod yield (q ha-1) and harvest Index (%) were studied and maximum yield was recorded in the foliar spray of tricontanol (28.3 q ha-1) followed by KCl (26.9 q ha-1) and methanol (6%) (26.6 q ha-1).
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 131 Session A : Abiotic Stress Adaptation and Management
PA035 (IPPC0281) Screening of cotton (G. hirsutum) genotypes for water stress tolerance underrainfed conditions D.G. Dalvi*, D.B. Deosarkar and K.S. Baig Department of Agricultural Botany, Vasantrao Naik Marathwada Agricultural University, College of Agriculture, Parbhani-431402, Maharashtra, India *Presenting author: [email protected]
A field experiment was undertaken during kharif season for the period of eight years (2007-08 to 2014-15) with an object to screen cotton genotypes for water stress tolerance under protected and unprotected conditions, 155 genotypes of cotton (G. hirsutum) were screened, out of which the genotypes viz., KH-138, Sahana, NH-615, PH-1009, RCR-102, HBB-101, GTHV-02145,CPD-817, NH-630, WGHH-411, GTHV-0135, BS-30, ARBH-813, NH-635, GBHV-175, PH-1075, CSH-1111, GBHV-162 WDLH-1938, RHC-0717, TSH-04-115, RAN-806, BS-37 and BS-39 were found tolerant to water stress under rainfed conditions on the basis of physiological parameter i.e. high degree of relative water content (%), proline content (%), leaf area index, specific leaf weight and stress indices viz., dry matter stability index, leaf area stability index, plant height stability index, yield stability index with least (<1.0) drought susceptibility index (S) and chlorophyll stability index. Similarly, these genotypes also recorded highest seed cotton yield (kg/ha) with least reduction (%) in yield under rainfed condition. These genotypes may be used for breeding purpose in further development of drought tolerance genotypes in cotton.
PA036 (IPPC0290) Association of ascorbic acid with drought tolerance in wheat Suman Roy1*, Ajay Arora2 and V.P. Singh2 1ICAR-Vivekanand Parvatiya Krishi Anusandhan Sansthan, Almora-263601, Uttarakhand, India 2Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Wheat is one of the premier cereal crops in India but confronts a severe threat to sustainable production due to water deficit stress. Plants have evolved various strategies to overcome the adverse effects of such stresses. One of the strategies developed by plants to fight such stress includes generation of antioxidants as a response to stress. Evidences confirm that ascorbic acid (AA), a non enzymatic antioxidant, is one of the major antioxidant that respond to both biotic and abiotic stresses. The biological activity of AA is defined by its oxidation and subsequent regeneration into the reduced form. However, phenotyping of genotypes on the basis of any antioxidant has not been done till date. In this study, we have screened 30 wheat genotypes on the basis of endogenous reduced ascorbic acid content under water deficit conditions. A wide genotypic variability for endogenous reduced ascorbic acid content was observed under control and stress condition. Endogenous reduced ascorbic acid content decreased under stress conditions in most of the genotypes. Results of Drought Susceptibility Index (DSI), Relative Water Content (RWC) and Membrane Stability Index (MSI) confirmed that susceptible genotypes have a lower content of endogenous reduced ascorbic acid as compared to the tolerant genotypes. In addition lower endogenous reduced ascorbic acid under stress condition was due to restriction of stress induced hydrogen peroxide levels. Endogenous levels of the other antioxidants may provide a clear picture which can be used as a trait for phenotyping genotypes under water deficit condition.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 132 Session A : Abiotic Stress Adaptation and Management
PA037 (IPPC0293) Overexpression of a stress responsive MuWRKY3 transcription factor leads to enhanced drought tolerance in groundnut (Arachis hypogaea L.) K. Kiranmai, A. Nareshkumar, V. Amarnatha Reddy, O. Sudhakar Babu, G. Lokanadha Rao, U. Lokesh, B. Venkatesh and Chinta Sudhakar* Plant Molecular Biology Unit, Department of Botany, Sri Krishnadevaraya University, Anantapur-515003, India *Presenting author: [email protected]
WRKY transcription factors are plant specific transcription factors regulates several developmental events and stress responses in plants. The WRKY domain is defined by the conserved amino acid sequence named WRKYGQK at its N- terminal end and Zinc finger like motif. WRKY genes often respond to several stress factors which may act as negative or positive regulators. The aim of present study is to clone, characterize and overexpress WRKY3 gene from horsegram (Macrotyloma uniflorum) a dry-land drought tolerant pulse crop. To this direction, we have successfully amplified MuWRKY gene from horsegram using cDNA with heterologous primer sequences (#EU375350.1). PCR product was purified and cloned into pTZ57R/T vector and sequence confirmed. The nucleotide sequence of the full-length CDS (MuWRKY3) comprised 1476 bp; the deduced protein showed 491 amino acids with an apparent molecular weight of 53.73kD and the theoretical pI is 6.12 (#KM520390.1). Phylogenetic tree analysis revealed that gene is related to GmWRKY3. Conserved domains analysis revealed the presence of two conserved WRKY domains and gene belongs to group I WRKY family. Further, MuWRKY3 gene was cloned into plant transformation vector pCAMBIA2301 under the control of CaMV 35S promoter. Subcellular localization studies in tobacco epidermal cells showed the presence of MuWRKY3 in the nucleus. In planta transformation of groundnut carrying MuWRKY3 was standardized using pCAMBIA2301:MuWRKY3 as binary vector. Putative groundnut tranformants were screened on a medium containing 200 mg/L kanamycin. T1 plants were analyzed for molecular and physiological parameters to confirm transgenic groundnut plants. T2 plants demonstrated increased activity of antioxidative efficiency and osmolite accumulation over wild plants.
PA038 (IPPC0305) Identification of high temperature stress-responsive proteins in bread wheat (Triticum aestivum L.) Divya Mishra*, Shubhendu Shekhar, Subhra Chakraborty and Niranjan Chakraborty National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Ambient temperature of earth is drastically increasing due to the effect of global warming. The IPCC report (2014) suggests an approximate increase of 4-5ºC by the end of 21st century. This elevation in temperature has certainly a negative impact on the growth and metabolism of plant, and crop production. Keeping this in mind, we sought to know how plant responds to high temperature stress and the underlying mechanism of tolerance. Bread wheat is one of the most important crop species, with global annual production currently over 600 million tonnes. It is a winter season crop and hypersensitive to high temperature stress. Wheat simulation model predicted that the impact of high temperature stress is more severe than that of dehydration for wheat. Most of the molecular understanding of high temperature stress-responsive cellular adaptation has evolved from transcriptome analyses. However, comparative analysis of high temperature stress-responsive proteins, particularly proteins in the subcellular fraction, is limiting. We screened the status of thermotolerance of several varieties of bread wheat using different physiological and biochemical indexes, and selected relatively tolerant and sensitive cultivars for comparative proteomics analysis. This study would reveal differentially regulated organellar proteins, presumably involved in cellular adaptation and impinge on the molecular mechanism of thermotolerance in wheat. The findings may also provide the basis for targeted alteration of metabolic routes for effective engineering strategies in crop species for agricultural benefits.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 133 Session A : Abiotic Stress Adaptation and Management
PA039 (IPPC0312) Amelioration of salt stress by exogenous application of salicylic acid and arbuscular mycorrhiza in Cicer arietinum L. (chickpea) genotypes Neera Garg and Amrit Bharti * Department of Botany, Panjab University, Chandigarh-160014, India *Presenting author: [email protected]
Soil salinity is one of the major environmental stresses affecting crop production worldwide. Cicer arietinum L. (chickpea) is the third most important food legume grown in the world and is considered highly sensitive to salt stress. Salicylic acid (SA) is a well known phenolic compound and is extensively studied as a growth regulator, since it plays a key role in eliminating the adverse effects of abiotic stresses. In addition, arbuscular mycorrhizal (AM) fungi have also been reported to have a great potential to reduce negative effects of salt stress. Therefore, a study was conducted to investigate the impact of exogenous application of Salicylic acid (0.5 mM) along with arbuscular mycorrhiza on AM colonization, ionic status, growth and yield under various concentrations of NaCl (0, 4, 6, 8 ds m-1) in two chickpea genotypes (salt-tolerant + + + + PBG-5, salt-sensitive BG-256). Salinity negatively affected plant growth, mycorrhizal colonization, K /Na , Ca2 /Na ratios leading to decline in plant productivity. AM inoculation significantly improved root and shoot dry weights, nutrient status (P, K), chl a/b ratio, carotenoid, anthocyanin contents, reduced Na+ uptake and increased ultimate yield. SA enhanced % AM colonization and further improved the growth and productivity when given in combination (SA+AM). PBG-5 displayed better ability to colonize and ensured better nutrient status and growth, which manifested itself in terms of significantly higher seed yield as compared to BG-256. The study concluded that salicylic acid along with arbuscular mycorrhizal inoculation can be applied as a usefull strategy to ameliorate salt stress in chickpea for sustainable agriculture.
PA040 (IPPC0324) Improved drought resistance in a wheat stay green CMS line is associated with enhanced antioxidant competence compared to its fertile line under water deficit stress Renu Khanna-Chopra and Vimal Semwal* Stress Physiology Lab, Water Technology Centre, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Reproductive sink regulates monocarpic senescence in wheat as desinking delays senescence. Wheat fertile (HW 2041) and its isonuclear male sterile line (CMS) were subjected to post-anthesis water deficit stress (WDS) to understand the association between sink strength, oxidative stress, antioxidant defense and drought resistance. CMS plants maintained better water relations and exhibited delayed onset and progression of flag leaf senescence in terms of green leaf area, chlorophyll and protein content than fertile plants. Stay green character in CMS plants under WDS was associated with less ROS generation, lower damage to membranes and better antioxidant defense both in terms of antioxidant enzyme activities and metabolite content compared to fertile plants. CMS plants also exhibited delay in expression of senescence associated genes i.e. WRKY53, serine and cysteine protease (WSP and WCP2), glutamine synthetase (GSI and GSII) and catalase under WDS compared to fertile plants. CMS line exhibited better antioxidant defense in chloroplasts than its fertile line. In conclusion stay green trait contributed to improved drought resistance (DR) in wheat. Reproductive sink regulated drought response as CMS wheat plants exhibited stay green character and improved DR coupled with enhanced antioxidant competence than its fertile line under WDS.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 134 Session A : Abiotic Stress Adaptation and Management
PA041 (IPPC0325) Performance of sugarcane varieties under different moisture stress in subtropical zone Pooja Dhansu1*, A.S. Nandwal1, Mehar Chand2, Ashwani Kumar3 and Anita Kumari4 1Department of Botany & Plant Physiology, CCS Haryana Agricultural University, Hisar-125004, Haryana, India 2Regional Research Station, CCS HAU, Karnal-132001, Haryana, India 3ICAR-Central Soil Salinity Research Institute, Karnal-132001, Haryana, India 4Department of Chemistry & Biochemistry, CCS HAU, Hisar-125004, Haryana, India *Presenting author: [email protected]
A field experiment was conducted at Regional Research Station, CCSHAU, Karnal during 2014-15 with the objective to evaluate the effect of different soil moisture regimes (during pre-monsoon period) on growth and yield of sugarcane varieties. The experiment consisting of three moisture regimes based on available soil moisture (ASM), i.e., 50 % ASM (control), 40% ASM (mild stress) and 30% ASM (severe stress) in main plot and four commercial varieties, i.e., CoS 767, CoH 128 (mid late) and CoJ 64, Co 0238 (early) in sub plot was laid out in split plot design with three replications. The soil moisture stress was started after 45 days of planting (DAP). Significant differences were observed among varieties for germination, growth and yield parameters. Sugarcane variety CoJ64 showed higher germination (64.2%), followed by CoS 767, CoH 128 and lowest in Co 0238. A significant reduction in stalk height (up to 32-43 %), number of tillers (11- 24%), millable canes (up to 24%), cane length, cane growth and cane yield were observed in different varieties under water stress treatment at 30 and 40% ASM as compared to 50% ASM (control). Highest numbers of tillers were recorded in the first week of July. Total chlorophyll content significantly decreased (up to 27%) in stressed plants as compared to control plants. Highest numbers of tillers were recorded in variety CoH 128, followed by CoS 767, CoJ 64 and lowest in Co 0238 in the month of July. Highest numbers of millable canes were recorded in CoS 767, followed by CoH 128, Co 0238, and lowest in CoJ 64.
PA042 (IPPC0328) Differential response of medium duration pigeonpea genotypes to water deficit stress at flowering N. Jyothi Lakshmi*, M. Vanaja, S.K. Yadav, Amol Patil, P. Sathish, Ch. Ram Prasad, Jainender, K. Salini, Arun K. Shanker, B. Sarkar, M. Maheswari and Ch. Srinivasa Rao Central Research Institute for Dryland Agriculture, Santhoshnagar-500059, Hyderabad, India *Presenting author: [email protected]
Four medium duration pigeon pea genotypes (Asha, GT-1, Maruti, Vipula) were evaluated at flowering stage in field under well watered and water deficit stress conditions to assess the effect of water-deficit stress and its relief on physiological and biochemical parameters including the assay of antioxidant enzymes to ascertain their role in stress tolerance. Water deficit reduced the leaf area per plant in all genotypes. The percent reduction in seed yield with water deficit stress was 46.2, 57.4, 50.4, 54.3% and drought tolerance efficiency (DTE) was 53.8, 42.6, 49.5, and 45.7 for genotypes Asha, GT- 1, Maruthi and Vipula respectively. The drought susceptible index (DSI) was 0.90, 1.11, 0.97 and 1.05 for genotypes Asha, GT-1, Maruthi and Vipula respectively. Genotype Asha was found to be less sensitive to moisture stress condition for days to 50% flowering as well as maturity. The genotype Asha was found to be promising for total dry matter production under moisture stress (61.8 g/plant) as well as irrigated (115.2 g/plant) conditions. Genotype Asha followed by Maruti were superior over Vipula and GT-1 in terms of leaf water relations, accumulation of compatible solutes, photosynthesis, WUE, membrane stability index and activity of antioxidant enzyme viz., superoxide dismutase (SOD). Genotype Asha recorded highest grain yield and total dry matter, low reduction in yield due to water stress, high DTE, low DSI, less reduction in HI under stress. Maintenance of superiority in terms of these physiological and biochemical parameters coupled with better recovery ability upon stress relief are crucial physiological mechanisms contributing to water deficit stress tolerance in the genotype Asha.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 135 Session A : Abiotic Stress Adaptation and Management
PA043 (IPPC0343) Over-expression of BrHsf3 helps in multiple stress tolerance in Brassica juncea (Indian mustard) Subhash Chandra Prasad1*, Philip Mullineaux2, Ulrike Bechtold2 and Neera Bhalla Sarin1 1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2University of Essex, UK *Presenting author: [email protected] Heat-shock transcription factors (HSFs) are known to play an important role in regulation of expression of heat shock proteins in eukaryotes. Plants have a much higher number of HSFs in comparison to other species and each HSFs have unique functions in response to environmental stress. Earlier studies indicated that HSF3 acts beyond its previously reported function. It seems to stand at the interface of abiotic and biotic stress signalling pathways and thus HSF3 overexpression confers multiple abiotic and biotic stress resistance. Brassica species are highly sensitive to heat stress at the flowering stage that causes a decline in seed yield. In the present study Brassica juncea (Indian mustard) was genetically engineered by overexpressing BrHsf3 (Brassica rapa; Br, Heat shock factor 3) gene and the morphological and physiological characters of the transgenic lines were compared with the untransformed control plants and it was found that different transgenic lines showed normal morphology and growth similar to the untransformed (UT) control plants. Leaf disc senescence assays undertaken for two representative transgenic lines showed increased tolerance when exposed to NaCl and mannitol stress. Histochemical staining of transgenic lines exposed to heat stress (35°C for - 12 hr) showed lower accumulation of hydrogen peroxide (H2O2) and superoxide (O2 ) radicals in leaves. The specific enzyme activity of ascorbate peroxidise (APX) was increased in transgenic lines in comparison to UT under heat stress condition this could be the reason, less ROS accumulating in transgenic lines in comparison to UT.
PA044 (IPPC0348) Rice nuclear protein OsNAPL6 is an H3/H4 histone chaperone functioning in response and adaptation to various abiotic stresses Amit Tripathi1*, Ashwani Pareek2 and Sneh Singla-Pareek1 1Plant Molecular Biology Group, Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India 2Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected] Understanding the mechanistic details of stress response in plants is a prerequisite to develop superior crop varieties that may withstand environmental extremes. Focusing on ‘master regulators’ which can drive the expression of a variety of downstream target genes is considered to be a significant approach in this endeavour. By altering the DNA accessibility, histone chaperones represent an important class of such ‘master regulators’; however, their physiological role in plants vis-à-vis abiotic stresses largely remains enigmatic. We have previously found significant differential regulation of a few histone chaperones under one or more environmental stress conditions. In the present study, we narrowed down to a stress-responsive member of the NAP superfamily of histone chaperones, OsNAPL6, which is expressed at a higher level in the salt-tolerant genotype Pokkali under most of the abiotic stress conditions. We found OsNAPL6 to be a nuclear- localized H3/H4 histone chaperone capable of assembling nucleosome-like structure on a relaxed plasmid template. Further, we utilized overexpression and knockdown approach to carry out detailed functional characterization of OsNAPL6 with respect to its role in abiotic stresses in rice. We showed that alteration in cellular levels of OsNAPL6 positively correlates with adaptation to various abiotic stresses as measured via analyzing morphological, physiological and agronomic traits. Considering that stress response and adaptation require altered expression of thousands of genes, together, our results suggest that the stress-responsive histone chaperone OsNAPL6 may serve a regulatory role in stress physiology by controlling the expression of stress-related genes, possibly via modulating nucleosome dynamics at the corresponding loci.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 136 Session A : Abiotic Stress Adaptation and Management
PA045 (IPPC0356) Effects of water stress on crop physiology of groundnut, aerobic rice, soybean and management options for mitigation K.G. Mandal1*, A.K. Thakur1 and K. Kannan2 1ICAR-Indian Institute of Water Management (Formerly Directorate of Water Management) Bhubaneswar-751023, Odisha, India 2Regional Research Station, ICAR-Indian Institute of Soil and Water Conservation, Ooty, Tamil Nadu, India *Presenting author: [email protected]
Field experiments were conducted on rice and groundnut, and a pot experiment on soybean to study water stress effects on crop physiology under different management. For groundnut, fv/fm and actual fluorescence efficiency (ÔPS II) showed lower values under water stress compared to well-watered plants; intercepted PAR and pod yield was greater (64.5- 74.4%) with paired-row planting on beds than flat-bed (46.3-57.7%); hence, paired-row planting of summer groundnut with optimum irrigation to its critical growth stages would mitigate crop water stress and enhance WUE by 39%. Water stress effect on aerobic rice was prominent on cross section of xylem vessels and cortex cells, root mass density, fv/fm, ÖPS II, yield attributes and grain yield than transplanted rice; the varieties viz. ‘Surendra’, ‘Apo’ and ‘Lalat’ showed the highest yield potential (3.9-4.6 t ha-1) under aerobic conditions. Results on pot experiment on soybean showed physiological changes in soybean seedlings. Due to water stress, relative leaf water content, SPAD values, total chlorophyll, rate of photosynthesis, fv/fm, ÔPS II and soluble protein content decreased, whereas proline content and enzyme activities i.e. catalase (CAT), superoxide dismutase (SOD), guaiacol peroxidase (POD) activity and malondialdehyde (MDA) content increased in soybean seedlings due to increase in reactive oxygen species (ROS). However, application of methyl jasmonate (MeJA @ 1 mmol L-1) reduced the adverse effects due to ROS and thereby increased rate of photosynthesis, stomatal conductance and soluble protein content. Hence, methyl jasmonate has the potential to mitigate water stress and to enhance the water stress tolerance to soybean seedlings.
PA046 (IPPC0361) PROTEIN L-ISOASPARTYL METHYLTRANSFERASE confers tolerance to abiotic stress in Arabidopsis thaliana Shraboni Ghosh*, Pooja Verma and Manoj Majee National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Abiotic stresses usually lead to deterioration of structure and function of proteins. Maintenance of such proteins during stress condition is vital for survival under stress. PROTEIN L-ISOASPARTYL METHYLTRANSFERASE (PIMT, EC.1.1.77) is a protein repairing enzyme that is known to repair damaged and aged proteins. Here we examine the function of PIMT in Arabidopsis thaliana plants under salinity, dehydration, oxidative, heat and cold stress conditions. Involvement of PIMT in stress tolerance was investigated by analysis of PIMT overexpression and RNAi lines in seedlings as well as in mature plants. During stress conditions elevated PIMT activity was seen which limits the accumulation of abnormal L-isoAsp residues. The RNAi lines exhibited greater sensitivity to the stress conditions whereas constitutive overexpression of both cytoplasmic (AtPIMT1) and nuclear (AtPIMT2) PIMT confers tolerance to various stresses. The damage caused by induced stresses were assayed in terms of malondialdehyde content, chlorophyll estimation and H2O2 accumulation. The overall higher PIMT activity and lower isoaspartyl accumulation leads to improved survival of the transgenic plants. The results indicate that PIMT not only functions in seed vigor and longevity but also provides stress tolerance to plants by repairing isoAsp mediated damaged proteins which in turn limit reactive oxygen species formation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 137 Session A : Abiotic Stress Adaptation and Management
PA047 (IPPC0366) Ectopic expression of a wheat SNARE-binding protein confers drought tolerance in transgenic Arabidopsis thaliana Brinderjit Singh1*, Paramjit Khurana1, J.P. Khurana1 and Prabhjeet Singh2 1Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India 2Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab 143005, India *Presenting author: [email protected]
Drought stress is one of the major factors which limits crop productivity and has been reported to result in upto 50% loss in the average grain yield. The response of plants to drought stress is highly complex and is contributed by multiple genes. Therefore, identification of genes involved in stress tolerance in crops is a prerequisite for developing stress tolerant plants by transgenic approaches. In the present study, gene encoding for vesicle-associated membrane protein (VAMP)-associated protein (TaVAP) was used to develop transgenic Arabidopsis plants for studying its role in abiotic stress tolerance. The VAMP-associated proteins (VAPs) have been proposed to play roles in vesicle trafficking, microtubule organization, lipid metabolism, inositol auxotrophy and unfolded protein response. However, the physiological role of these genes is still a matter of conjecture in plants. This study demonstrated that VAP is functionally conserved, since TaVAP was able to rescue the yeast mutant cells, which were mutated in orthologue of this gene. Our studies provide the first evidence that TaVAP play an important role in abiotic stress response in plants. The expression of this gene was enhanced in wheat seedlings in response to different stress conditions. Furthermore, ectopic expression of TaVAP in Arabidopsis conferred significantly higher tolerance to transgenic plants under abiotic stress conditions. Importantly, the overexpression of TaVAP did not result in any yield penalty, thereby, indicating that this gene is an ideal candidate for enhancing abiotic stress tolerance of crops through transgenic approaches.
PA048 (IPPC0369) Differentially expressed galactinol synthase gene (s) of chickpea (Cicer arietinum L.) improves seed vigor and longevity by limiting the hydroxyl radicals during aging Prafull Salvi* and Manoj Majee Plant Molecular Biology and biochemistry, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected] Galactinol synthase (EC: 2.4.1.123) (GolS) catalyzes the rate limiting step in Raffinose Family Oligosaccharide (RFO) biosynthetic pathway. RFOs including galactinol are known as anti-stress agents and suggested to play important role in abiotic stress responses and seed desiccation tolerance. In this study, we have shown galactinol synthase activity as well as galactinol content is highly concentrated in pod and dry seed and gradually decreased as seed germination progresses. Galactinol synthase activity was also found to be enhanced in response to stresses. In chickpea, galactinol synthase enzyme is encoded by two divergent genes (CaGolS 1 and CaGolS 2) which produce 5 different types of transcripts. Transcription analysis revealed that the CaGolS1 and CaGolS2 are differentially regulated. Further, all isoforms were bacterially expressed, purified and biochemically characterized. All isoforms exhibit differential enzyme activity. Native molecular weight and subunit composition was determined for enzymatically active isoform through gel filtration chromatography. Confocal microscopy showed that a transiently transfected CaGolS-YFP fusion proteins primarily distributed to nucleus and membrane in onion epidermal cell. Further we found that seed-specific expression of CaGolS1 and 2 in Arabidopsis play a role in maintaining seed vigor and longevity, which was correlated well with increased galactinol and raffinose contents and reduced hydroxyl radicals in seed of transgenic lines. Collectively, our work clearly demonstrates that CaGolS (s) are differentially regulated in chickpea and play important role in enhancement of seed vigor and longevity.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 138 Session A : Abiotic Stress Adaptation and Management
PA049 (IPPC0371) Surviving metabolic arrest: understanding the mechanisms of desiccation tolerance in the resurrection plant Craterostigma plantagineum Dinakar Challabathula* Department of Life Sciences, Central University of Tamil Nadu, Thiruvarur-610101, Tamil Nadu, India *Presenting author: [email protected]
Drought is the most common abiotic stress that is encountered by the plants. Most angiospermic plants are drought sensitive and do not survive if the relative water content decreases below a certain limit, whereas, the seeds of many plants can tolerate extreme dehydration. A small group of resurrection plants from Linderniaceae family have developed the ability to survive severe water loss in vegetative tissues and thus regarded as desiccation tolerant eg. Craterostigma plantagineum. Transcriptome analysis of C. plantagineum at different dehydration stages identified the genes that are up-regulated/down-regulated during desiccation. Existence of both desiccation tolerant and sensitive plants in Linderniaceae family provides an advantage for comparative analysis there by to understand the mechanisms of desiccation tolerance. Transcripts encoding group II late embryogenesis abundant (LEA) proteins from C. plantagineum were identified and quantified. While the transcripts encoding LEA-like 11-24 protein are abundantly expressed during desiccation in the desiccation tolerant plants C. plantagineum and Lindernia brevidens, the transcripts are expressed at a low level in the desiccation sensitive Lindernia subracemosa. Differential activities of the promoters were observed along with some conservation in Cis elements. While desiccation sensitive plants showed degradation of mRNA of specific genes during desiccation, in the desiccation tolerant plants the mRNA for these genes was highly stable. Further, comparative studies utilising physiological, genetic and biochemical approaches are essential to decipher the underlying mechanisms of desiccation tolerance.
PA050 (IPPC0389) Effects of gamma radiation stress on production of 20-hydroxyecdysone in Sesuvium portulacastrum L. shoot cultures Vishwadeep Kapare1*, Ramesh Satdive2, Nutan Malpathak1 and Devanand P. Fulzele2 1Department of Botany, Savitribai Phule Pune University, Pune-411007, India 2Plant Biotechnology and Secondary Metabolites Section, NABTD, Bhabha Atomic Research Centre, Mumbai-400085, India *Presenting author: [email protected]
Sesuvium portulacastrum Linn. a member of family Aizoaceae, is naturally growing plant in coastal areas and rich source of phytoecdysteroid 20-hydroxyecdysone (20E). Various biological properties including antidepressant, antioxidant, prevention of infectious diseases and cytotoxic activity are associated with 20E. Due to its anabolic activities it is commonly used by body builders and sportsmen in form of phytoecdysteroid containing herbal extracts as well as OTC drug. In vitro shoots cultures of S. portulacastrum were irradiated with low doses of 5, 10, 15, 20, 25, 30, 35 and 40 Gy from a cobalt- 60 source. Growth kinetics and content of 20E of the in vitro cultures were studied for three generations: M1, M2, and M3. High Performance Liquid Chromatography (HPLC) revealed that gradual increase in gamma doses resulted in significant increase in product synthesis. Shoot cultures were irradiated at 20 Gy synthesized highest 1.19 mg/gm of 20E, whereas non irradiated shoot cultures produced 0.35 mg/gm. High doses of gamma rays impaired product synthesis and growth rate of shoot cultures. The present investigation corroborated that in vitro irradiation is an important method for improvement of secondary metabolites at suitable doses of gamma rays.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 139 Session A : Abiotic Stress Adaptation and Management
PA051 (IPPC0403) Physiological trait based breeding: An obligate for future food security under abiotic stress H.M. Mamrutha1*, Ritika Choudhary1, Dheeraj Rana1, Omprakash Tuteja1, Ravish Chatrath1, Sai Prasad2, D.C. Saxena2, Satish Mishra3, Sonia Sheoran1, Chetan Kumar Choudhary1, Mitchelle Watt4 and Indu Sharma1 1Indian Institute of Wheat and Barley Research, Karnal-132001, Haryana, India; 2Indian Agricultural Research Institute (IARI) Regional Station, Indore, India; 3Agarkar Research Institute, Pune-411004, India; 4CSIRO, Australia *Presenting author: [email protected] By the year 2050, the projected global and India’s wheat demand will be 1090 and 140 mt, respectively. This target can be achieved with annual production growth of 1.7%. There is ceiling in wheat yield under high productive areas, hence the other potential source to increase yield is stressed environment. Worldwide 84 mha of wheat growing area is affected by drought, and traditional breeding efforts are focused on yield rather than survival, which present a challenge to wheat scientists to develop wheat cultivars that are both productive and adapted to abiotic stresses. Recently physiological traits (PTs) have become a target for plant breeders aiming to improve yield potential in stressed environment. In the present study, the different PTs, viz., early vigour, tillering inhibition, deep rooting system and alternate dwarfing genes for deep sowing under drought were studied. A set of wheat genotypes were validated using novel field screening methods in three different locations in India and Australia. Morphological markers and germplasm source for these traits have been identified. The morphological marker includes third leaf width at seedling stage, as a proxy for early vigour, long coleoptile length for emergence from deeper soil, spike pubiscence for tillering inhibition and canopy temperature at 15 days after anthesis for deep root system. The germplasm identified were MW257, Synthetic 9589/Kennedy, CM18/Viguor18 with Rht13 and HI 1531 for tillering inhibition, early viguor, long coleoptile length and deep root system, respectively. These traits have been introgressed into Indian wheat genotypes, mapping populations have been developed and identification of relevant molecular marker is under progress.
PA052 (IPPC0404) Differential transcript abundance of genes of antioxidative enzymes and redox cues reveal two faces of hydrogen peroxide-mediated oxidative stress in rice seedling Chakrabarty Ananya* and Bhattacharjee Soumen Centre for Advanced Study, Department of Botany, The University of Burdwan, Burdwan-713104, West Bengal, India *Presenting author: [email protected]
Treatments with different titre of H2O2, both elevated (20 mM) and low (500 μM) during early imbibitional phase of germination resulted in the generation of differential redox cues in metabolic interface as being investigated in terms of significant changes in prooxidant/antioxidant ratio, titre of individual reactive oxygen species, efficacy of Ascorbate-Glutathione cycle, activities of enzymatic antioxidants, radical scavenging activities, changes in thiol compounds and expression of oxidative damage to membrane protein and lipid. All the redox parameters tested, corroborate the finding that treatment with elevated concentration of H2O2 cause serious loss of redox homeostasis, whereas low titre H2O2 treatment not only restores redox homeostasis but also up-regulate oxidative stress-acclimatory processes as evident from reduction in oxidative damages of newly assembled membrane system. 500 μM H2O2 treatment exhibited enhanced activities of Ascorbate-Glutathione pathway enzymes, which was otherwise inhibited significantly in 20 mM treated seedlings. A comparison between imbibitional chilling stress-raised experimental rice seedling with 20 mM H2O2 treated rice seedling reveal similar kind of generation of redox cues. Further, H2O2 treatments in rice seedlings also reveal the dose-dependent regulation of expression of genes of antioxidative enzymes. Low titre H2O2 treatment combat oxidative stress by up- regulation of gene expression of some antioxidative enzymes, particularly of Ascorbate-Glutathione pathway, which in turn offers better capacity of redox regulation and mitigation of oxidative damage of juvenile membrane system. In conclusion, a dose-dependent regulation of H2O2 mediated redox cues, redox regulatory properties and oxidative damages in rice seedling is suggested.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 140 Session A : Abiotic Stress Adaptation and Management
PA053 (IPPC0405) Efficacy of expression of genes of ascorbate-glutathione cycle and redox regulatory mechanisms under dehydration stress as the basis of screening land races of some aromatic rice of West Bengal D. Nivedita* and Bhattacharjee Soumen Centre for Advanced Study, Department of Botany, The University of Burdwan, Burdwan-713104, West Bengal, India *Presenting author: [email protected] PEG-induced dehydration stress during early germination causes disruption of redox-homeostasis and oxidative damage in four aromatic land races (Jamainadu, Tulaipanji, Sitabhog and Badshabhog) by enhancing the accumulation of pro- .- oxidants (DCFDA oxidation, O2 and H2O2 accumulation), significant reduction of antioxidative defence (total antioxidant and radical scavenging capacity, total thiol content and activities of antioxidative defence enzymes) and aggravating protein oxidation and membrane lipid peroxidation (assessed in terms of free carbonyl content and accumulation of thiobarbituric acid reactive substances). When compared between the land races, a clear trend in differential redox regulatory properties in which ROS-antioxidant interaction acts at metabolic interface was observed in the order of the land races Tulaipanji> Sitabhog> Badshabhog> Jamainadu. Moreover, when the efficacy of Ascorbate- Glutathione cycle for scavenging H2O2 generated during dehydration stress were assessed and compared between the land races exposed to PEG-induced dehydration stress, it also exhibited almost the same trend with the land race Tulaipanji and Jamainadu exhibiting maximum and minimum efficiencies respectively. The land race Tulaipanji and Sitabhog resist dehydration stress better than the other land races due to its early preparedness to combat oxidative stress by upregulating expression of genes of some enzymes of Ascorbate-Glutathione cycle along with some other antioxidative enzymes. A model of redox homeostasis in which ROS- antioxidant (Ascorbate- Glutathione system) acts at metabolic interface for up-regulation of antioxidative gene expression necessary for differential drought stress tolerance among the land races of aromatic rice is suggested.
PA054 (IPPC0407) Loss of adaptive plasticity of mangroves in degraded ecosystem-a study based on osmolyte accumulation Krishna Ray1*, Sandip Kumar Basak2, Tapan Sutradhar1, Chandan Mukherjee1, Rajojit Chowdhury1 and Momtaj Begam1 1Department of Botany, Environmental Biotechnology Group, West Bengal State University, Barasat-700126, West Bengal, India 2Department of Botany, Kandi Raj College, Kandi, Murshidabad, West Bengal, India *Presenting author: [email protected] In degraded mangrove forests, it has been well documented that due to various environmental constraints, the ecosystem homeostasis gets hindered to an extent when it can no longer continue the normal processes of secondary succession and mangrove forests come to the verge of extinction. Mangroves bear a fascinating complex network of morpho- physiological adaptations, but their loss of adaptive plasticity is causing gradual disappearance under degraded ecosystem. Synthesis of nitrogen-rich and energy-intensive organic solutes proved to be strong selective advantage for mangroves to encounter salt stress, but this adaptation is too expensive in terms of metabolic costs for this group of plants resulting into inevitable trade-off against its growth. The cost of generating enough organic solutes to achieve full osmotic adjustment in hypersaline soil exceeds all other energy consuming adaptive processes. It becomes far more challenging for mangroves, when they have to adjust osmotically under degraded soil conditions. Our studies established that nutrient cycling under degraded mangrove soil is largely impaired and limit the synthesis of nitrogen-rich and reduced carbon based compatible solutes. The pioneer colonizer halophytic grasses like Porteresia, Myriostachya, Paspalum, and Sporobolus sp. In Sundarban mangrove ecosystem contribute to nutrient cycling through the bacterial population harbouring at therhizospheric regions, hence augmenting the secondary succession and restoration process in the degraded mudflats. These grasses with their ideal osmotic and strong mechanical adaptivitypave the way for stabilization and stress management of incoming mangrove species under a multitude of abiotic stressesin degraded mangrove ecosystem of Sundarban.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 141 Session A : Abiotic Stress Adaptation and Management
PA055 (IPPC0411) Selection of castor (Ricinus communis L.) germplasm with good root traits for drought tolerance in field P. Lakshmamma*, Lakshmi Prayaga and K. Alivelu Department of Plant Physiology, ICAR-Indian Institute of Oilseeds Research, Rajendranagar- 500030, Hyderabad, India *Presenting author: [email protected]
Fourteen germplasm lines with good and two with poor root growth identified by screening in root structures along with a variety (48-1) and hybrid (DCH-519) check were sown in field for two years during late rabi (Nov. 1st week) 2010-11 & 2011-12 and water stress was imposed from 30-90 days after sowing (DAS) by withholding irrigation in three replications in split plot design. Pooled analysis of data shows significant growth reduction in terms of reduced plant height, leaf number, stem girth, branches and total dry matter (56%) with water stress. On an average, there was 13% reduction in TDM at harvest, 35% reduction in seed yield and 19% reduction in harvest index (HI) with drought stress. Among the genotypes studied, RG 1494 and RG 2139 recorded <20% reduction in seed yield and also showed low drought susceptibility index (DSI) of <0.5. Primary seed yield reduced significantly with stress, but the reduction was less in secondaries and compensated in later order spikes in these genotypes. The root growth in terms of root volume (RG 1494-310 cm3, RG 2139-557 cm3) and root dry weight (RG 1494-122 g/pl., RG 2139-93 g/pl.) was also more in these genotypes. Chlorophyll content measured with SPAD chlorophyll meter (SCMR) and bloom (wax) content increased in stressed plants as an adaptive feature. The genotypes RG 1494, RG 2139 that showed good performance during both years with good root and shoot growth characters can be used in drought breeding programs.
PA056 (IPPC0413) Differential accumulation and gene expression of various osmolytes in Brassica juncea under temperature stress Sudhir Kumar1*, R.K. Sairam1, K.V. Prabhu2 and Akshay Sakhare1 1Division of Plant Physiology, 2Division of Genetics, Indian Agriculture Research Institute, New Delhi-110012, India *Presenting author: [email protected]
A study was carried out to evaluate the role of various osmolytes in temperature stress tolerance in Brassica juncea genotypes. Three dates of sowing were used for temperature stress i.e. D1 (30 Oct), D2(15 Nov) and D3(30 Nov). Different osmolytes were studied. Result showed that Temperature stress due to late sowing (D2 and D3) resulted in increase in contents of total sugar, proline, glycine-betaine and trehalose as compared to normal (D1) sowing. Among various genotypes CS 52, Progaro and NDR 8801 accumulated highest content of different osmolytes as compared to Pusa Agrani, EJ-15 and Pusa Tarak, which showed lesser accumulation under temperature stress. Gene expression studied by RT-PCR showed temperature stress induced increase in mRNA transcript of β-pyrroline-5-carboxylate synthetase (P5CS), betaine aldehyde dehydrogenase (BADH) and trehalose-6 -phosphate synthase (T6PS), which was again higher in Proagro and CS 52 compared to Pusa Agrani, EJ 15 and Pusa Tarak. Results suggested that high temperature induced expression of P5CS, BADH and T6PS in Proagro and CS 52 resulted in greater accumulation of osmolytes, which could be one of the reasons of their tolerance to temperature stress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 142 Session A : Abiotic Stress Adaptation and Management
PA057 (IPPC0418) Evaluation of rice varieties and hybrids under high temperature stress Beena Radha1*, Veena Vigneswaran1, P. Sindhumol1, M.C. Narayankutty1 and S.R. Voleti2 1Department of Plant Physiology, Regional Agricultural Research Station, Pattambi, Kerala Agricultural University, Palakkad-679306, Kerala, India 2Department of Plant Physiology, Directorate of Rice Research, Hyderabad-500030, AP, India *Presenting author: [email protected]
A study was conducted to evaluate the performance of rice varieties and hybrids under high temperature stress during 2014 at Regional Agricultural Research Station, Pattambi, Kerala Agricultural University. In this study seven rice hybrids and seventeen hybrid derivatives were evaluated at 6-8°C above the normal temperature and ambient condition from seedling to maturity stage. Biomass accumulation of high temperature exposed plants were higher than plants kept at ambient condition. Spikelet fertility of rice hybrids were very low but hybrid derivatives were maintained spikelet fertility as similar to that of ambient condition. In another study it is revealed that when plants were shifted to high temperature from panicle initiation to maturity stage, hybrid derivatives also showed reduction in spikelet fertility. From these studies, it can be concluded that when plants were exposed to high temperature from seedling stage to maturity occurrence of spikelet sterility is very less due to the acclimatization process but hybrids were highly susceptible to high temperature stress than hybrid derivatives. Spikelet fertility of hybrid derivatives ranged from 78.35% (Lalat) under high temperature and 92.70% (IET 23216) under ambient condition. Spikelet fertility of rice hybrids ranged from 64.52% (IET 24122) under high temperature and 90.22% (IET 24117) under ambient condition. Photosynthetic rate and spikelet fertility of high yielding varieties were maintained on par between ambient and high temperature condition except Lalat and MTU-1010. In rice hybrids, both photosynthetic rate and spikelet fertility significantly reduced under high temperature condition except in IET 21404 and US-312.
PA058 (IPPC0425) Characterization of delayed senescence traits in chickpea (Cicer arietinum L.) germplasm under drought stress Gurumurthy Shekharappa1*, P.S. Basu1, Arora Ajay2, Jagdish Singh1 and N.P. Singh1 1Division of Crop Physiology, Biochemistry & Microbiology, Indian Institute of Pulses Research, Kanpur-208024, India 2Division of Plant Physiology, ICAR-Indian Agriculture Research Institute, New Delhi-110012, India *Presenting author: [email protected]
An experiment was conducted with 276 chickpea genotypes of ICRISAT minicore with an objective to screen the genotypes having drought induced delayed leaf senescence trait. The physiological parameters such as SPAD chlorophyll meter reading (SCMR), phenology, water use efficiency (WUE), membrane stability index (MSI), leaf Senescence Rate (LSR), greenness index (GI), yield and yield components were studied. We identified thirteen contrasting genotypes belong to delayed senescence and the other thirteen falls in the category of faster senescence among the genotypes studied. Higher and lower reduction in the WUE, MSI and SCMR during stress condition was observed with faster and delayed senescence genotypes, respectively under moisture deficit stress condition. These contrasting genotypes did not differed in flowering and podding behavior of both the irrigated and stress condition. Delayed senescence genotypes showed lesser reduction in leaf LSR and significantly higher GI compared to genotypes with faster senescence. Genotypes with delayed senescence showed higher biomass as well as better adaptation under drought. LSR was recorded by visual scoring; the maximum and minimum score was recorded in genotypes ICC 762 and ICC 6279, respectively. The present study revealed that SCMR, WUE, MSI, LSR and GI are good physiological indices under water deficit stress condition for chickpea crop and can be used for screening of chickpea genotypes for drought induced leaf senescence.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 143 Session A : Abiotic Stress Adaptation and Management
PA059 (IPPC0427) Assessment of terminal heat tolerance ability of wheat genotypes based on physiological traits using multivariate analysis Shivani Nagar1*, V.P. Singh1, Ajay Arora1, Neera Singh3, Rajkumar Dhakar2 and Shiv Ramakrishnan1 1Division of Plant Physiology, 2Division of Agricultural Physics, 3Division of Agricultural Chemicals, Indian Agricultural Research Institute, New Delhi, India-110012, India *Presenters author: [email protected]
Terminal heat stress is one of the limiting factors in wheat production and it is expected to rise under present climate change scenario. Present study was conducted to evaluate the performance of 40 wheat genotypes under terminal heat stress conditions based on eight physiological traits. The plants were sown late (i.e., on 5th January) to expose them to terminal heat stress. The genotypes were evaluated using multivariate analysis, viz., Ward’s method of hierarchical clusters analysis, discriminant analysis and principle component analysis. The genotypes were categorized in to three groups namely tolerant, intermediate and sensitive. Tolerant genotypes like DBW-14, RAJ-3765, HD-2643 and HALNA performed physiologically better in terms of higher membrane stability index (MSI), chlorophyll content, photosynthesis rate (PN), harvest index (HI) under heat stress condition. Genotypes HD-2987, SHAINGHAI-1, HD-2402 and WH-730 were found to be heat sensitive. Physiological traits like MSI, SPAD value, Fv/Fm ratio and PN were found to be most important contributor in grouping of genotypes, and showed positive correlations (r) of 0.73, 0.47, 0.41 and 0.39 with grain yield, respectively, which were significant at p<0.05. The large genetic diversity was found among the genotypes based on physiological traits. These genotypes can be utilized in wheat improvement programs for heat tolerance.
PA060 (IPPC0430) Effect of salicylic acid to mitigate the high temperature stress in Indian mustard [Brassica juncea (L.) Czern & Coss B.L. Kakralya* and O.P. Godara Department of Plant Physiology, S.K.N. College of Agriculture, Jobner, Rajasthan, India *Presenting author: [email protected]
A field experiment was conducted to evaluate the role of salicylic acid in mitigating high temperature stress effects in two genotypes of Indian mustard viz.RGN-236 and RGN-229. Both the genotypes were exposed to high temperature through delayed sowing and three concentrations of salicylic acid viz. 50, 100 and 150 ppm were sprayed at pre anthesis and siliqua initiation stage. Under delayed sowing, relative water content, photosynthetic rate, chlorophyll stability index, membrane stability index and oil content decreased significantly. Whereas, transpiration rate, stomatal conductance and leaf temperature significantly increased under delayed sowing. On the other hand, application of salicylic acid showed less reduction in growth and above physiological traits. Among three concentration of salicylic acid, 150 ppm showed the best response and was followed by 100 and 50 ppm. Yield attributes like number of siliqua per plant, number of seed per siliqua, plant height, test weight, seed yield, biological yield, harvest index decreased with delayed sowing under control but plants treated with 150 ppm salicylic acid showed significant improvement and less reductions due to high temperature stress under delayed sowing. Among two genotypes, RGN-229 showed less reductions in growth and yield and can be considered as superior in terms of high temperature tolerance over RGN-236.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 144 Session A : Abiotic Stress Adaptation and Management
PA061 (IPPC0436) Differential response of rice cultivars to high night temperature Sangeeta Khetarpal1, Anjali Anand1*, Archana Yadav2, S.M. Trivedi1, Dinesh Kumar3 and Himanshu Pathak2 1Division of Plant Physiology, 2CESCRA, 3Division of Agronomy, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Global climate change is leading to asymmetric atmospheric warming with reduced temperature differences between day and night. Minimum night temperatures are increasing at a faster pace than maximum day temperatures and this increase is expected to be larger in the tropics and subtropics (IPCC, 2013) which correspond to the main rice production area. Twenty rice cultivars were subjected to high night temperature (HNT) from panicle emergence to maturity. During the exposure period and despite some variability, mean minimum temperature increased by about 1.5oC in the polychambers over the mean minimum temperature of 24oC in ambient. Flag leaf night respiration rates increased manifold in all cultivars accompanied by reduction in biomass. Based on grain yield heat susceptiblity index Nerica L 44, Nagina 22, Ganga and Abhishek performed better under HNT. Leaf starch content reduced by 14% in genotypes showing better performance under HNT compared to 26% in genotypes with high susceptibility index suggesting more loss of stored carbohydrate under HNT. The reduction in stem starch and increase in reducing sugar content observed in most of the genotypes could be due to reduced synthesis of stem starch as a consequence of lower availability of photoassimilates from leaves due to respiratory losses or increased breakdown of starch in stem. Grain filling was adversely affected in the middle stage of grain growth that could be another cause for lower yields under HNT in some genotypes. Quality was affected in terms of decrease in proportion of high density grains and increase in chalkiness and grain protein content under HNT.
PA062 (IPPC0437) Oxidative stress and antioxidative defence system in Ricinus communis (L.) under high temperature stress Praduman Yadav* and C. Lavanya Department of Biochemistry, ICAR-Indian Institute of Oilseeds Research, Rajendranagar, Hyderabad-500030, India *Presenting author: [email protected]
The study was undertaken to investigate the effect of high temperature (HT) induced stress and revival on antioxidative enzymes and metabolites in leaves of the castor (Ricinus communis L.) seedlings of HT susceptible and HT tolerant genotypes. Seeds of thirty five genotypes were sown in trays having sandy loam soil. Ten days old seedlings were exposed to threshold high temperature (45±0.5°C) and 30% relative humidity continuously. Time taken to 50% mortality was recorded in all the genotypes for screening against high temperature tolerance. Two genotypes were identified as HT susceptible (DCS-89 and DCS-117) and two genotypes as HT tolerant (DCS-107 and DCS-113). After subjecting the seedlings to HT stress, the seedlings were revived by placing the trays at 25±0.5°C for 24 hrs. Activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX) and glutathione reductase (GR) increased in both HT tolerant and susceptible genotypes under stress but increase was found significantly higher in tolerant genotypes. On revival antioxidative enzymes viz. SOD and CAT started decreasing, but the activities of POX,
APX and GR still continued to increase in all the genotypes. The malondialdehyde (MDA) content H2O2 concentration and lipoxygenase (LOX) activity in leaves of castor seedlings increased at the high temperature but higher percent increase was observed in susceptible genotypes. However, on revival, LOX activity, MDA and H2O2 content decreased as compared to stressed conditions in all the genotypes.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 145 Session A : Abiotic Stress Adaptation and Management
PA063 (IPPC0446) Overexpression of ABA receptor gene OsABAR6 enhances drought tolerance of rice V.V. Santosh Kumar1*, Shashank K. Yadav1, Sanya Shrivastava1, Rakesh K. Verma1, Suchitra Puskar1, M.V. Rao2 and Viswanathan Chinnusamy1 1Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India Department of Plant Sciences, Bharathidasan University Tiruchirappalli, Tamil Nadu, India *Presenting author: [email protected]
Abscisic Acid (ABA) plays crucial roles in development and stress responses of plants. ABA receptor (ABAR) family in Arabidopsis consists of 14 members. Bioinformatic analyses led to the identification of 12 PYR/PYL homolog ABARs from rice. However, the specific function of members of ABARs in developmental and stress responses needs to be understood. In present study, OsABAR6 gene from rice was used for functional validation. Expression analysis of ABAR6 in different tissues and stress conditions revealed differential expression patterns. ABAR6 gene was cloned in binary vector pCAMBIA1300 under AtRD29A promoter and pANDA RNAi vectors. Trangenic rice plants were developed by using Agrobacterium mediated genetic transformation and confirmed through PCR, qRT-PCR and southern blotting. Rice transgenics expressing AtRD29A::OsABAR6 showed hypersensitivity to ABA in germination as compared with non-transformed (NT) wild type and RNAi lines. This suggests that ABAR6 conferred enhanced ABA perception and thus function as ABA receptor. Analysis of drought tolerance of RD29A:ABAR6 rice transgenics (T2) in pot culture under greenhouse conditions revealed that RD29A:ABAR6 transgenic rice lines are more tolerant than NT plants in terms of maintenance of RWC, membrane stability index, chlorophyll content, survival and recovery growth. However, due to leaky expression of AtRD29A::ABAR6 even under control growth conditions, ABAR6 plants showed reduced height and yield. Water utilization experiment revealed that transgenic plants used about 40% less water per a day as compared to wild type plants. Results from this study suggest that ABAR6 has potential to improve WUE and drought tolerance of rice.
PA064 (IPPC0449) Abiotic stress and wood anatomy of some lianas from Western Ghats Vidya Patil* and N.P. Malpathak Department of Botany, University of Pune, Pune-411007, India *Presenting author: [email protected]
Lianas constitute an important floristic component of tropical forests, (approximately 25%) where their high abundance has been used to differentiate tropical from temperate forests. Lianas are vines that begin life on the ground as small self- supporting shrubs and rely on other plants to reach the light-rich environment of the upper canopy competing directly with other trees for sunlight. They adapt themselves for support, climbing and twisting. As thickness increases in lianas, they show abnormal secondary growth and the storage organs exhibit anomalous structure. These anomalous structures may be directly influenced by external environmental conditions. Precise reason behind occurrence of cambial variants is yet not known, but it is considered that this is due to the effect of physiological or environmental stress. Acclimatized or adaptive structures like presence of wide vessels in xylem helps in high hydraulic conductivity. Abundance of parenchyma and ray parenchyma cells within and between xylem groups provides good flexibility with minimum damage to water conduits in the stem during climbing and twisted growth. It also limits physical disruption of vascular tissues during twisting and bending, and also promotes wound healing after girdling. It might also play a role in photosynthate storage. Present study focused on abiotic stress and their management and adaptations for growth and existence.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 146 Session A : Abiotic Stress Adaptation and Management
PA065 (IPPC0454) Analysis of genotypic variability for stem reserve mobilization in rice germplasm with contrasting drought tolerance A.S. Sakhare1*, Puja Rai1, Sudhir Kumar1, P.C. Nautyal2 and V. Chinnusamy1 1Division of Plant Physiology, 2Division of Seed Science and Technology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Non-structural carbohydrates accumulation in stem contributes substantially to grain development under stresses and hence, considered important for yield stabilization in grasses. In order to understand the contribution of non-structural carbohydrates remobilization in drought tolerance, we performed an experiment to analyze the genotypic variability for stem reserve mobilization (SRM) in rice germplasm with contrasting drought tolerance. Fourteen rice genotypes with contrasting drought tolerance were grown in the field and drought stress was imposed by withholding irrigation water. For estimation of SRM, plant samples were collected at flowering and at maturity stages. The relative change in dry weight and non-structural carbohydrates in stem, sheath, leaf and panicle tissues were analyzed to estimate SRM. Significant variability was observed among the rice genotypes for non-structural carbohydrates remobilization under drought stress. Under control conditions, reserve mobilization varied from 0 to 53%. Drought stress enhanced reserve mobilization in Crossa, MTU 1010, Vandana, CR-143-2-2 and Nagina 22 with maximum increase in CR-143-2-2. In case of Pusa 44, IR64 and Swarna, mobilization percentage decreased considerably under stress. Physiological and molecular basis of the genotypic differences in SRM are being investigated.
PA066 (IPPC0474) Characterization of a novel zinc finger transcription factor (TaZnF) conferring heat stress tolerance in plants Preeti Agarwal* and Paramjit Khurana Department of Plant Molecular Biology, University of Delhi, New Delhi-110021, India *Presenting author: [email protected]
High temperature is known to affect the crop production worldwide especially wheat. Under high temperature stress, a number of Transcription factors are activated including HSF’s, MYBs, bZIP in wheat. Recently, we have identified a zinc finger transcription factor in wheat from a suppression subtractive heat stress library. TaZnF belongs to C3CH4 type which constitutes the largest family in the plant kingdom and is reported to play important roles during various abiotic stress conditions. The characteristic feature of TaZnF includes a typical RING domain and a transmembrane region. The clustal alignment studies revealed high degree of conservation of zinc finger domain across various other orthologous members. TaZnF expression was upregulated under high temperature, salt, mannitol and cold stress. Transcriptional activation of TaZnF was confirmed by trans-activation in yeast. Localization studies revealed the possible localization to the nucleus and the membrane. The Arabidopsis transgenic plants over-expressing TaZnF showed high tolerance to high temperature stress at 42°C and performed better with respect to their growth and yield. Besides these responses to high temperature, the OE Arabidopsis transgenics showed considerable tolerance to drought and salinity stress. These observations suggest that TaZnF plays an important role during heat stress. Therefore TaZnF can be of great importance in improving thermal stress tolerance in plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 147 Session A : Abiotic Stress Adaptation and Management
PA067 (IPPC0480) Gene pyramiding of antioxidant enzymes leads to abiotic stress tolerance in Brassica juncea Neelam Prabha Negi1*, Vinay Sharma2 and Neera Bhalla Sarin1 1School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Department of Bioscience and Biotechnology, Banasthali University, P.O. Banasthali, Vidyapeeth, Rajasthan-304022, India *Presenting author: [email protected]
Reactive oxygen species (ROS) are generated by biotic and abiotic stresses such as drought, salinity, strong light, extreme temperatures, heavy metals, UV radiation, atmospheric contamination, mechanical wounding, nutrient starvation and pathogen attacks. Antioxidant enzymes play a significant role in eliminating toxic levels of reactive oxygen species (ROS), generated by living cells under stress. The antioxidative defense is initated by SOD, which converts superoxide - (O2 ) radicals to H2O2. The H2O2 that is also potentially harmful is converted to non-toxic water and monodehydroascorbate by the APX enzyme utilizing ascorbate as the electron donor. In the present study, genes for two different cytosolic antioxidant enzymes, superoxide dismutase (AhCuZnSOD) and ascorbate peroxidase (AhcAPX) isolated from salt tolerant cell lines of Arachis hypogaea were simultaneously over expressed in highly important oilseed crop, Brassica juncea (Indian Mustard) to check the effect on alleviation of abiotic stress. Compared with wild type plants, transgenic plants survived under longer period of water deficiency and salinity stress and displayed improved recovery after rehydration. The enhanced levels of antioxidant enzymes in the transgenic plants correlated with higher relative water content, improved photosynthetic efficiency, less electrolyte damage, elevated accumulation of compatible osmolytes, less malondialdehyde - as well as H2O2 accumulation and O2 accumulation under stress conditions compared to untransformed wild-type controls. Our analysis showed that an increase in the activity of antioxidant enzymes can significantly enhance salt tolerance in transgenic plants, suggesting an important role of combined over expression of two antioxidant enzymes.
PA068 (IPPC0483) High temperature tolerance in chickpea (Cicer arietinum L.): Possible involvement of zeaxanthin cycle pigments Pramod Kumar*, Sanjay Yadav and Madan Pal Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Chickpea being a cool season crop often experiences abnormally high temperature during reproductive phase. Terminal high temperature stress is one of the major constraints for its production. In nature, plants have developed several mechanisms to combat the high temperature stress. Our approach leads to a consideration of the photosynthetic pigments for high temperature tolerance. So, in order to analyze the heat tolerance in relation to photosynthetic pigments, an experiment was conducted using 4 contrasting genotypes namely BG 240 and JG 14 (relatively heat tolerant), SBD 377 (moderately tolerant) and ICC 1882 (relatively heat sensitive). High temperature was imposed by delaying the sowing date i.e. normal (18th November) and late sown (18th December). Under late sown condition, high temperature was experienced by crop from flowering stage to crop maturity (during this period maximum temperature ranged from 25oC to 39.9oC). Under late sown high temperature condition, in general, high temperature tolerant genotypes BG 240 and JG 14 maintained higher level of membrane stability index, RWC (%), osmolytes, dry matter partitioning, grain yield and heat tolerance index and had higher values of zeaxanthin, quantum yield of PS II (Fv/Fm ratio), non-photochemical quenching (NPQ), photosynthetic rate, level of photosynthetic pigments (chlorophylls and carotenoids) and lower level of violaxanthin, and lipid peroxidation as compared to high temperature sensitive one (ICC 1882). Furthermore, Fv/Fm ratio and NPQ showed positive association with high temperature tolerance which in turn indicated the involvement of zeaxanthin cycle pigments in high temperature tolerance of chickpea.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 148 Session A : Abiotic Stress Adaptation and Management
PA069 (IPPC0484) Plasticity in root-shoot development under water stress: exploring genotypic variation in wheat progenitor species, Aegilops tauschii and Triticum dicoccoides Yadhu Suneja1*, Anil Gupta1 and Navtej Singh Bains2 1Department of Biochemistry, 2Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana-141004, Punjab, India *Presenting author: [email protected]
A set of fifty seven Aegilops tauschii (DD genome), twenty six Triticum dicoccoides (AABB genome) accessions, wild wheat progenitors, along with tetraploid durum and hexaploid wheat cultivars were evaluated for their responsiveness to water stress. Observations across stress and non-stress environments allowed elucidation of genotypic variability in the level of induced responses for various traits. Though under PEG-induced water deficiency, average root length decreased from 3.82 cm to 3.66 cm for Ae. tauschii and from 5.40 cm to 5.26 cm for T. dicoccoides, several accessions demonstrated root elongation. The spectrum of variation was wider for increase than decrease in root length and biomass. Thirty three Ae. tauschii accessions and eighteen T. dicoccoides accessions showed an increase in root dry biomass under stress that ranged from 2% to 295%. Shoot parameters- length and biomass, by and large were suppressed by water deficit stress. Change in root length had a significant negative correlation with change in shoot length (r= -0.519). Change in root dry weight was significantly negatively associated with change in shoot length (r= -0.242) and change in shoot dry biomass (r= -0.369). Cross talk of root-shoot development with proline dynamics was also investigated. Aegilops tauschii accessions- 9803, 9814, 14109, 14191 and T. dicoccoides accessions- 7130, 7056 that showed maximum root enhancement and minimum shoot retardation under stress are worthy targets of molecular genetic analysis and subsequent marker-assisted mobilization of appropriate levels of inducibility for root and shoot development in the elite wheat backgrounds.
PA070 (IPPC0495) Deep rooted crops under center pivot irrigation: managing water stress Sangamesh Angadi*, Sultan Begna, Sukhbir Singh and Khulbhushan Grover Plant and Environmental Sciences, New Mexico State University, 2346 SR 288, Clovis 88101, United States *Presenting author: [email protected]
Agriculture in the Southern Great Plains of the United States perennially suffers from abiotic stresses including water, wind and temperature. Irrigation water from the Ogallala Aquifer converted this “Dust Bowl of the 1930s” into a highly productive agriculture region in the country. Center Pivot Irrigation Systems was invented in the region to improve irrigation application efficiency and reduce energy costs, which is used extensively. However, continued decline of well outputs and need for crop diversification have led to assess deep rooted alternative crops for the region. Center pivot system simulates frequent rainfall events and applies small quantities of water frequently to wet a shallow depth of the soil profile. With the current practice of irrigation deeper soil profiles are less frequently used. A multiyear study using the deep rooted safflower crop was conducted to assess effects of storing water in the deep soil profile using pre-season irrigation, varying in-season irrigation levels and critical growth stage based irrigation management on safflower stress physiology, seed yield formation and productivity. Safflower responded to water stress with multiple strategies. It ranged from morphological adjustment of plant size and leaf area to physiological response of stomatal and photosynthesis regulation. Safflower relied on its strong root system to extract moisture from depth. Although the amount of water extracted from depth was relatively small, the efficiency of using that water in yield formation was much higher.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 149 Session A : Abiotic Stress Adaptation and Management
PA071 (IPPC0496) Interplay between aldo-keto reductase (AKR) and glyoxalases as an important strategy for methylglyoxal detoxification during abiotic stress in Jatropha curcas L. Shalini Mudalkar* and Attipalli Ramachandra Reddy Department of Plant Sciences, University of Hyderabad, Hyderabad-500046, India *Presenting author: [email protected]
Methylglyoxal (MG), a by-product of oxidative stress, is responsible for protein oxidation, DNA damage and lipid peroxidation in plants leading to the generation of advanced glycation end products (AGEs). The accumulation of MG was studied in Jatropha curcas under progressive salinity and drought stresses, wherein the levels of MG increased with increasing stress severity. To understand the mechanism of MG detoxification and plant protection in J. curcas under abiotic stress, aldo-keto reductase (JcAKR) and glyoxalases (JcGLY1 and JcGLY2) were cloned, characterised and heterologously expressed in E. coli BL-21 (DE3). MALDI-TOF analysis was performed to confirm the purified proteins, which showed significantly high enzyme activities and catalytic efficiencies with methylglyoxal (MG) as substrate. The expression of JcAKR, JcGLY1 and JcGLY2 determined through qRT-PCR analysis, showed up-regulation in leaf and root tissues under progressive drought and salinity stresses. To further confirm the protective roles of JcAKR, JcGLY1 and JcGLY2, spotting assays with supplemental PEG (5%), NaCl (200 mM) and MG (5 mM) were performed with E. coli and yeast. The bacteria and yeast harbouring the respective genes were able to grow in the presence of external stress compared to controls. In conclusion, JcAKR, JcGLY1 and JcGLY2 act together to protect the plant from the harmful effects of MG under conditions of abiotic stress.
PA072 (IPPC0498) Genetic engineering of elite rice cultivars by inducible expression of late embryogenesis abundant gene(s) from wild genotype, O. nivara improves tolerance towards soil water stress without yield penalty Ranjeet Kaur*, Soumitra Kumar Sen and Ananta Kumar Ghosh Indian Institute of Technology, Kharagpur-721302, West Bengal, India *Presenting author: [email protected]
Soil water stress (SWS) is one of the major environmental stress conditions that limits crop yield. Inducible expression of gene(s) conferring tolerance to SWS is desirable for producing farmer-friendly stress tolerant crop plants. Gene(s) encoding late embryogenesis abundant (LEA) proteins could be a prospective candidate in this regard. A comparative expression profiling of Wsi18, a group 3 LEA gene, performed by means of stage specific genome wide microarray and subsequent real time qPCR analysis, revealed differential response under SWS conditions across different rice genotypes. Genetic polymorphism in the coding DNA sequence as well as in the 5' regulatory elements of Wsi18 were looked into between wild rice and elite cultivars and it revealed interesting insight into their evolutionary structure-function relationship. Wsi18 promoter from Oryza nivara (wild progenitor of cultivated rice) showed enhanced expression of the reporter gusA gene in comparison to its orthologue from IR20. A further attempt was undertaken to functionally ascertain the role of such variant allelomorphic forms to influence the capacity to withstand SWS condition of any stress sensitive elite rice cultivar in terms of their ultimate grain yield via transgenic approach. When driven by the endogenous OnWsi18 promoter, the transgenic IR20 rice plants carrying the OnWsi18 gene showed improved tolerance towards SWS without any yield penalty. The study, to the best of our belief, is expected to contribute to the development of molecular strategies to generate SWS tolerant rice lines that can produce well.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 150 Session A : Abiotic Stress Adaptation and Management
PA073 (IPPC0511) Drought stress impact on physiological, biomass and yield parameters- genotypic variability in maize P. Vagheera, R. Abdul, M. Vanaja*, M. Maheswari, P. Sathish, N. Jyothi Lakshmi, G. Vijaya Kumar, S.K. Yadav and B. Sarkar Central Research Institute for Dryland Agriculture (CRIDA), Santoshnagar, Hyderabad, India *Presenting author: [email protected]
Five maize (Zea mays L.) genotypes- DTL-3, DTL-4, DTL-4-1, DTL-9 and DTL-11 from CRIDA maize germplasm were evaluated along with drought tolerant genotype Varun for their response to drought stress at flowering and to identify the traits contributing to it. The moisture deficit stress was imposed at initiation of anthesis and stress was relieved once wilting symptoms appeared. The measurements of photosynthesis and transpiration were recorded at stress point and after stress release and compared with well watered controls. Genotypes DTL-3 and DTL-4-1 recorded higher photosynthetic rate, lower stomatal conductance, lower transpiration rate and higher WUE even under drought stress and stress released conditions. With imposition of drought stress, photosynthetic rate reduced from 38% (DTL-3) to 62% (Varun), however DTL-3 and DTL-11 recovered to normal when stress was relieved. Drought stress decreased total biomass of maize genotypes from 6% (DTL-4-1) to 29% (DTL-11) due to decreased stem and cob biomass. Among the genotypes, DTL-4 recorded highest per se total biomass and seed yield under both irrigated and stress condition. Drought stress reduced seed yield (7% to 43%) and the reduction was mainly due to decreased seed number (6% to 44%) while change in test weight was non-significant. DTL-4 performed best among the selected genotypes both under well watered and drought stress condition due to its better ability to partitioning of biomass to shoot and reproductive parts.
PA074 (IPPC0512) Hydraulic dynamics were associated with expression of major intrinsic proteins (MIPs) in mulberry (Morusspp L.) subjected to progressive drought stress Kanubothula Sita Rami Reddy* and Attipalli R. Reddy Department of Plant Sciences, University of Hyderabad, Hyderabad-500046, India *Presenting author: [email protected]
Two mulberry genotypes including a drought tolerant (DT), Selection-13 (S13), and a drought susceptible (DS), Kanva-2 (K2), were subjected to progressive drought stress to elucidate the hydraulic dynamics in relation to changes inphotosynthetic leaf gas exchange characteristics as well as to understand the expression patterns of major aquaporins. Drought imposition induced severe reduction in photosynthetic performance in both S13 and K2 genotypes after 15 days of stress (DAS) than their respective controls. However, S13 showed better photosynthetic rate (Pn) and water use efficiency (WUEi), with minimal reduction in stomatal conductance (gs) and transpiration (E). These characterizations of photosynthesis and water use efficiency in turn were associated with greater leaf water potential (øL), sap flow (F), leaf
(KL) and stem (KS) hydraulic conductance compared toK2 at 15 DAS. Further, S13 showed an upregulation of mRNA transcripts of MIP isoforms including plasma membrane intrinsic proteins (PIPs) and tonoplast intrinsic proteins (TIPs) in both leaf as well as in roots after 15 DAS. In conclusion, plant hydraulics and the expression levels of certain MIPs should serve as desirable traits in crop improvement programs for drought tolerance.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 151 Session A : Abiotic Stress Adaptation and Management
PA075 (IPPC0518) Proteogenomic landscape of an orphan legume, grasspea, challenged with dehydration stress Divya Rathi*, Subhra Chakraborty and Niranjan Chakraborty Plant Stress Laboratory, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Plants respond and adapt to adverse environmental stresses by altering their cellular metabolism and activating various defense machineries. Investigation of genetic resource for stress adaptation, in plants, is crucial not only for better understanding the underlying mechanism, but also as an attractive target for improving stress tolerance. Grasspea, often regarded as an ‘orphan’ legume, serves as a model system for rich source of nitrogen beneficial for soil fertility, and is a valuable source of dietary protein. Further, many of its cultivars are capable to withstand a myriad of constraints, making it a suitable model species to study stress tolerance. As part of proteomics studies on legumes, we reported earlier the salinity- and cold-responsive proteome of grasspea. In the present study, 3-week-old grasspea seedlings were subjected to dehydration for 144 h. There were no visible changes in the seedlings till 48 h of dehydration. The leaflets rolled after 60 h and displayed chlorosis, and the damage aggravated further during 72-144 h. The physiological changes of stressed seedlings were thereby monitored. The temporal effects of dehydration on grasspea were evaluated at the proteomics as well as transcriptomics levels using classical 2-DE and RNA-seq, respectively. The 2-DE analysis revealed approximately 200 proteins that were dehydration-responsive and were differentially regulated in a qualitative as well as quantitative manner. Conversely, approximately ten-fold numbers of transcripts were found to be dehydration-responsive. This comparative stress-responsive proteome of grasspea would provide new information that would be used for targeted breeding approaches for crop improvement.
PA076 (IPPC0522) Specific leaf weight: Better indicator to screen drought tolerance in cotton genotypes S.B. Amarshettiwar*, T.H. Rathod, G.R. Shamkuwar and W.K. Shembekar Department of Agricultural Botany, PGI, Dr. PDKV, Akola-444104, India *Presenting author: [email protected]
Present investigation was conducted during kharif season of 2010 and 2011 in RBD on the experimental field under non stress and water stress conditions in pot culture. The seeds of 31 cotton genotypes (10 G. arboreum and 21 G. hirsutum) were sown in three replications. Experimental findings are based on two years pooled data under water stress and non stress conditions. Results indicate significant superiority of G. arboreum over G. hirsutum, which indicates its tolerance towards water stress on the basis of morpho-physiological and seed cotton yield. Result revealed that Under water stress condition, five G. arboreum genotypes, viz., AKA-8 (65.03 g/plant), AKDH-1 (62.30 g/plant), AKDH-5 (55-69 g/plant), Garrohill (55.15 g/plant), GAK-423 (54.97 g/plant) and three G. hirsutum genotypes, viz., PKV Hy-2 (58.22 g/plant), PKV- Hy-5 (57.87 g/plant), PKV-Hy-4 (52.43 g/plant) recorded significantly higher seed cotton yield, which were 46.7, 40.54, 25.63, 24.41, 24.0, 31.33, 30.54 and 18.27% higher than general mean (44.33 g/plant) of both the species, respectively. At 120 DAS, both under stress and non stress conditions, G. arboreum genotypes showed significantly higher SLW than G. hirsutum. Specific leaf weight was found to be best indicator for drought tolerance. Among G. arboretum, AKA-8 (1.78 g/dm²) Garrohill (1.91 g/dm²), AKDH-1 (1.82 g/dm²), KWA-8 (1.53 g/dm²), HD-110-151 (1.50 g/dm²), AKA-7 (1.49 g/dm²) and among G. hirsutum, PKV-Hy-4 (1.51 g/dm²), PKV Hy-5 (1.34 g/dm²), Yamuna (1.17 g/dm²) and PKV-Hy-2 (1.10 g/ dm²) were found more tolerant genotypes for water stress condition. The correlation co-efficient revealed positive correlations between seed cotton yield and SLW (r=0.741),+ and this criteria may be useful for assessing genotypes for drought studies.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 152 Session A : Abiotic Stress Adaptation and Management
PA077 (IPPC0525) Screening of cotton (G. hirsutum L.) germplasm for adaptation to drought stress resilient Jayant Meshram* and J. Annie Sheeba Crop Production Division, ICAR-Central Institute for Cotton Research, Nagpur-440010, India *Presenting author: [email protected]
Though cotton is drought tolerant crop, drought is a major problem in rain-fed areas. . Leaf reddening and wilting are also getting more pronounced in recent years, following drought stress. A field experiment was conducted under rain-fed and irrigated condition at ICAR-CICR, Nagpur during 2014-15 to study drought tolerance traits in twenty cotton genotypes on the basis of various stress indices for yield and yield attributing characters. The physiological changes induced due to water stress were quantified by withholding irrigation for continuous 25 days to 45 days old cotton plants in July sown crop. In all twenty genotypes comprising Bt cotton (Mallika), non-Bt cotton (var. Suraj), susceptible and drought tolerant genotypes were evaluated for leaf area, leaf relative water content, mid-day water potential, epicuticular wax in the leaves, plant height, number of bolls per plant, boll weight (g) and correlated with temperature and volumetric soil moisture content. Data revealed that Nagpur 9, F-1226, and SGNR-27 genotypes recorded highest epicuticular wax content and yield attributing characters under both irrigated and rain-fed conditions. While IC-357045 germplasm accession showed reduction in drought tolerance traits and seem to be susceptible to moisture stress. A field phenotyping approach is to initially identify promising germplasms with high cellular tolerance to drought stress to withstand dry spell for improving field level drought tolerance. The differences of root traits in terms of drought tolerance will enable to augment cotton productivity under water limited environments but combining these traits with high yield potential is required.
PA078 (IPPC0527) A Class-I TCP protein modulates ABI4-mediated pathways and lipid droplet synthesis to influence developmental and abiotic stress signaling Pradipto Mukhopadhyay* and Akhilesh Kumar Tyagi National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Abiotic stresses severely affect development of plants and facilitate the production of stress-related factors at the cost of growth-related proteins. A balance between these two classes of proteins determines the stress tolerance limit of the plants. TCP transcription factors form an indispensible group of plant-specific developmental regulators. We identified a stress-upregulated Class-I TCP transcription factor in rice, OsTCP19, the expression of which was found to be higher in stress-tolerant varieties under dehydration and high salinity. This gene also presents a case of tissue, variety and stress dependent alternative splicing. Overexpression of this gene not only affects the expression auxin, abscisic acid and jasmonate-dependent pathways but also increases the synthesis of lipid droplets and related genes in Arabidopsis. The transgenic Arabidopsis plants show many developmental alterations in addition to increased tolerance to abiotic stress. In silico analysis also predicts OsTCP19-mediated similar alteration in gene expression in rice. This protein was further found to interact with OsABI4 and OsULT1, a transcriptional co-regulator, and directly activate a triacyglycerol biosynthesis gene in transient assays. Together, these results suggest OsTCP19 as an important regulator which crosslinks developmental and abiotic stress signaling.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 153 Session A : Abiotic Stress Adaptation and Management
PA079 (IPPC0536) Transcriptome analysis of salt susceptible blackgram (Vigna mungo L. Hepper) under saline conditions Pooja Bangar1,2*, Ashok Chaudhury1 and K.V. Bhat2 1Bio & Nano Technology, Guru Jambheshwar University of Science and Technology, Gurgaon-122002, India 2National Bureau of Plant Genomic Resources, Pusa Campus, New Delhi-110012, India *Presenting author: [email protected]
Among various abiotic stresses, salinity is one of the world’s most serious problems as plant agriculture faces a loss of $12 million annually due to salinity stress worldwide (FAO stat 2006). Blackgram (Vigna mungo L. Hepper, 2n=22) an important grain legume grown for its protein rich edible dry seeds in the tropical and subtropical regions of the Indian subcontinent, is adversely affected by salinity stress. We aimed to identify differentially expressed genes (DEGs) and its transcription factors by comprehensive analysis of transcriptomes of both root and leaf tissues of Blackgram genotype grown under the control and saline conditions. A total of427 million high quality reads were generatedby employing Illuminamiseq sequencing platform which were assembled (de nova) using CLC Genomics Workbench 7.5.1 at default parameters into 591,822 contigs with an average length of 300 bp and N50 value of 369 bp. Functional classification revealed 65.83% unique contigs into 20 subcategories of the biological process, 32.77% unique contigs into 14 subcategories of molecular functions and 37.80% unique contigs into 11 subcategories of cellular components. We identified 5659 candidates salt responsive genes in which 1259 are upregulated and 4400 are down regulated genes (with a RPKM fold change >2 and a cut off of p <0.001) with significantly different expression under salt water and fresh water treatments. The data generated in this study will help in understanding the fundamentals of salt tolerance in blackgram and will provide resources for functional genomics studies and in crop improvement.
PA080 (IPPC0559) Oxidative stress tolerance correlates with high temperature stress tolerance at plant level in rice P. Boominathan*, R. Sivakumar and V. Ravichandran Department of Crop Physiology, Tamil Nadu Agricultural University, Coimbatore–641003, Tamil Nadu, India *Presenting author: [email protected]
Among the many challenges to achieve higher productivity in rice, climate change associated phenomenon especially high temperature is increasingly becoming an important factor. Most of rice is currently cultivated in the regions where temperature is close to optimum or above for growth. Therefore, any further increase in mean temperature would certainly reduce the rice yield. One of the most common high temperature stress effects at cellular level is the oxidative stress caused by various reactive oxygen species. An investigation was conducted with eleven genotypes including popular varieties of Tamil Nadu, landraces and tolerant and susceptible cultivars known for the high temperature tolerance with the hypothesis that oxidative stress tolerant genotypes would also tolerate abiotic stresses including high temperature stress. To assess the genetic variability in response to oxidative stress, rice seedlings were treated with a quinine, known as menadione, to generate reactive oxygen species. The duration of exposure and concentration of menadione optimized as 2 mM for 24 hrs in rice. The study revealed that the genotypes white Ponni, Moroberekan, Norungan which showed higher sensitivity to high temperature in the growth chamber at plant level, also exhibited higher percent reduction in survival during menadione treatment. Thus, developing the oxidative stress tolerant genotypes would eventually helps in identifying genotypes tolerant to high temperature stress. The tolerant genotypes displayed lower membrane injury thus strongly supporting the fact that membranes are primary sites for oxidative stress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 154 Session A : Abiotic Stress Adaptation and Management
PA081 (IPPC0568) Influence of waterlogging on physio-biochemical traits and expression behavior of SOD and ADH genes in sugarcane genotypes Radha Jain1*, S.P. Singh1, Amaresh Chandra1, Anshu Singh1, Smita Singh1, R.K. Singh1, A.D. Pathak1, M. Swapna1 and V.K. Srivastava2 1Plant Physiology and Biochemisrty Division, Indian Institute of Sugarcane Research, Lucknow-226002, U.P., Indian Uttar Pradesh Council of Sugarcane Research, Shahjahanpur, U.P., India *Presenting author: [email protected] Waterlogging is one of the serious abiotic stresses affecting sugarcane productivity. It usually occurs due to rise in water table in almost the entire canal irrigated areas during rainy season (July-September) in North Bihar, Eastern Uttar Pradesh, West Bengal, coastal areas of Orissa, Andhra Pradesh and Kolhapur area of Maharashtra. Cane yield and juice quality losses due to waterlogging depend on genotypes, environmental conditions, development stage and inundation duration. In present study, an effort was made to characterize sugarcane genotypes for waterlogging tolerance based on physio- biochemical traits. Three bud setts of twenty four sugarcane genotypes including six commercial cultivars, CoLk 94184, BO 91, CoS 767, CoJ 64, CoS 97264 and UP 9530 were planted at Kharika Block, Indian Institute of Sugarcane Research, Lucknow during spring season under two different conditions; control (non-waterlogged) and waterlogged. In waterlogging treatment, crop was grown in deep plot which was naturally waterlogged during rainy season. Results obtained indicated decrease in leaf area, SPAD reading, chlorophyll a, b, carotenoids, specific leaf weight (SLW), nitrate reductase (NR) activity under waterlogged conditions. Almost all the genotypes exhibited aerial rooting due to waterlogging. Genotypes varied significantly for SLW, SLA, SPAD and NR activity. Quality attributes; oBrix, sucrose and juice purity were decreased due to waterlogging; only a few genotypes showed slight increase over control. Waterlogged affected plants exhibited higher expression of ADH and SOD genes in leaf, root and stalk; ADH expression was significantly high in roots. Present findings help in identifying and characterizing sugarcane genotypes tolerant to waterlogging.
PA082 (IPPC0585) OsHsfA6a directly regulates a subset of Hsp genes and functions as omphalos for transcriptional regulation of ClpB/Hsp100 genes in rice (Oryza sativa L.) Dhruv Lavania*, Anirban Chakraborty, Anuradha Dhingra, Juarez Road, Dhaula and Anil Grover Department of Plant Molecular Biology, University of Delhi, South Campus, Benito Juarez Road, Dhaula, New Delhi-110021, India *Presenting author: [email protected]
ClpB/ Hsp100 proteins are critically important in governing heat stress tolerance in plants. The expression of OsClpB-C (cytoplasmic isoform) is mainly regulated at the transcriptional level. We have earlier shown two important events in this process: (a) OsHsfA6a regulates OsClpB-C promoter and (b) OsHsfA6a interacts with OsClpB-C and OsHsfB4b proteins. In this study, OsHsfA6a was identified as the only direct transcriptional activator of OsClpB-C and OsClpB-P (plastidial isoform) genes and as direct transcriptional activator of a subset of heat inducible OsHsp genes. The transcriptional activity of OsHsfA6a was repressed in reporter gene assays by interactions with OsClpB-C and OsHsfB4b proteins. Protein-protein interaction analysis revealed heteromeric interactions of OsHsfA6a with several class A OsHsfs and the complexes were nuclear localized. This is the first study on heteromeric interactions of subclass A6 Hsf member and the highest number of heteromeric interactions for any plant Hsf to date. The interacting OsHsfs showed differential transcript expression kinetics under a time-course regime of heat stress and recovery. We thus infer that the target gene selectivity and activity of OsHsfA6a may be regulated by virtue of these interactions at post-translational level. Our findings suggest that binding of OsClpB-C with OsHsfA6a may work in a negative feedback-loop manner and OsHsfB4b might function as a repressor protein. We conclude that a supra-complex involving several OsHsfs regulates Hsp promoters under heat stress and recovery in rice.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 155 Session A : Abiotic Stress Adaptation and Management
PA083 (IPPC0598) Better antioxidative defense mechanism of rainfed lowland rice varieties is associated with their tolerant to submergence Ashish Kumar Srivastava1*, Akhtar Hussain Khan2, Phool Chand Ram2, Sudhanshu Singh1 and Abdelbagi M. Ismail3 1Plant Breeding Genetics and Biotechnology, International Rice Research Institute, New Delhi-110008, India 2Narendra Deva University of Agriculture and Technology, Kumarganj, Faizabad, UP, India 3International Rice Research Institute, Los Baños, Philippines *Presenting author: [email protected]
21-days-old seedlings of 4 tolerant (FR13A, Swarna-Sub1, NDR9730018, NDR9930111) and 2 susceptible (Swarna and IR42) lowland rice varieties were submerged for 10 days in outdoor flooded water tank. Same set of lines were planted without imposing submergence to record grain yield under natural conditions. Enzymes of Halliwell-Asada pathway (SOD, APX, GR) were estimated before submergence and 0, 2, 4, 6 and 24th hours after de-submergence, whereas enzyme CAT and antioxidants (AsA and GSH) were estimated before and just after de-submergence. SOD activity increased (3- 4 fold in tolerant vs 1-1.5 fold in susceptible) up to 4th hr after de-submergence and subsequently declined till 24th hr. Upon de-submergence successive increase in APX activity was recorded up to 24th hr in all varieties. Tolerant genotypes showed consistently higher GR activity after de-submergence. Higher CAT activity was recorded in tolerant varieties after submergence (48.8-58.2%) when compared to IR42 and Swarna (28.4 and 38.3%). AsA and GSH contents showed opposite trend after de-submergence, IR42 and Swarna showing 7.6 and 14.7% increase in AsA values and more than 80% reduction in GSH values. After submergence, FR13A and Swarna-Sub1 recorded more than 85% survival, whereas IR42 and Swarna showed only 41.5% and 61.5% survival. FR13A and Swarna-Sub1 exhibited only 41.7 and 50% reduction in grain yield, whereas Swarna and IR42 showed 60 and 68% reductions when compared to their yield in natural conditions. It can be thus concluded that better antioxidative defense mechanism of tolerant lines is associated with better survival and higher grain yield under submergence.
PA084 (IPPC0617) Identifying plant genetic resources adapted to elevated temperature Sunil Archak1*, Anuj Singh1, Sarika Mittra2, D.P. Semwal1, P.N. Mathur3, P.K. Aggarwal4 and K.C. Bansal1 1ICAR-National Bureau of Plant Genetic Resources, New Delhi-110012, India 2Bioversity International, New Delhi-110012, India 3Regional Representative, Central and South Asia, Bioversity International, New Delhi-110012, India 4Regional Program Leader South Asia, CCAFS, New Delhi-110012, India *Presenting author: [email protected]
Climate smart agriculture hinges on the cultivars with greater adaptability and resilience. This in turn necessitates the use of a wider range of intra-specific diversity conserved in the genebanks. Estimation of adaptive capacity of genebank accessions based on passport data and identification of germplasm accessions with excellent adaptation potential can help develop climate smart crop varieties. Globally, genebanks are working to bring together the inherent diversity of ex situ collections and power of climate analogues for enhancing adaptive capacity of food crops to climate change. We attempted to employ climate tools to identify pre-adapted germplasm among ~39,000 genebank accessions belonging to five select crops- wheat, pearl millet, chickpea, pigeon pea and sorghum. The methodology comprised geo-referencing and clustering the accessions, climate matching, designating pre-adapted material and collecting germplasm from predicted sites.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 156 Session A : Abiotic Stress Adaptation and Management
PA085 (IPPC0621) Overexpression of AtICE1 gene improves abiotic stress tolerance in transgenic rice Rakesh Verma1*, Santosh Vinjamuri1, Shashank Yadav1, M.V. Rao2 and Viswanathan Chinnusamy1 1Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India 2Department of Plant Sciences Bharthidasan University, Tiruchirappalli, Tamil Nadu, India *Presenting author: [email protected]
Abiotic stresses such as drought, soil salinity and freezing, greatly affect plant productivity and quality. Many genes are involved in the process of adaptation to adverse environmental conditions. ICE1 (Inducer of CBF Expression 1), an upstream transcription factor, regulates the transcription of CBFs and other genes, and confers freezing tolerance to Arabidopsis. Recent studies from different plant species suggest that ICE1 also regulates tolerance to other abiotic stresses. Arabidopsis ICE1 gene was isolated by RT-PCR and cloned in a modified pCAMBIA1300 under stress inducible AtRD29A promoter and transformed into indica rice variety Pusa Sugandh 2 by the Agrobacterium-mediated transformation. Cold, drought and salt stress tolerance of transgenic events were compared with the non-transgenic plants in seedling, vegetative and flowering stages. Transgenic plants exhibited enhanced tolerance to these stresses in terms of RWC, chlorophyll and membrane stability. In addition, transgenic rice plants overexpressing AtICE1 exhibited higher stomatal density but smaller stomatal size as compared to non-transgenic plants. Further, leaf width and length were significantly lower in transgenic plants. The results suggest that the function of AtICE1 in stress tolerance and stomatal development is conserved in rice and the over-expression of ICE1 can improve tolerance to abiotic stresses in rice.
PA086 (IPPC0625) Adverse effects of short exposure to high temperature on wheat and mustard and its amelioration through iron and zinc application Uday Burman* and Praveen Kumar Division of Integrated Land Use Management & Farming System, Central Arid Zone Research Institute, Jodhpur-342003, India *Presenting author: [email protected]
Changes in temperature significantly influence plant processes and subsequent crop productivity. Temperature increased by 3-5 OC when plants are partially covered with <100 μm thick transparent polythene sheet. This resulted in 69.0, 70.7, 45.8 and 59.5 % reduction in plant water potential, nitrate reductase (NR) activity, total soluble carbohydrate (TSC) and starch in mustard. Corresponding values in wheat were 40.0, 39.3, 5.5 and 13.1%, respectively. In a two year study, seeds of both mustard (var. Pusa Jai Kisan) and wheat (var. Raj 3077) were primed with FeSO4 (10%), Fe EDDHA (0.6%),
ZnSO4 (36%) and Zn EDTA (6.0%) prior to sowing in pots. In another experiment, 0.125, 0.250, 0.50 and 1.00 % iron and zinc sulphate were foliar sprayed at pre flowering stage in field. Thereafter plants were subjected to high temperatures at flowering stage for seven days in both the experiments. Priming with 0.6% Fe EDDHA was associated with highest increase in RWC (30.7%), TSC (26.5%) and starch (68.0%) compared to corresponding control in wheat. Similar response was observed in mustard also. Foliar spray of 0.125% Zn was associated with less reduction in yield upon exposure to high temperature compared to control in both wheat and mustard. High NR activity (21% more than control) in wheat and maintenance of high membrane stability index in mustard could be responsible for the partial alleviation in yields.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 157 Session A : Abiotic Stress Adaptation and Management
PA087 (IPPC0627) Phenotypic traits as selection tools for screening mango rootstock against salt stress V.K. Singh* and S. Rajan Crop Production Division, ICAR-Central Institute for Subtropical Horticulture, Lucknow-226101, India *Presenting author: [email protected]
Mango is highly sensitive to soil salinity and alkalinity and tolerant rootstocks have potential for making mango cultivation possible in these problem soils. Keeping this in view, poly-embryonic cultivars as rootstock, viz., Bappakai, Goa, Kurukkan, Moovandon, Mylepelian, Nekkare, 13-1, Kitchner, Peach, Starch, and Chandrakaran, Olour, Phillippino and Vellaikulamban were evaluated under salt stress conditions. Symptoms of salt injury in seedlings appeared within one month and at higher EC (2-4 dS m-1), all plants died except Nekkare, 13-1 and Kurukkan. Salinity factor index revealed that 13-1 has better tolerance to salt stress and can survive at high level of salinity. With increasing salinity levels, highest reduction in root/shoot ratio was recorded in Goa and lowest in 13-1. Electrolyte leakage showed severe membrane damage in the stressed plants with drastic reduction in chlorophyll fluorescence and PSII photochemistry in majority of the rootstocks. Proline and glycine-betaine content were higher in leaves (4.5–6.0 mg g-1 dry wt.) than roots (1.2–3.5 mg g-1 dry wt.) with elevated salt stress, and results indicated low contribution towards osmotic adjustment. Leaf water potential was drastically ψ reduced with greater reduction (-19.78 w) in Starch and Kitchner, whereas, Kurukkan, 13-1 and Nekkare exhibited least reduction. Rootstock 13-1 exhibited higher cuticular wax (608.50 μg cm-2) content even in control (normal soil) plants, however, Kurukkan and Nekkare exhibited higher level of leaf wax when they were exposed to salt stress. Thus, these traits may be utilized for screening polyembryonic mango cultivars for rootstock purpose against salt stress.
PA088 (IPPC0630)
Effect of salinity on morphological traits of F2 population in rice P. Pundir*, S.L. Krishnamurthy, P.C. Sharma and D.K. Sharma Department of Biotechnology, Central Soil Salinity Research Institute, Karnal-132001, India *Presenting author: [email protected]
Salinity causes significant yield reduction in rice. There are substantial opportunities to improve grain yield by using conventional breeding under stress conditions. The F2 segregating population (PS 5/CSR10) of rice was studied under saline stress and non stress conditions at reproductive stage. The objective of this study was to unveil the performance -1 of same F2 progenies under both saline and non stress environment (EC ~ 10 dSm ) and to find out the suitable stress indices. Significant variability was found in segregating population for grain yield and other traits under both saline and non-saline condition. There was a mean reduction of 89% in grain yield due to high salt stress (EC~10 dsm-1). Under non- stress environment the significant positive correlation was noticed between grain yield and biomass (r=0.64) followed by grain yield with plant height (r=0.36) and grain yield with productive tillers (r=0.30). Under salinity weak and insignificant -1 association of grain yield was observed with other traits. Grain yield of the F2 population ranged from 400 to 9333 kg h under normal environment while it reduced to 40-1591 kg ha-1 under salinity. Grain yield and biomass were the affected traits most by salinity. Skewness and Kurtosis indicates the presence of complementary gene interaction under both environments thus population can be further improved for selection of salt tolerant lines. Correlation coefficient and biplot analysis of stress indices indicated that GMP, STI, MP.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 158 Session A : Abiotic Stress Adaptation and Management
PA089 (IPPC0641) Elucidating dominant enzyme mediated oxidative stress tolerance pathway in groundnut under salinity stress Koushik Chakraborty*, Sujit Bishi, Kiran Patel, Nisha Goswami, Debarati Bhaduri, A.L. Singh, Pratap V. Zala Directorate of Groundnut Research, Junagadh Agricultural University, Junagadh-362001, Gujarat *Presenting author: [email protected]
Salinity, a major abiotic factor limits plant growth and productivity globally. Oxidative stress, aroused due to higher accumulation of reactive oxygen species (ROS), restricts plant metabolism under salt-stress. Previous reports suggested existence of more than one mechanism for ROS-detoxification adapted by plants under abiotic stress. Hence, the present study focuses on elucidating major oxidative stress tolerance pathway in groundnut under salinity stress using differentially salt-sensitive genotypes. For this, a pot experiment was conducted with 6 genotypes (‘CS 240’, ‘NRCG 357’, ‘TMV 2’, ‘Girnar 1’, ‘TPG 41’, ‘Somnath’) and four different levels of salt treatment (T1:0, T2:25, T3:50 and T4:100 mM NaCl). Imposition of salinity treatments resulted in significant stress build-up evident from higher soil ECe value of 3.7 and 5.4 dS m-1 in T3 and T4, respectively and significant drop in leaf water potential (-15.5 MPa in control to -40 MPa under T4). Severe plant mortality was observed at T4 treatment. Determination of ROS level build-up by both spectrophotometric and dye-staining method revealed comparatively higher oxidative stress mediated damage in ‘NRCG 357’ and ‘TMV 2’ than ‘Somnath’ and ‘CS 240’. Activity assay of major ROS-detoxifying enzymes (superoxide dismutase, catalase, peroxidase, glutathione reductase, ascorbate peroxidase) and their relative transcript abundance through real-time qPCR technique revealed up-regulation of SOD, CAT and POD under salt stress, while no significant induction observed in GR and APX. Thus present study concludes, minimal role of ascorbate-glutathione mediated ROS-detoxification pathway and major dependence on SOD, CAT, POD pathway for oxidative stress tolerance in groundnut under salinity stress.
PA090 (IPPC0646) Phenotyping of rice genotype for high temperature stress tolerance at anthesis stage Sourabh Karwa*, Ashish K. Chaturvedi, Divya Shah, Shashi Meena and Madan Pal Singh Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Rising temperature is one of the most important climate factors affecting rice yield .Rice is considered sensitive to high temperature stress at flowering stage. High temperature can affect rice grain quality. This study reports phenotyping of thirty-six rice genotype raised in pots under ambient environment and exposed to high temperature stress (>38-40p C) after anthesis till maturity of crop. After ten days exposure to high temperature, spikelets were collected from control and high temperature stressed plants and pollen viability and fertility were analyzed using fluorescent microscopy. Thermal images of the plants were captured using IR thermal image camera and CTD was calculated. Spikelet fertility and panicle yield was calculated after harvest of crop. There was large variability in spikelet fertility among the genotypes, and it was significantly correlated with grain yield per plant. Based on the spikelet fertility, ten contrasting genotypes were identified. Tolerant rice genotypes showed higher pollen viability and fertility (65-80%) under high temperature, while in sensitive genotypes both the traits showed up to 70% reduction. Similarly, tolerant genotypes showed 3.4-5.5p C CTD under high temperature stress. All the genotypes will be characterized on physiological basis of high temperature tolerance and most tolerant genotypes will identified for future breeding program of rice crop.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 159 Session A : Abiotic Stress Adaptation and Management
PA091 (IPPC0667) Evaluation of walnut (Juglans regia) genetic diversity in central Himalayan region for moisture stress and quality A.K. Trivedi1,2*, S.K. Verma1 and R.R. Arya1 1ICAR–National Bureau of Plant Genetic Resources, Regional Station Bhowali District Nainital-263132, Uttarakhand, India 2Present Address: ICAR–Central Institute for Subtropical Horticulture, Rehmankhera, Lucknow-226101, Uttar Pradesh, India *Presenting author: [email protected]
Walnut (Juglans regia L.) is an important temperate nut grown in India. Walnut cultivation is sensitive to abiotic stresses, particularly moisture stress at seedling stage. Dry weather reduces potential productivity of walnuts even in the absence of soil water deficit. Although, it is one of the major nut fruit grown in Himalayan region but it has remained a low priority crop in temperate fruit growing areas of India. Genetic diversity of walnuts available in the Himalayan region, which may possess valuable stress resistance that helps them to cope with the unfavourable environmental conditions. Hence, in order to collect available genetic diversity of walnut, germplasm survey and exploration expeditions were conducted in central Himalayan region. A total of 15 accessions having distinct traits of horticultural importance were collected and conserved in the field gene bank along with four exotic accessions for comparative evaluation. Conserved germplasm have been evaluated for different morpho-physiological traits, antioxidants and nutrients, and under normal and moisture stress condition at seedling stage. Considerable diversity in vegetative parameters, fruiting traits, antioxidants, nutrients and response to moisture stress at seedling stage has been found. Moisture stress tolerant accessions have been found to show higher proline accumulation accompanied by higher activity of antioxidant enzymes. Untapped genetic variability available in this region may be utilized for cultivation as well as for crop improvement programmes.
PA092 (IPPC0668) Role of reactive oxygen species (ROS) during the growth of mungbean [Vigna radiata (L.) Wilczek] seedlings under water stress Satyajit Das* and Rup Kumar Kar Plant Physiology and Biochemistry Laboratory, Department of Botany, Visva-Bharati University, Santiniketan, West Bengal, India *Presenting author: [email protected]
Recently a role of ROS in growth and development in plants has been revealed. While working with Vigna radiata seedlings we observed that mild water stress (simulated by PEG 6000) promoted root growth while inhibiting growth in hypocotyl, which might be controlled by extracellular ROS. Thus, root growth of 3-day-old seedlings was retarded, both under water stress and non-stress condition, by treatment with CuCl2, a superoxide scavenger, as well as by ZnCl2, an inhibitor of NADPH oxidase (PM-located superoxide generator). However, in case of hypocotyl these treatments were without effect.
Treatments with DMTU (H2O2 scavenger) and DEDTC (SOD inhibitor), though inhibited normal growth, could not inhibit stress-induced root growth. Again these treatments did not effect hypocotyls. When seedlings were treated with sodium benzoate a OH scavenger, and SHAM, a peroxidase inhibitor, root growth under both stress and non-stress conditions was inhibited, will the former being more effective. Again these were not effective in hypocotyl. In-gel assay showed higher activity of NADPH oxidase (RBOH) in root, but not hypocotyl, under PEG-induced stress confirming the involvement of apoplastic superoxide in stress-induced root growth, although data on assessment of extracellular production of superoxide by estimating in bathing medium did not support this. But localization of superoxide and H2O2 by staining root and hypocotyl separately with NBT and TMB, respectively confirmed the involvement of ROS in water stress-induced growth promotion of root whereas these are not involved in growth inhibition of hypocotyl under water stress simulated by PEG 6000.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 160 Session A : Abiotic Stress Adaptation and Management
PA093 (IPPC0683) Identification of promising salt tolerant genotypes using stress indices of morphological and physiological trait at seedling stage in rice S.L. Krishnamurthy*, V. Batra, P. Pundir, P.C. Sharma and D.K. Sharma Division of Crop Improvement, Central Soil Salinity Research Institute, Karnal-132001, Haryana, India *Presenting author: [email protected]
A set of 131 rice accessions was evaluated in two levels namely, non-stress (EC ~ 1.2 dS/m) and saline stress (EC ~ 10 dS/m) in hydroponics at seedling stage. Root length and shoot lengths were reduced by 52 and 50%, respectively in saline stress in comparison to non stress conditions. The effect of the increased Na+ concentration in the medium is detrimental to root length and shoot length as observed by reduction in root length and a concomitant reduction in shoot length. The increased concentration of Na+ leads to augmented Na+/K+ ratio with increasing stress in medium and decreasing expression of traits. A significant positive correlation (r=0.60) was noticed between stress tolerance index (STI) for root length and STI for shoot length. The stress susceptibility index (SSI) for root length was expressed significant positive correlation with SSI for shoot length (r=0.43). SSI for K+ content was registered significant negative correlation with STI for Na+ content (r=-0.43). The three accessions namely, IC 545004, IC 545486 and IC 545215 were found to be the best performers adjudged on the morphological and physiological criteria in saline stress situation. These three rice accessions could be used as a donor parent or for genotypic studies in future breeding programs.
PA094 (IPPC0687) Development of salt tolerant varieties following conventional, physiological and molecular approaches Parbodh Sharma1*, S.L. Krishnamurthy1, Neeraj Kulshreshtha2, Jogendra Singh1, Preeti Pundir1 and D.K. Sharma1 1Central Soil Salinity Research Institute, Karnal-132001, Haryana, India 2Sugarcane Breeding Institute RRS, Karnal-132001, Haryana, India *Presenting author: [email protected]
Among abiotic stresses, salt stress is the most complicated problem posing a major challenge. Around 1000 m ha globally and 6.73 m ha in India are affected by salinity. Central Soil Salinity Research Institute (CSSRI) has developed 15 salt tolerant varieties of different crops; 7 of rice (CSR10, CSR13, CSR23, CSR27, CSR Basmati30, CSR36 and CSR43), 4 of wheat (KRL1-4, KRL 19, KRL210 and KRL213), 3 of Indian mustard (CS52, CS54 and CS56) and one in Chick pea (Karnal Chana 1) through conventional breeding approaches to increase the productivity of salt affected soils for resource poor farmers. Physiological and biochemical approaches have been used to identify different mechanisms of salt tolerance and used for identification of salt tolerant and sensitive lines. The molecular approaches are also being used to develop salt tolerant rice varieties. Introgression of Saltol QTL to two mega high yielding rice varieties namely, Pusa 44 and Sarjoo 52 carried out through Marker Assisted Selection Back Cross (MABC) breeding. The varieties namely, Pusa 44 and
Sarjoo 52 were selected as recurrent parents while FL478 was used as a donor parent and produce F1 seeds. True F1s were selected by the Saltol marker RM8094. Further backcrossing with respective recurrent parent and selfing was performed to produce seeds up to BC3F3 generation. In every back cross generation, recombinant and foreground selection was carried out. Desirable plants were selected based upon the (genotyping) marker data and through stringent phenotyping as similar to recurrent parent. Background selection and DUS characterization of NILs is in progress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 161 Session A : Abiotic Stress Adaptation and Management
PA095 (IPPC0688) Rice F-box protein, OsFBK, regulates abiotic stress signaling Satyam Vergish*, Nitin Jain and J.P. Khurana Department of Plant Molecular Biology, Interdisciplinary Centre for Plant Genomics, Delhi University, New Delhi-110021, India *Presenting author: [email protected]
The F-box proteins (FBPs) are components of SCF ubiquitin-ligase complexes and are one of largest multi-gene superfamily in plants containing a conserved N-terminal F-box motif that binds to the SKP, a component of SCF, and a variable C- terminal protein interaction domain that selectively binds target proteins for turn-over by 26S proteasome. There are over 900 FBPs in Arabidopsis and rice that are involved in all the facets of plant biology such as stress signalling, photomorphogenesis, hormone signaling, flower development, self incompatibility, biological rhythms, defence and so on. Our lab identified a few F-box proteins in rice that are regulated under abiotic stresses, one of which was OsFBK. The gene was found to be upregulated under drought, heat and salt stresses and also in Y-leaf and flag leaf. The protein harbors a canonical F-box domain which interacted with OsSKPs demonstrating that it is a component of a functional SCF-type E3 ligase. The OsFBK protein showed formation of homodimers and heterodimers with its paralogs in yeast two-hybrid assays. Also screening of heat stress rice library showed interaction of OsFBK with some proteins known to play roles in heat tolerance. The protein was found to localize in the nucleus as well as cytosol. Over-expression lines in Arabidopsis showed role of this gene in heat response and other abiotic stresses. Also, the over-expression lines flowered earlier. The above results indicate involvement of OsFBK in abiotic stress signalling and photoperiodic flowering.
PA096 (IPPC0693) Assessment of physio-biochemical traits of rice landraces of Goa for salt stress tolerance Kundan Kumar* and Swapnil Punyapwar Department of Biological Sciences, BITS Pilani K.K. Birla Goa Campus, Goa- 403726, India *Presenting author: [email protected]
Rice is the major staple food crop for more than half of the world’s population. Salinity stress is the most common and serious problem in rice cultivation mainly in the coastal saline soils of Goa locally known as Khazan land and occupies 33 percent of total rice cultivated area. Developing rice cultivars with tolerance to salt with better agronomic and quality traits acceptable to farmers is a feasible approach to address this problem. Salt tolerance trait available with certain traditional land races, not exploited so far, has significant relevance in crop improvement. The indigenous rice germplasm of Goa is rich in genetic diversity and valuable gene system for traits associated with salinity tolerance. In the present study, we have analyzed three rice genotypes of Goa namely Dhave, Ek Kadi and Korgut for their salt tolerance at seedling stage. IR64, a salt sensitive cultivar of Oryza sativa was used as a control. Their physiological parameters viz. root length, shoot length, fresh weight, dry weight, germination percentage were taken into consideration for salt stress tolerance. Biochemical parameters like salinity induced free proline accumulation, chlorophyll content, lipid peroxidation, catalase activity, total protein and elemental (Na+, Ca2+, K+) analysis were considered for salt stress tolerance in all 4 genotypes. Taken together our results suggest that Korgut cultivar showed better tolerance to salinity stress among all landraces tested here. The rice genotypes showing salinity stress tolerance will be highly useful in rice breeding for improved rice varieties to survive under higher salt stress condition.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 162 Session A : Abiotic Stress Adaptation and Management
PA097 (IPPC0699) Impact of ZnO nanoparticle on water stress and its recovery in Oryza sativa L. H. Upadhyaya1*, S. Shome1, M.K. Bhattacharya1, S. Tewari2 and S.K. Panda3 1Department of Botany and Biotechnology, 2Department of Physics, Karimganj College Karimganj-788710, Assam, India 3Plant Biochemistry and Molecular Biology Laboratory, School of Life Sciences, Assam, University, Silchar-788011, India *Presenting author: [email protected]
Zinc oxide (ZnO) nanoparticles (NP) is being widely used in various research. Plant system being extremely critical requires the fundamental understanding on the influence of ZnO-NP on its cellular growth and functions. Our study evaluates the effect of ZnO-NP on growth and physiological changes in Oryza sativa L., and its ameliorative effect on PEG induced water stress and its recovery in rice. Water stress induced growth reduction in rice seedlings, but the - ZnO- NP alleviatied the adverse effect of water stress on rice seedlings by stimulation of growth parameters and the increase in photosynthetic pigments. It can also be suggested that the water stress recovery may be counteracted by ZnO-NP at lower concentration. Further, detail study on the ZnO-NP effect on physiological events during water stress, and physiochemical and antioxidative responses of rice seedlings can reveal the exact mechanism underlying the amelioration of water stress and its recovery in rice. The present study gives an insight on the possible role of zinc transporter protein and their interaction with ZnO-NP during water stress acclimatization and stress recovery process in rice, which in turn will help in understanding biointerface on protein–nanoparticles interaction in future. Detail understanding of the regulation of zinc transporter in response to Zn nanoparticles is required to dissect the physiological mechanism regulated by ZnO- NP during water stress and its recovery in rice.
PA098 (IPPC0711) Validation of Sub1 QTL in CO 43 from submergence tolerance using key physiological traits R. Samundeswari1*, D. Vijayalakshmi1 and M. Raveendran2 1Department of Crop Physiology, 2Centre for Plant Molecular Biology, Tamil Nadu Agricultural University, Coimbatore-641003, India *Presenting author: [email protected]
In Tamil Nadu, Cauvery delta zone is frequently affected by flash flooding during the monsoon period. Hence, research was aimed towards improving submergence/flooding tolerance of popular rice genotypes of Tamil Nadu viz., CO43 by introgressing Sub1 locus from the tolerant FR13A through marker assisted selection. BC1F1 generation was forwarded up to BC4F1 and selfed progenies of BC2 and BC3 generations were phenotyped for submergence tolerance using key physiological traits. Advanced generations of the Sub1 introgressed CO43 lines were subjected to flooding stress 13 days in a simulated submergence screening facility and evaluated for traits namely leaf and stem carbohydrates, chlorophyll content, chlorophyll fluorescence (Fv/Fm), photosynthetic rates, stomatal conductance and transpiration rates. All the Sub1 introgressed lines survived 13 days of complete submergence showing more reserved carbohydrates before and after submergence, higher chlorophyll contents and chlorophyll fluorescence ratios coupled with high photosynthetic rates. CO43 could not survive 13 days of flooding and was found dead and decayed. Almost all the introgressed lines were found to be superior to FR13A, the donor parent. Superior progenies of CO43 improved for their submergence tolerance has been developed which will be further tested for their agronomic performance and efforts will be taken for release as a smart rice variety to submergence tolerance.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 163 Session A : Abiotic Stress Adaptation and Management
PA099 (IPPC0714) Marker assisted introgression of Sub1 locus from FR13A into CO43 for improved submergence tolerance D. Vijayalakshmi1*, M. Raveendran2, R. Samundeswari and S. Muthulakshmi 1Department of Crop Physiology, 2Central Institute for Cotton Research, Tamil Nadu Agricultural University, Coimbatore-641003, Tamil Nadu, India *Presenting author: [email protected]
The study was aimed at developing submergence tolerant version of popular rice variety of Tamil Nadu viz., CO43, through marker assisted introgression of Sub1 locus from a FR13A. BC1F1 generation was forwarded up to BC4F1 and selfed progenies of BC2 and BC3 generations were genotyped and phenotyped for submergence tolerance. Twenty one BC3F1 plants were subjected to BGS using 37 SSR markers. The effect of introgression of Sub1 locus in terms of tolerance against submergence was studied in BC2F3 and BC3F3 progenies under simulated submergence condition. CO43 plants were completely affected. All the survived plants in BC2F3 and BC3F3 generations were evaluated for plant height, carbohydrates, chlorophyll, chlorophyll fluorescence, photosynthetic rate, stomatal conductance and transpiration rates before and after flooding. All the introgressed lines proved to be flood resistant with improved physiological traits. Superior progenies of BC2F3 were selfed to generate BC2F4. BC2F4 progenies were raised in the field along with the parents and evaluated for the morphological traits. These superior Near Isogenic Lines (NIL’s) recorded the plant height and grain quality similar to that of CO43. Superior NIL’s of CO43 improved for their submergence tolerance have been developed which will be further multiplied and tested under multi-location trails.
PA100 (IPPC0725) Genetic engineering of RSA and source-sink relationship in rice for improving abiotic stress tolerance Preeti Kaur*, Pragya Yadav, Santosh Vinjamuri, Shashank Yadav, Rakesh Verma and Viswanathan Chinnusamy Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Rice requires about 5000 litres of water to produce 1 kg of grain. Drought stress is a major constrain for rice production in India. Phytohormones play significant roles in dehydration tolerance mechanisms of plants. A low level of cytokinin has been shown to promote root growth and drought tolerance in Arabidopsis and tobacco. Genetic and transgenic analyses in different plants revealed that cytokinin is vital for determination of grain number and maintenance of photosynthesis. This study was aimed at simultaneous improvement of root system architecture (RSA) and source-sink balance to enhance drought tolerance in rice. Rice plants were engineered to down regulate cytokinin levels in root by over-expression of cytokinin oxidase (CKX) by using root specific promoter (RCc3), and maintain appropriate levels of cytokinin in shoot by engineering drought responsive down-regulation of CKXs expression by RNAi in the shoot. Genes for cytokinin biosynthesis (IPT) and degradation (CKX) were identified from Oryza sativa genome. Selected genes were cloned into plant transformation vector. Transgenic rice of genotypes MTU1010 and IC305692 were developed using these constructs through Agrobacterium mediated transformation. Molecular analysis of transformants was carried out using PCR and RT-PCR. Rice transgenics will be further analyzed for altered RSA and source-sink relationship under normal and drought stress conditions. These transgenics may be useful in developing rice lines with improved RSA and enhanced yield stability under drought stress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 164 Session A : Abiotic Stress Adaptation and Management
PA101 (IPPC0741) Expression analysis of SOS pathway genes under salinity stress in contrasting rice genotypes T.V. Vineeth1*, Parbodh Sharma1, Ashwani Pareek2, Manjari Mishra2, Priyanka Das2 and S.L. Krishnamurthy2 1Crop Improvement Division, Central Soil Salinity Research Institute, Karnal-132001, Haryana, India 2School of Life Science, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Salinity is one of the major problems negatively influencing rice productivity. Salinity tolerance is a very complex trait. SOS (Salt Overly Sensitive) Pathway has been established as a major candidate in cellular ion homeostasis and thereby salt tolerance.The present study was carried out to examine the role of SOS pathway in salinity stress tolerance in selected rice genotypes. The experiment was conducted in hydroponics with 8 rice genotypes, viz., Pokkali, VSR 156, CSR 30, Pusa Basmati 1, Pusa 44, CSR 27, CSR 10 and IR 29 subjected to salinity stress (200 mM NaCl) along with control after 12 days of sowing. Imposed salinity treatment lead to significant reduction in relative water content (RWC) and membrane stability index (MSI) in all the genotypes. However, decline in RWC and MSI was significantly less in Pokkali, CSR 10 and CSR 27 as compared to other genotypes. In contrast, the sodium (Na) content increased under imposed salinity stress in the shoot tissues of all the genotypes. However, the increase was less in Pokkali, CSR 10 and CSR 27, which also maintained lower sodium/potassium (Na/K) ratio enabling cellular ion homeostasis. Gene expression analysis of CSR 10 and IR 29 revealed the presence of a more efficient salt overly sensitive pathway composed of SOS1, SOS2 and SOS3 in CSR 10 leading to a good correlation with the effective salinity stress tolerance of CSR 10 at the physiological level.
PA102 (IPPC0748) Analysis of sequence variation in HKT family genes of wheat for their association to salt tolerance Vijendra Singh*, Amit Kumar Singh, Rakesh Singh, Sundeep Kumar and Jyoti Kumari ICAR- National Bureau of Plant Genetic Resources, New Delhi-110012, India *Presenting author: [email protected]
Salinity is the major abiotic stress which limits wheat production in various parts of India and around the world. Some of the Indian wheat genotypes are known to possess very high level of tolerance to salinity stress. However, the molecular basis of salt tolerance in these genotypes is not clearly understood. A detailed understanding of molecular mechanisms underlying salt tolerance in these wheat genotypes will facilitate development of salt tolerant high yielding varieties. In the present study, a diverse set of 81 wheat genotypes including salt tolerant and salt sensitive genotypes were evaluated under controlled and stressed i.e., two different salinity conditions (4.0 and 5.0 dS m-1) in the net house at NBPGR, New Delhi. The leaf and shoot tissues from all the genotypes were harvested and analysed for Na/K concentration. Further, HKT (high affinity K transporter) genes have been amplified from wheat genotypes and analysed for the presence of sequence variations. Finally, nucleotide sequence variations among the HKT genes will be analysed for identification of SNPs/ Indels for their association with variation in level of Na/ K concentrations in the plant tissues of all the wheat genotypes.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 165 Session A : Abiotic Stress Adaptation and Management
PA103 (IPPC0762) Transgenic expression of cytosolic ascorbate peroxidase gene (apx1) from finger millet confers enhanced oxidative stress tolerance potential in Arabidopsis thaliana P. Yadukrishnan1*, Bhawna Negi2 and Sandeep Arora2 1Department of Biological Sciences, Indian Institute of Science Education and Research Bhopal- 462023, India 2Department of Molecular Biology and Genetic Engineering, G.B. Pant Univ. of Agriculture and Technology, Pantnagar-263145, India *Presenting author: [email protected]
A drastic increase in reactive oxygen species (ROS) level is associated with most of the biotic and abiotic stresses and attributes to the reduced crop productivity under stress. Halliwell-Asada pathway (Ascorbate-glutathione cycle) is a central pathway that effectively detoxifies hazardous ROS in plants. Ascorbate peroxidase (APX), an antioxidant enzyme plays a crucial role in this cycle by reducing toxic H2O2 to water using ascorbate as an electron donor. In this study, successful attempts were made to transform Arabidopsis thaliana (ecotype Col-0) with finger millet (Eleusine coracana) cytosolic ascorbate peroxidase (apx1) gene, cloned from a stress tolerant variety, PR202. Successfully transformed Arabidopsis seedlings carrying Ecapx1 driven by CaMV35S promoter showed increased net expression of ascorbate peroxidase. The functional validation of the transgenic plants for their ability to tolerate oxidative stress was done by carrying out quantitative estimation of biochemical parameters such as free proline accumulation, malondialdehyde (MDA) content and H2O2 content, under salinity-induced oxidative stress. Higher net accumulation of proline was observed in transformed plants when compared to wild types. Significant reduction in accumulation of MDA and H2O2 contents was recorded in transgenic plants as compared to the wild-type plants. The greater accumulation of proline and reduction in the toxic oxidative species contents indicate that transgenic Arabidopsis plants expressing apx1 from finger millet are better equipped to tolerate oxidative stress which could be well attributed to the presence of enhanced ascorbate peroxidase activity.
PA104 (IPPC0768) Mitigating effect of salicylic acid on biochemical and antioxidant enzymes in maize (Zea mays L.) genotypes under low temperature stress Preeti Singh* and Vijai Pandurangam Department of Plant Physiology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221005, U.P., India *Presenting author: [email protected]
An experiment was carried out in field condition to find out sowing effects along with salicylic acid treatments with two screened maize genotypes i.e., HUZM-185 (tolerant) and HUZM-80-1 (sensitive) under low temperature stress during Rabi season. Seeds were primed with salicylic acid (SA) @ 20 ppm and 40 ppm along with hydro (distilled water) for overnight and dry seeds as control before sowing in split plot design with three replications having two sowing condition i.e., normal and delay. Observations were recorded at 20, 40 and 60 (days) after sowing viz., malondialdehyde content
(MDA), hydrogen peroxide content (H2O2), superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX) activities in both normal and delay sowing. It was found that 20ppm salicylic acid significantly reduced MDA, H2O2 and enhanced SOD, CAT and POX activities in both genotypes on normal and delay sowing, but values were higher in delay sowing condition under low temperature stress. Remaining treatments were also performing better as compared to dry treatment under low temperature stress. Tolerant genotype significantly reduced MDA and H2O2 production and enhanced antioxidants (SOD, CAT and POX) levels as compared to sensitive genotype. Hydrogen peroxide is a reactive oxygen species (ROS), which leads to disarrangement of bio-membrane and hampers its selectivity. A higher antioxidant activity in tolerant genotype minimized ROS production (H2O2) and overcomes low temperature stress as compared to sensitive genotype. The deleterious effect of low temperature stress was managed by enhancing SOD, CAT and POX activities in delay sowing as compare to normal sowing condition.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 166 Session A : Abiotic Stress Adaptation and Management
PA105 (IPPC0769) Copper mediated effect on reactive oxygen species generation and antioxidant responses in Oryza sativa Nisha Kevat* and Prabhat K. Sharma Department of Botany, Goa University, Panjim-403206, Goa, India *Presenting author: [email protected]
The aim of the present study was to investigate the effect of copper (Cu) on external morphology, photosynthesis, photosynthetic pigment content, reactive oxygen species (ROS) generation and succeeding oxidative damage in rice plants (Oryza sativa L. var. Jaya). Isoforms of GPX (Guiacol peroxidase), CAT (Catalase), SOD (Superoxide dismutase) and SOD activity were also studied. Plants were grown in pots (12 cm diameter) containing vermiculite and irrigated with
0.1-3.0 mM CuSO4 in Hoagland’s solution. SEM images of the leaf surface revealed reduction in stomata and trichome size in a concentration dependent manner. Quantum efficiency of PSII (Fv/Fm), photochemical quenching (qP), net photosynthetic rate (A), transpiration rate (E) and stomatal conductance (gs) decreased radically with incriminate in Cu treatment. Chlorphyll a, chlorophyl b, neoxanthine, violaxanthine and antheraxanthin were reduced linearly with increasing
Cu concentration. Hydrogen peroxide (H2O2) and hydroxyl radical (OH•) were effectively generated with an elevated Cu concentration and induced oxidative stress measured as oxidative damage to lipid peroxidation and protein oxidation. Osmotic stress measured as proline content showed three-fold accumulation at 3 mM Cu concentration. Isozymes of GPX and SOD were greatly affected due to Cu stress. Pronounced enhancement in the SOD activity was observed to metabolize ROS. The results demonstrated that exposure to elevated Cu concentration resulted in ROS (H2O2 and OH•) generation leading to overall inhibition of photo-phosphorylation, brought morphological changes and induced oxidative stress. Eventually, the alleviated antioxidant responses emerged as major defensive mechanism adopted by rice plants to moderate the Cu stress.
PA106 (IPPC0770) Cloning and characterization of salt gene from Pusa Basmati 1 Navdeep Kaur*, Isha Sharma and Pratap Kumar Pati Division of Biotechnology, Guru Nanak Dev University Amritsar, Amritsar-143005, India *Presenting author: [email protected]
Salinity is one of the major stresses affecting rice production worldwide. It is fairly a complex process regulated by an array of genes. One such gene is SalT that is located in the Saltol QTL and shows higher expression under salt stress. However, precise mechanism through which it confers tolerance is not well understood. In the present work, attempts were made to extensively characterize this gene in rice. The expression of SalT was analyzed under different conditions using qRT-PCR and was found to be up-regulated in different tolerant varieties as the scale of tolerance increases. Further, it was found to be regulated by different hormones and range of abiotic stresses. SalT gene from rice was cloned and overexpressed in Nicotiana tobaccum for developing transgenic plants overexpressing rice SalT. These transgenic plants were analyzed for increased salt tolerance and for finding putative interactive partner using FLAG-pull down assay. Initial analysis showed that this gene could be regulated through MAPK pathway in a calcium dependent manner. The gene was further cloned into the protein expression vector and species specific antibody was developed. The antibody was used for the validation of gene expression at the protein level. The heterologously expressed purified protein was used for protein structuaral characterization using SAXS and was found to exist in different isoforms, dimeric form being most stable. The protein was observed to bind specifically with mannose that was confirmed using ITC and gel retardation assay indicating the probable link between carbohydrate metabolism and SalT.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 167 Session A : Abiotic Stress Adaptation and Management
PA107 (IPPC0771) Physiological characterization of mungbean genotypes for salinity stress tolerance Sunder Singh1*, Neeraj Kumar2 and A.S. Nandwal3 1Department of Botany, Maharshi Dayanand University, Rohtak-124001, Haryana, India 2Department of Botany and Plant Physiology, CCS Haryana Agricultural University, Hisar-125004, Haryana, India *Presenting author: [email protected]
Two mungbean genotypes (Asha and Muskan) were evaluated for salinity stress tolerance on the basis of physiological traits viz. plant water status, solute accumulation, ionic composition, reproductive behavior and grain yield. Both the genotypes were raised in green house and treated with salinity stress of 0, 2.5, 5.0 and 7.5 dS m-1 at flowering stage for ψ ψ ten days and analysed for above traits. The water potential ( w) of leaves and osmotic potential ( s) of leaves and decreased with increase in salinity levels from 0 to 7.5 dS m-1 in both the genotypes. Relative water content of leaves and roots reduced, while proline and total soluble sugar (TSS) increased under salinity stress. The Na+/K+ ratio in leaves and roots enhanced significantly and highest was estimated in roots. The genotype ‘Asha’ exhibited a low Na+/K+ ratio in aerial plant parts compared to ‘Muskan’. Salinity stress delayed the time of first flowering initiation by 2-4 days and reduced the number of flowers per plant. Pollen viability and in vitro pollen germination decreased in both the genotypes due to salinity. Similarly, the pollen tube length decreased with increasing the level of salinity but did not effect on the number of ovule production. Salinity stress reduced the number of pods, number of seeds and weight of seeds per plant; and seed test weight. Based upon above physiological traits, the genotype ‘Asha’ was identified relatively tolerant to salinity stress than ‘Muskan’.
PA108 (IPPC0802) Effect of water stress on physiological, biochemical, yield and yield contributing components in pigeon pea (Cajanus cajan L. Millsp.) Damor Parthvee R1, Ajay V. Narwade2*, Lalit Mahatma3, D.A. Chauhan4 and S. Sree Ganesh5 1,2,5Department of Genetics & Plant Breeding, 3Department of Plant Pathology, 4Main Pulses and Castor Research Station, N.M. College of Agriculture, Navsari Agriculture University, Navsari-396450, Gujarat, India *Presenting Author: [email protected]
A field experiment was conducted during winter (rabi) season of 2013 at Navsari Agricultural University, Navsari, to evaluate the effect of water stress on physiological, biochemical, yield and yield contributing components in pigeon pea
(Cajanus cajan L. Millsp.). Four irrigations treatments were given during the experiment, viz., I0- all the irrigations given at
25, 50 and 75 days after sowing (DAS), I1- Irrigation given at 25 and 50 DAS, I2- Irrigation given at 25 DAS and I3- kept as rainfed. Six varieties of pigeon pea were used, viz., GT-102, Bharboot local, GNP-304, GT-1, AGT-2 and C-11 in split plot design with three replications. The response of pigeon pea varieties were observed by measuring various physiological, biochemical, growth, yield and yield attributing parameters. The experimental results indicated that the seed yield of pigeon pea was significantly lower due to water deficit, due to the reduction in various physiological, biochemical, growth, yield and yield attributing characters. Among the six pigeon pea genotypes, GT-102 recorded significantly highest yield as compared to other varieties due to highest harvest index, number of pods per plant, number of seeds per pod, 100- test weight, seed yield per plant, leaf area index, leaf area ratio and photosynthetic rate. It also had the highest accumulation of chlorophyll, carotenoid, glycine-betaine, proline and protein. By these results, GT-102 was identified as the drought tolerant variety, followed by C-11, GT-1 and AGT-2, while two the other genotypes, GNP-304 and Bharboot local were drought susceptible.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 168 Session A : Abiotic Stress Adaptation and Management
PA109 (IPPC0806)
Photosynthesis and yield of chickpea (Cicer arietinum) under interactive effect of elevated CO2, drought and high temperature Sadhana Maurya1,2*, Ashish K. Chaturvedi1, Shashi Meena1, Madan Pal1 and A.K. Chopra2 1Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi- 110012, India 2Department of Zoology and Environmental Science, Gurukula Kangri Vishwavidyalaya, Haridwar-249404, Uttarakhand, India *Presenting author: [email protected]
India, the largest producer of pulses in the world, shares 14.6% of global pulse production. Chickpea has been known sensitive to drought and high temperature which causes reductions in yield. On the other hand, CO2 enrichment studies revealed beneficial effects of CO2 in terms of photosynthesis and yield in chickpea. However, combination experiments on elevated CO2, high temperature and drought are meagre. Therefore, an attempt was made to determine the potential impact of elevated CO2, drought and high temperature alone and in combination of all the factors on photosynthesis and seed yield of two contrasting chickpea genotypes viz. ICC-4958 and FLIP 90-166 raised under open top chambers. Pot- grown plants grown inside open top chambers were exposed to elevated CO2 (ambient +250 μl/l), high temperature (>5°C above ambient) and drought conditions (upto -1.6MPa) throughout the growth season. Elevated CO2 alone significantly increased photosynthetic rate, plant biomass and seed yield of both the genotypes while high temperature and drought alone caused significant reduction in photosynthetic rate and contributed to poor vegetative growth with lesser biomass and seed yield compared to control. Elevated CO2 in combination to drought and high temperature as well showed ameliorative effects on both the genotypes, however, the response was more prominent in FLIP 90-166. This study concludes that elevated CO2 can alleviate the adverse effect of drought and high temperature stress on photosynthetic C assimilation and yield in chickpea.
PA110 (IPPC0809) Evaluation of maize genotypes for cold tolerance Hari Singh Meena1, Upama Mishra1*, R.N. Gadag1 and H. Pathak2 1Division of Genetics, 2CESCRA, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Cold stress is a major environmental factor that limits the agricultural productivity of maize plants. Though maize is a major crop in kharif season, cultivation during winter season in the Indian sub-continent is assuming importance due to various factors. Low temperature often affects plant growth and crop productivity, which causes major loss in maize crop in the northern part of India. Identification of suitable genotypes under northern belt of the country, therefore, assumes priority, to cater to such agro-climatic conditions. About 1300 lines of maize, in various stages of their derivation and diverse sources were screened against cold stress in field conditions. Observation recorded were days to germination, days of silking, days of anther shedding, stem colour, root colour, anther colour, silk colour, uniformity, vigour, no of cobs, height. Days to germination ranged from 25 days to 41 days after sowing. Days of silking ranged from 84 days to 112 days. Extensive variability was found in stem colour, ranging from light yellow to dark green including purple colour. Secondary roots colour ranged from light green to dark green including purple. Silk colours were brown and red. Anthers colour in the maize genotypes ranged from red brown to blue. Based on these observations and taking into account the productivity of the lines in the experimental field of IARI, 52 lines which performed relatively better under cold stress, were identified. These selected lines will be subjected to re-screening in next rabi season and also would be involved in generating hybrids specifically suitable for rabi season in the Northern part of India. This strategy is expected to make use of wide flexibility and diversity of maize in terms of adaptation to varied climatic conditions and could serve as additional, economically viable option for the farmers.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 169 Session A : Abiotic Stress Adaptation and Management
PA111 (IPPC0810) Fatty acids composition of fenugreek (Trigonella Foenum-graecum L.) seed oil as influenced by water stress S.N. Saxena*, R.K. Kakani and S.S. Rathore Department of Plant Physiology, National Research Centre for Seed Spices, NRCSS, Tabiji Farm-Ajmer 305206, Rajasthan, India *Presenting author: [email protected]
A study was conducted with thirteen fenugreek genotypes to evaluate the effect of water stress on oil content and its fatty acids composition. Oil content under non stress environment was ranging from a minimum of 2.62% in genotype AFg 6 to a maximum of 5.33 in AM 327-3. When stress imposed at midterm growth stage the oil content was increased from a minimum of 3.29% in AFg 6 to a maximum of 5.31 in AM 327-3. Significant genotypic variation was observed in fatty acids composition. Linoleic acid (34-54%), linolenic acid (27-46%), stearic acid (0- 5%) and palmitic acid methyl ester (7-13%) were major contributors. Genotypes AFg 3, AFg 5, B2 19, and CL 32- 17 showed increase in lenoleic acid and either decrease or no change in lenolenic acid under stress conditions while genotypes AFg 4, AFg 6, AM 292, AM 327-3, Hisar Sonali, AL-1-2 and RMt 305 showed decrease in linoleic acid and increase in lenolenic acid content. Stearic acid and palmitic acid methyl esters either reduced or unaffected under stress conditions. Under non stress genotype AFg 3 gave maximum seed yield (8.93 g plant-1). Genotype AFg 6 produced highest seed yield (8.99 g plant-1) under stress conditions where lenoleic acid drastically reduced from 49.57 to 34.85% and lenolenic acid increased from 31.69 to 37.66%. Other genotypes viz. AFg 4, AFg 6, Hisar Sonali and RMt 305 yielded good under stress showed similar fatty acid composition.
PA112 (IPPC0823) Improving salt tolerance of crop plants : prospects and perspectives Surinder Kumar Sharma* Department of Crop Improvement, ICAR-Central Soil Salinity Research Institute, Karnal-132001, Haryana, India *Presenting author: [email protected]
More than 800 million hectares of land throughout the world and 6.73 million hectares in India are salt-affected and / or accompanied by poor quality waters. Despite the increasing severity of these problems and their detrimental effects on agricultural production, programs to improve plant tolerance are in place in few countries, national and international research organizations. Selection for tolerance in the saline / sodic soils is further complicated as soil heterogeneity is often accompanied by other individual or multiple stresses like drought, waterlogging, mineral deficiencies or toxicities. Four decades of researches at CSSRI, Karnal involving conventional breeding techniques supplemented by physiological and genetic mechanisms helped to develop suitable materials culminating in release of 15 salt tolerant varieties in rice, wheat, Indian mustard and chickpea besides identifying tolerant genetic stocks and donors. Some of these materials also possess multiple tolerance to drought, waterlogging or micro-elements enabling better performance and adaptability in diverse stress situations. These efforts were supported by demonstrations in research farms and farmer’s fields, supplemented by production and supply of quality seed promoting their cultivation and spread in many states having salt affected lands. Adoption of these varieties has made significant contribution to national food production and poverty alleviation, benefiting resource poor and marginal farmers and restoring the marginally productive / barren lands into the production chain with additional environmental benefits. This paper describes some of these endeavors, adaptation and spread of such varieties in India and other countries highlighting the need to strengthen such programs globally.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 170 Session A : Abiotic Stress Adaptation and Management
PA113 (IPPC0836) Physiological and molecular analysis of root system architecture under drought and osmotic stress in rice G.K. Krishna1*, C. Vishwakarma, S. Thomas, P. Yadav, S. Srivastava, S. Yadav, R.K. Verma, V.V. Santosh Kumar, M. Das and V. Chinnusamy ICAR-Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Rice, the staple food for more than half the world population, is the second most important food grain crop. According to FAO, rice requires 1000 litres of water to produce 1 kg grain. Hence, reduced availability of irrigation water significantly affect its production. We targeted the avoidance mechanism exhibited by rice ie., the improved root system architecture (RSA) under drought stress. In the present study, 50 genotypes of rice were screened for their performance in RSA traits. Plants were raised in field, soil filled PVC pipes and hydroponic culture. In soil, treatment was given by cycles of drought stress and recovery at -50 and -10 kPa respectively. Under hydroponics culture, osmotic stress was given at -100 kPa by 8% PEG. From screening experiments, three contrasting genotypes were identified for RSA under stress viz. IC 305692 (better performing), Nagina22 (moderate) and RCPL 1-3c (poor). Further, to understand the genetic basis for the regulation of this complex trait, 29 candidate genes which were induced in roots under drought were selected from rice oligonucleotide array database. RT-PCR was performed to study their genotypic and tissue specific gene expression under 8% PEG and 400 nM ABA. Five genes were identified by this experiment - homeobox associated leucine zipper, universal stress protein, OsFBX132, MYB transcription factor and protein phosphatase 2C. The genes were subsequently cloned from rice, to be used for functional validation in Arabidopsis. The present study will provide light in to the mechanisms regulating root system architecture in rice, especially under drought stress.
PA114 (IPPC0837) Oxidative stress response in Lycopersicon esculentum mill. exposed to malathion and endosulfan Manisha Gautam1, Shefali Poonia1* and Purushottam2 1Department of Botany, Deva Nagri College, Meerut-250001, India 2Sardar Vallabh Bhai Patel University of Agriculture & Technology, Modipuram, Meerut-250110, India *Presenting author: [email protected]
A study was conducted with two varieties of tomato, viz. Pusa Ruby (PR) and Pusa Early Dwarf (PED). The plants were exposed to three different concentrations (0.05, 0.15 & 0.25%) of pesticides namely malathion and endosulfan at regular intervals. Proline and ascorbic acid content in leaves were estimated for oxidative stress response. There was a high accumulation of the two biochemical compounds which was concentration dependent. A substantial percent increase could be observed in proline content upto 22.22 and 25.21 and ascorbic acid content upto 22.52 and 43.12 with malathion and endosulfan respectively at plant age of 30d in PR. For the same in PED, proline was 12.41 and 19.71 while ascorbic acid was 28.31 and 34.04, which is comparatively less than PR. The two biochemical components exhibited an increasing trend with the age of the plant which slowed towards late maturity. At 90d, there was a significant decline in the increasing percent values of ascorbic acid both in PR and PED. Similar was the observation for proline in PR but in PED the increasing trend was observed. The enhanced accumulation of proline and ascorbic acid may be supportive to both the varieties of tomato plant. All the values obtained were subjected to statistical analysis and level of significance was evaluated.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 171 Session A : Abiotic Stress Adaptation and Management
PA115 (IPPC0843) Role of advanced unsaturated soil mechanics in understanding soil-root-water interaction V.K. Gadi1*, A. Garg1, S. Sreedeep1, W.H. Zhou2 and L. Sahoo3 1Department of Civil Engineering, 3Department of Biotechnical Engineering, Indian Institute of Technology, Guwahati-781039, India 2Department of Civil and Environmental Engineering, University of Macau, India *Presenting author: [email protected]
Fundamental understanding of soil-root-water interaction is important for analyzing and design of plantation scheme, irrigation scheduling and also in geotechnical infrastructures such as green roof, biofiltration units and vegetated slopes/ landfill covers. Unsaturated soil parameters such as soil water retention curve and hydraulic conductivity could be affected by soil type (particle size distribution), soil density and root growth (root depth and root distribution). The objective of this study is to critically review literature and highlight the short comings in terms of measurement of unsaturated soil parameters. A wide literature focusing on plant biotechnology and geotechnical discipline is reviewed. In most of the cases, parameters are burrowed from literature. On the other hand, literature in geotechnical discipline lacks studies on investigation of effects of root growth on unsaturated soil parameters. In addition, in their studies, the boundary condition (i.e., evaporation) at surface of ground is considered to be uniform without considering canopy effects of vegetation. The review suggests the importance of integration of advanced concepts of unsaturated soil mechanics with that of plant biotechnology for thorough understanding of soil-root-water interaction.
PA116 (IPPC0850) Comparative phenotyping under pot and field conditions to decipher component traits of drought tolerance in rice Rakesh Verma, Chandapal Vishwakarma*, Sitaram Kushwaha, Suchitra Pushkar, Shashank Yadav, Rahul Tiwari, Santosh Vinjamuri and Viswanathan Chinnusamy Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Genetic improvement of rice for enhanced WUE and drought tolerance is necessary to meet the food demand under declining fresh water availability. Evaluation of germplasm and breeding populations in augmented design for drought tolerance under field conditions often faces limitations due to soil moisture variability across the field and phenological differences among genotypes. Dehydration tolerance traits are poorly utilized in genetic improvement. Under limited soil conditions as in pots, mechanisms of transpiration minimization and cellular dehydration tolerance help survival of plants under drought stress. Hence, a comparative analysis of performance of rice genotypes was carried out for drought stress tolerance under pot and field conditions. Drought stress was imposed at flowering stage. Soil moisture status was quantified by tensiometer, soil moisture meter and gravimetric methods, while relative water content was measured to quantify plant water status. Various component traits such as tissue temperature, excised leaf water loss, chlorophyll stability index, spikelet fertility and yield components were measured. MSAP analysis revealed genotypic differences in drought induced changes in DNA methylation levels and methylated sites between drought tolerant rice genotype Nagina 22 and relatively drought sensitive genotype Swarna. Significant variation in dehydration avoidance and tolerance traits were found among genotypes. The results suggest that combined use of both pot and field methods will help greatly to tap the unexploited gene pool for cellular tolerance traits to enhance the yield of rice under drought stress environments.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 172 Session A : Abiotic Stress Adaptation and Management
PA117 (IPPC0856) Evaluation and validation of Saltol QTL for seedling stage salinity tolerance in rice (Oryza sativa L.) K.T. Ravikiran*, S.L. Krishnamurthy and Parbodh Sharma Genetics and Plant Breeding, Central Soil Salinity Research Institute, Karnal-132001, Haryana, India *Presenting author: [email protected]
The present investigation was undertaken to evaluate a set of 192 rice genotypes using hydroponics under saline stress (EC ~ 12 dS/m) and non stress (EC ~1.0 dS/m) conditions for morpho-physiological traits. Shoot length, root length and chlorophyll content decreased significantly under salinity as compared to non stress. Vigour score ranged from 3 (tolerant) to 9 (highly susceptible) with 14 and 16 genotypes categorized as tolerant (score 3) and moderately tolerant (score 5), respectively. The genotypes were further assessed for their molecular diversity using 10 SALTOL associated markers. A total of 30 alleles were generated at 10 microsatellite loci among the 192 genotypes. Combining morphological and molecular assessment, four genotypes i.e. CSR IR 34, CST 7-1, Arvattelu and IR 87938-1-1-3-1-2-B were found tolerant with distinctive allelic pattern from the tolerant check (FL 478). These could be used for further study to confirm their novelty and future breeding programme for seedling stage salinity tolerance in rice.
PA118 (IPPC0863) Insights into various mechanisms adopted by Arbuscular mycorrhizal (AM) symbiosis for sustainable legumes production under salt and metal(loid)s stresses Garg Neera* Department of Botany, Panjab University, Chandigarh-160014, India *Presenting author: [email protected]
Chickpea and pigeonpea are sensitive to salt and metal(loid)s stresses. Increased level of stresses negatively influenced germination, rhizobial symbiosis, photosynthetic efficiency, membrane properties, nutrient and redox homeostasis, osmoregulation; which eventually led to reduced plants growth and reproducibility. In pigeonpea, presence of high concentration of zinc counteracted the negative effects of Cd, while co-occurrence of Cd-Pb and Cd-NaCl in rhizosphere displayed synergism in the form of compound negative effects on growth and metabolism. In addition to rhizobial symbiosis, legumes can establish dynamic interaction with arbuscular mycorrhizal (AM) fungi, which is an essential link for effective phosphorus (P) nutrition, leading to enhanced N2 fixation. The mechanisms adopted by AM in alleviating salt and metal(loid)s stress included higher water absorption capacity and nutrient status by increasing root rhizosphere; enhanced nitrogen fixation; improved ionic and redox homeostasis. Besides this, mycorrhization reduced negative impact of metal(loid)s through dilution effect and phytochelatins induced vacuolar sequestration. Among different AM species, exotic single isolates conferred higher salt tolerance to pigeonpea than the native inoculums isolated from saline soils. Moreover, Rhizophagus irregularis (alone or in combination with Funneliformis mosseae) displayed higher effectiveness correlated with their more stable viability under salt stress. Although, abiotic stresses reduced AM colonization in both legume species; application of exogenous alleviants (flavonoids and silicon) further enhanced salt tolerance in chickpea through improved mycorrhization. Individually F. mosseae inoculation outperformed flavonoid and silicon application in imparting salt stress tolerance, however functional complementarity of F. mosseae with both flavonoids and silicon provided utmost yield and productivity in chickpea.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 173 Session A : Abiotic Stress Adaptation and Management
PA119 (IPPC0868) Screening of rice genotypes for heat tolerance based on chlorophyll fluorescence measurement N. Veronica1, Desiraju Subrahmanyam2*, S.R. Voleti2, Y Ashoka Rani1, K.L. Narasimha Rao1, M Lal Ahamed1 and P. Prasuna Rani1 1Agricultural College, Acharya N.G. Ranga Agricultural University, Bapatla-522101, India 2ICAR-Indian Institute of Rice Research, Rajendranagar, Hyderabad-500030, India *Presenting author: [email protected]
In the present scenario of climate change, high temperature has become a major concern challenging the productivity of rice. Hence identification of tolerant genotypes is pre-requisite to mitigate the adverse effects of climate change and global warming. Chlorophyll fluorescence traits gives insights into the ability of a plant to tolerate environmental stresses. Fv/Fm ratio, representing the primary photochemical efficiency of PSII is used as a selection criterion under heat stress condition. A diverse set of 58 rice genotypes comprising of 2 germplasm lines, 11 green super rice lines, 2 introgressed lines, 8 landraces, 2 tropical japonicas and 33 released varieties were screened based on their maximum quantum efficiency of PSII (or PSII photochemistry), measured by Fv/Fm values. Flag leaves at flowering stage were collected. One set was kept at ambient temperature (control) and another was subjected to high temperature in a growth chamber maintained at a temperature of 420C and light intensity of 300 μmol m-2 s-1 for 2 hours. Florescence measurements were made using a portable PAM Fluorometer (Walz PAM-210). Exposure to high temperature (420C) resulted in significant reduction in mean Fv/Fm ratio in all the tested genotypes. Significant variation was noticed amongst the genotypes in per cent reduction in Fv/Fm. The reduction was less than 5% in Assanchidiya, GSR 330, Khudaridhan, Akshayadhan and Rasi, indicating these genotypes are relatively tolerant to high temperature stress. On the contrary, more than 35% reductions were noted in Mahamaya, MTU 1001, RNR 6378 and Pantdhan 18, suggesting that the photosynthetic apparatus of these varieties were sensitive to high temperature stress.
PA120 (IPPC0871) Identification and characterization of heat- stress tolerance in maize (Zea mays L.) Suprokash Debnath1, Anuradha Gautam2, Pranjal Yadava2, Madan Pal1 and Ishwar Singh2* 1Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, Pusa, New Delhi-110012, India 2ICAR-Indian Institute of Maize Research, Pusa Campus, New Delhi-110012, India *Presenting author: [email protected]
Among the ever-changing components of the environment, high temperature is one of the most detrimental stresses for maize. Therefore, promising parental lines (inbred) with adaptive traits of tolerance to high temperature needs to be identified. In the present study, we evaluated 20 promising maize inbred lines of diverse genetic background for their per se performance at high temperature (summer season) as compared to control (kharif season), based on the different heat-stress adaptive traits like leaf area index, leaf firing, tassel blasting, anthesis-silking-interval and grain yield, The performance of DTPYC9F119 was best among all the genotypes under heat-stress, while K64R was found to be highly susceptible to high temperature. These contrasting genotypes were further analyzed under 6 days heat treatment (38/ 28°C) at flowering stage under managed stress conditions to understand the mechanisms of heat stress tolerance. Chlorophyll a and b ratio under heat-stress increased significantly in DTPYC9F119 after 6 days high temperature treatment. Net photosynthetic rate reduced under high temperature in K64R but the reduction rate was relatively lower in case of DTPYC9F119. Stomatal conductance increased under stress treatment in both the genotypes but the rate of increase was lower in DTPYC9F119. Activity of anti-oxidant enzymes (catalase, peroxidase and superoxide dismutase) and their genes showed differential expression in the two contrasting genotypes. Therefore, it seems that the tolerant genotype has evolved adaptive strategies by modulation of anti-oxidant gene expression to tolerate high temperature.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 174 Session A : Abiotic Stress Adaptation and Management
PA121 (IPPC0891) PGPR- mediated modulation of proline metabolic gene expression in Arabidopsis thaliana under drought stress Daipayan Ghosh* and Sridev Mohapatra Biological Sciences, Birla Institute of Technology and Sciences Pilani Hyderabad Campus, Shamirpet, Hyderabad-500078, India *Presenting author: [email protected]
Plant growth promoting rhizobacteria (PGPR) is a group of soil bacteria that are well known to contribute positively towards alleviation of abiotic stress in plants. Although some of the physiological processes involved in plant-PGPR interaction for stress alleviation are known, the precise molecular mechanisms still remain unclear. One of the mechanisms by which plants abate drought stress is through osmotic adjustment, by accumulating several compatible solutes, such as the amino acid proline. Here we report novel observations on the modulation of proline metabolic genes under dehydration conditions in the model plant Arabidopsis thaliana, by a beneficial rhizobacteria, Pseudomonas putida GAP-P45. Arabidopsis thaliana plants inoculated with Pseudomonas putida GAP-P45 under dehydration conditions exhibited significant delay in stress senescence with diminished symptoms of dehydration such as chlorosis, growth stunting and necrosis. Post drought induction, this particular strain of PGPR was capable of maintaining overall health of plants as opposed to the non-inoculated plants as concluded through visual and comparative physiological studies (analysis of fresh weight, dry weight, plant water content, total proline content etc.) estimated at different time points. Differences in root structure architecture were observed in the inoculated vs. non-inoculated plants under drought conditions. Semi-quantitative and quantitative real-time expression analysis of proline metabolic genes indicated a direct upregulation of proline biosynthetic genes and steady catabolism rate maintenance in plants by PGPR for long term survival under drought stress. We conclude that this strain of PGPR helps in drought mitigation of A. thaliana by modulating gene expression in the proline metabolic pathway.
PA122 (IPPC0897) Arsenic induced phytotoxicity in black gram (Vigna mungo L.) during early growth stage M.Z. Shamim* and A. Pandey Department of Biotechnology, Motilal Nehru National Institute of Technology Allahabad, Allahabad-211004, U.P., India *Presenting author: [email protected]
Arsenic is known as one of the highly toxic element to biological systems. Present study was designed to investigate the phytotoxic effects of arsenic (As) on different physiological parameter in black gram. Two genotypes of black gram, i.e., IPU 99-176 and UPU 83-35 were used for the experiment. Plants were irrigated with Hoagland’s hydroponic nutrient solution. Fifteen days after sowing plants were treated with water (control), 50 and 100 μM sodium arsenite along with Hoagland’s solution. The increasing arsenic stress condition significantly decreased root length (8.66-24.41 %), fresh root weight (7.61-89.36 %), shoot length (14.63-25.37 %) and fresh shoot weight (13.73 -26.56 %) with respect to control. At 100 μM As reduction in total chlorophyll was 12.91 and 20.4 5% in IPU 99-176 and UPU 83-35, respectively, with respect to control. Amount of chlorophyll-a increased (15.49 %) with increasing concentration of As in IPU 99-176, whereas it decreased in UPU 83-35 under increasing concentration of As. The amount of chlorophyll-b drastically decreased at 50 μM As concentration, but at 100 μM concentration the amount of chlorophyll-b increased with respect to 50 μM As concentration in both the genotypes. Carotenoids contentt in both of the genotypes decreased, and reduction was greater at 50 μM As concentration than 100 μM As concentration. Results indicated that IPU 99-176 black gram genotype may have greater tolerance for arsenic toxicity.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 175 Session A : Abiotic Stress Adaptation and Management
PA123 (IPPC0911) Epiphytic orchids of Assam and their adaptation to various abiotic stresses Ranjan Kumar Bora*, Nabami Basumatary and C.M. Sarma Department of Botany, Pragjyotish College, Guwahati-781009, Assam, India *Presenting author: [email protected]
Assam, a biodiversity rich state of India, harbours 290-odd species of orchids and of which majority are epiphytic. In nature, epiphytic orchids can survive and grow in minimal environments withstanding various abiotic stresses such as drought, temperature, light, nutrients etc. A study was conducted on epiphytic orchids of Assam to understand their mode of adaptation to different abiotic stresses during 2005 to 2014. Epiphytic orchids survive in minimal nutrient availability. During rainy season orchid roots absorb dissolved nutrients through stem flow and for which retention of water for longer period is necessary. To get better accumulation, epiphytic orchids adopt the following modes such as formation of a cavity by the root system, extending long roots to great distance, forming patch, encircling the stems and accumulation of debris by upright roots. Though orchids prefer diffused sunlight, the light requirement varies species to species and prefers to establish on the phorophyte according to their requirement. Shade loving orchids usually prefer to grow on barks near the ground. Tiny orchids usually prefer small twigs under the canopy. High light tolerant species have the ability to increase the carotenoids which protect the chlorophyll content from photo oxidation. Many epiphytic orchids have terete and scale leaves to withstand against extreme drought and high temperature. For better management of water, orchids possibly open their stomata during the early morning and in the evening. Otherwise, epiphytic orchids may act as CAM cycling physiotype during severe drought which helps the uptake of CO2 and conservation of water efficiently.
PA124 (IPPC0916) Abiotic stress mediated modulation of chromatin landscape in A. thaliana Vivek Kumar Raxwal1, Sourav Ghosh2, Somya Sinha1, Surekha Katiyar-Agarwal3, Shailendra Goel1, Arun Jagannath1, Amar Kumar1, Vinod Sacaria4 and Manu Agarwal1* 1Department of Botany, University of Delhi, New Delhi-110007, India 2Genomics and Molecular Medicine, CSIR-Institute of Genomics and Integrative Biology, Mathura Road, New Delhi, India 3Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India 4GN Ramachandran Knowledge Centre for Genome Informatics, CSIR-Institute of Genomics and Integrative, New Delhi, India *Presenting author: [email protected]
Considerable progress has been made in understanding abiotic stress-mediated physiological responses and their correlation with changes in candidate gene expression patterns. With significant advancements in genomic technologies, it is now possible to study transcriptional regulation in response to abiotic stresses on a genome-wide scale. One of the major constraints to transcription is the packaging of DNA by histones, which hinders accessibility of regulatory proteins to cis-elements. During transcription, chromatin dissociates the bound histones, which is later repackaged on completion of the transcriptional process. Hence, at any given stage, the chromatin is in a dynamic state w.r.t. its association with histones. We employed two different cross-validating methodologies, DNase-seq and FAIRE-seq (Formaldehyde Isolation of Regulatory Elements) to capture the open chromatin regions in A. thaliana genome upon imposition of four different abiotic stresses. Open chromatin regions identified in both FAIRE-seq and DNase-seq (OCFD) samples were used to analyze chromatin states relative to the control samples. Three categories of relative chromatin states were defined viz., (1) OC-OS in which chromatin was open in both control as well as stress samples (2) OC-CS, which included chromatin regions that were open in control and closed in stress samples and (3) CC-OS having chromatin regions that were closed in control and open in stress samples. Identification of relative chromatin states led to the elucidation of stress specific categorization of chromatin conformation. In conjunction with mRNA-seq data, this study allowed genome-wide identification of genes that are significantly up-regulated or down-regulated in response to different abiotic stresses.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 176 Session A : Abiotic Stress Adaptation and Management
PA125 (IPPC0931) A twist in the “plant- rhizobacterial interaction” tale in drought mitigation Diti Shah1, Sunetra Sen1*, Sridev Mohapatra1, Minakshi Grover2, Shalini Akula3 and Daipayan Ghosh1 1Biological Sciences, Birla Institute of Technology and Sciences Pilani Hyderabad Campus, Shamirpet, Hyderabad-500078, India 2Central Research Institute for Dryland Agriculture, Santoshnagar, Saidabad, Hyderabad-500059, India 3J.K. Agri- Genetics Ltd., Hyderabad, India *Presenting author: [email protected]
Plant growth promoting rhizobacteria (PGPR) have been known to contribute positively towards abiotic stress mitigation in plants. It is known that several strains of PGPR secrete plant growth promoting chemicals such as phytohormones which may help in modulating plant responses to abiotic stresses. Here, we report some novel observations in context of the interaction between the model plant, Arabidopsis thaliana and a Pseudomonas putida strain of PGPR which releases copious amounts of the auxin, indole acetic acid (IAA) during drought stress. Contrary to previous studies on PGPR, this particular strain, on inoculation with A. thaliana under drought conditions, exhibited a strong inhibitory effect on drought tolerance ability of the plant, while promoting plant growth under well-watered conditions. Inoculation of A. thaliana with this strain of bacteria under dehydration conditions reduced the plant’s inherent ability of transient drought mitigation. Plants exposed to drought with simultaneous P. putida inoculation exhibited enhanced drying, reduced plant water content, root growth inhibition and an overall attrition in drought tolerance as opposed to drought induced plants without PGPR inoculation. We conclude that this strain of P. putida (in the concentration used in the present study), enhances the susceptibility of A. thaliana to dehydration stress. We hypothesize that excess amounts of secretory/excretory IAA may be contributing to the negative impact of this strain towards drought mitigation in A. thaliana.
PA126 (IPPC0964) High temperature tolerance in chickpea genotypes as evaluated by photosynthetic rate, SPAD, PS-II, canopy temperature depression and yield stability index Pattipaka Bhasker1, Ajit Singh Nandwal1*, Neeraj Kumar1, Sarita Devi1, Gurdev Chand1, Krishan Kumar2 and Satish Kumar3 1Department of Botany and Plant Physiology, 2Pulses Section, Department of Genetics and Plant Breeding, 3Department of Agronomy, CCS HAU, Hisar-125004, Haryana, India *Presenting author: [email protected]
Chickpea (Cicer arietinum L.) is an important food crop grown in India under rainfed condition. The crop usually encounters terminal high temperature (HT) stress during pod filling stage. The objective of the work was to determine the degree of HT effect on two contrast genotypes, viz., ICCV 92944 and HC 5 differing in their tolerance on the basis of net photosynthesis
(Pn), stomatal conductance (gs), SPAD, photochemical efficiency of PS-II measures as chlorophyll fluorescence (CF) and canopy temperature depression (CTD) under field condition. HT stress was given by the changing of two sowing dates, i.e., normal (11th Nov. 2011) and late sown (16th Dec. 2011). Observations were recorded as the temperature rose above 35 ºC after 4, 8, 12 days of exposure. Control readings were taken at the temperature below 30 ºC. With increasing days after exposure to HT a close relationship in decline in photosynthesis, SPAD and CTD values was observed. ICCV 92944 exhibited higher values of Fv/Fm (0.69), SPAD (54.77) but less CTD (-3.56 ºC) as compared to HC-5 having values 0.60, 49.20 and -2.95 ºC, respectively. The variance of these parameters was statistically related to yield stability index. The measured parameters of ICCV 92944 (identified as HT tolerant), declined relatively more slowly than those of HC-5.
Results showed that HT induced stress is detectable from Pn, gs, SPAD, CF, CTD, and these physiological traits can be useful tools to help in screening of chickpea germplasm for HT tolerance.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 177 Session A : Abiotic Stress Adaptation and Management
PA127 (IPPC0967) Evaluation of root system architecture for adaptation to water stress in wheat: deeper rooting Rakesh Pandey1*, Neelu Jain2, Vijay Paul1, G.P. Singh2, R.C. Meena1, Atar Singh1, M.B. Shine2, Vinod Pandey2 and Neha Rai2 1Division of Plant Physiology, 2Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Deep rooting is a useful trait for adaptability under water limited environments. It can be studied in terms of maximum root length and root growth angle. Genotypic variation in root growth in wheat was observed by trench profile wall method after anthesis in thirty three genotypes. Root growth at different depths was expressed as % of total root number density. In the upper profile (0-15 cm) root density was higher in genotypes HD2894, PBW343, GW322, Kukri, HD2824, HI1544 etc. In the middle profile (16-30 cm) the root density was higher in HD2987, DBW17, C306, HD3043, Wyalkechem, HUW468, HD3016 etc. In the lower profile (31-50 cm) the root density was higher in HI1500, Raj3765, HD2687, HUW468, HD2932, HI1531, WH730 etc. These genotypes having higher root density in the lower profile are more adaptable to water stress. However, measuring the root growth in the field conditions is cumbersome and time consuming. Therefore, visualization of root system architecture and measurement of the seminal root angle in the agar gel is a much easier alternative. A narrow root angle between seminal roots is associated with deeper rooting. Variability in the seminal root angle was observed from narrow to wider angles (25oC to 87oC). The relatively drought tolerant genotypes viz. N59, HD2985, DBW- 14, C-306 etc. showed narrow root angles compared to the genotypes for irrigated conditions e.g. HD2967, HD2643, HD2285, HD2824 etc.
PA128 (IPPC0968) Heat stress responses in chickpea Neeraj Kumar1*, Ajit Singh Nandwal1, Krishan Kumar2 and Dharamvir Soni1 1Department of Botany and Plant Physiology, CCS Haryana Agricultural University, Hisar-125004, Haryana, India 2Pulses Section, Department of Genetics and Plant Breeding, CCS Haryana Agricultural University, Hisar-125004, Haryana, India *Presenting author email: [email protected]
High temperature stress (>30ºC) is one of the most evident phenomenon of the global climate change, which limits the plant growth and development in chickpea and leads to drastic reduction in seed yield. With this objective, an experiment was conducted with 15 chickpea genotypes for screening against high temperature tolerance by studying various morpho- physiological traits and seed yield attributes under late (2nd fortnight of December) sown conditions. A significant variation in the quantum yield of PSII (Fv/Fm) was observed among the tested genotypes. The value for Fv/Fm ranged from 0.644 to 0.783 and was highest in ICCV 92944 and lowest in H 09-65. Relative stress injury (RSI %) of leaves varied from 13- 20%, and was considerably high in genotype H 10-22 (20%) and lowest in H10-41 (13%). The canopy temperature depression (CTD) revealed significant differences among genotypes. CTD values ranged between -0.5 (H10-22) and 1.6ºC (H 09-96). Pollen viability was found to be in the range of 65 to 88%, being highest in the genotype ICCV 92944 and lowest in H10-01. Genotypes H09-96, H10-21 and H10-41 gave highest grain yield of 21.7, 19.4 and 18.8 g plant-1, respectively. The genotypes H09-96, H10-21 and H10-41 were identified as heat tolerant lines (>30ºC) based on high values of percentage pollen viability, quantum yield (Fv/Fm), yield attributes and yields but low values of RSI % and maintained cooler canopy temperature.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 178 Session A : Abiotic Stress Adaptation and Management
PA129 (IPPC0969) Brassinosteroids modulate antioxidant defense system under thermal stress in Brassica oleraceae var broccoli Spall Kaur Nirmal K* and Geetika Sirhindi Department of Botany, Punjabi University, Patiala, Punjab-147002, India *Presenting author: [email protected]
Anti-cancerous properties of brocoli are well known, which may be due to the presence of high antioxidants in the form of enzymes and non-enzymes. Brassinosteroids are known for enhancing antioxidant potential of different crops in pico- to micro- molar concentrations. Present study was conducted with the aim to explore the ability of 24-epibrassinosteroid (24-EBL) in enhancing the tolerance level of broccoli under extreme temperature stress. Seedlings of broccoli were primed with distilled water (control) and different concentrations of 24-EBL (10-6, 10-9, 10-12), and grown in seed germinator at 25 0C under controlled conditions of plant growth. To 7 days old seedlings temperature stress was given for three consecutive days and then harvested on 10th day for investigating the content of proline, ascorbic acid along with activity of antioxidant enzymes SOD, POX and CAT. Brassinosteroids increased accumulation of proline by 15% and ascorbic acid by 5%, while activity level of SOD, POX and CAT were enhanced by 18.7%, 34 % and 31%, respectively. The data supports to postulate that 24-EBL exhibited a good potential in mitigating the toxic effect of extreme temperatures on plant growth and enhanced the crop thermo-tolerance without degrading their nutritional and medicinal properties and relatively enhanced the antioxidants of the crop to significant levels thus increasing its anti-cancerous potential.
PA130 (IPPC0983) Salinity induced variations in growth, photosynthetic performance and ionic balance in desi and kabuli genotypes of Cicer arietinum L. (chickpea) under salt stress Ranju Gulati1* and Neera Garg2 1Department of Botany, Dav College, Sector 10, Chandigarh-160036, India 2Department of Botany, Panjab University, Chandigarh-160014, India *Presenting author: [email protected]
Salinity is one of the major environmental stresses affecting crop production especially in arid and semi-arid regions of the world. Chickpea is the most important legume crop for human nutrition globally but at the same time it is highly salt sensitive. However significant variability in terms of salt tolerance has been reported in chickpea germplasm. The present study was carried out to compare salt induced modulations in growth, photosynthetic characteristics and ion accumulation in salt tolerant kabuli (CSG 9651) and salt sensitive desi (DCP 92-3) genotypes of chickpea. The plants of both genotypes were raised under control and NaCl salinity (EC = 4, 6 and 8 dSm-1). The saline treatments were applied15 days after sowing DAS (days after sowing) and continued till harvest. Salinity significantly reduced roots and shoots dry mass and disturbed the root to shoot ratios in both genotypes with significantly higher reductions recorded in DCP 92-3. CSG 9651 possessed higher values for photosynthetic attributes in terms of leaf chlorophyll content with higher Chl a/b ratios and Rubisco activity as compared to DCP92-3. Kabuli genotype CSG 9651 recorded lower sodium uptake, higher potassium and better K/Na ratio which manifested in the form of higher growth and productivity (higher seed yield, HI etc). The study indicated better tolerance ability of kabuli genotype of chickpea subjected to long term salinity when compared with desi.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 179 Session A : Abiotic Stress Adaptation and Management
PA131 (IPPC0986) Introgression of water use and water use efficiency in rice (Oryza sativa L.) and its relevance to drought tolerance N.V. Preethi1*, B.R. Raju, K. Sumanth Kumar, R. Ramachandra and M.S. Sheshshayee Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus, Bangalore-560065, Karnataka, India *Presenting author: [email protected]
Rice is the most important cereal of the world population and is an intensively water consuming crop. It is hypothesized that maintenance of turgor and positive carbon balance is essential for enhancing crop performance under water limited conditions. Global opinion strongly emphasise introgression of these complex traits to achieve improved adaptation to water limited conditions. Towards developing such trait introgressed lines, root donor line (IET15963) was crossed with WUE donor line (Thanu) and a large number of recombinant inbred lines were developed. Among a few trait introgressed lines, KMP175 was chosen for further studies. SSR markers associated with WUE and root traits were used to identify the introgression of the target genomic regions governing the two traits with the integration of water mining and WUE traits. KMP175 displayed superior growth and productivity both under aerobic and water limited conditions. Introgressed line recorded higher photosynthetic rate and stomatal conductance compared with the parents. The field performance of KMP175 was significantly higher than the other rice cultivars recommended for aerobic cultivation. The study provided strong evidence on the importance of introgressing physiological traits to improve water productivity of rice.
PA132 (IPPC1003) Development of activation-tagged mutant population for enhanced water-use-efficiency (WUE) in Oryza sativa ssp. indica Mazahar Moin1*, Achala Bakshi1, A.R. Reddy1, K.V. Rao2, E.A. Siddiqi3, M. Udaya Kumar4 and P.B. Kirti1 1Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India 2Centre for Plant Molecular Biology, Osmania University, Hyderabad, India 3Honorary Professor, Institute of Agricultural Biotechnology, PJTS, Hyderabad, India 4Department of Crop Physiology, GKVK, Bangalore, India *Presenting author: [email protected]
The epitome of any genome research is to identify all the existing genes contained in a genome and investigate their roles. Rice is the most appropriate model crop for generating mutant resource for functional genomics studies because of its economic importance, availability of high quality genome sequence and relatively smaller genome size. In the present study to develop an activation-tagged (gain-of-function) mutant resource for high WUE in indica rice Samba Mahsuri (BPT 5204), we have generated 10,000 Ac/Ds and 3,700 stable enhancer based activation-tagged insertions. These plants are being screened for high WUE by growing under limited water conditions based on morphological (tillering, plant height, stay green character and seed yield), and physiological characterisation (photosynthetic performance through Mini-PAM and carbon isotope composition through IRMS). Two hundres plants have been selected as for high WUE. Their flanking sequence analysis through TAIL-PCR revealed that the insertions are non-uniform and mostly genic, in particular inter-genic (60%). Four phenotypically distinct plants were chosen to study the expression of the candidate genes residing within a 20 kb region, the approximate limit of activation of genes by enhancers from the site of insertion. Expression analysis of 16 genes indicated that activation of a gene is independent from the site of insertion of enhancers, although in most of the cases there was activation of only one of the many tagged genes, however, in one plant two genes were activated. The candidate genes responsible for varied phenotypes are being cloned to further validate their roles in WUE.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 180 Session A : Abiotic Stress Adaptation and Management
PA133 (IPPC1011) Clout of rhizobacterial inoculation on antioxidative enzyme activity in Zea mays under aluminium toxic conditions Priyanka Arora1*, Geeta Singh2 and Archana Tiwari1 1School of Sciences, Noida International University, G.B Nagar, Greater Noida-203201, U.P., India 2Division of Microbiology Indian Agriculture Research Institute, New Delhi-110012, India *Presenting author: [email protected]
This study explores the role of microorganisms in enhancing antioxidative defense mechanism of plants to combat against aluminium stress. In the present study, a pot experiment was conducted to evaluate the effect of bacterial inoculation on antioxidative enzyme activity in maize plants under 60 mM aluminium stress condition. Pots were inoculated with two bacterial isolates belonging to genus Burkholderia and Bacillus. Experimental results revealed that although there was a significant increase in antioxidative enzyme activity under aluminium stressed condition but it was not sufficient enough for complete scavenging of ROS. This activity further enhanced on bacterial inoculation to overcome the imposed aluminium stress. Burkholderial isolate performed better than Bacillus isolate showing an increase of 1.47, 7.1 and 2.17 fold in SOD, CAT and POX activity respectively. Therefore the inoculation of Burkholderial isolate can be suggested for plant growth improvement in maize plant under aluminium toxic conditions.
PA134 (IPPC1016) Ecophysiological adaptation of an extreme halophyte Salvadora persica to high salinity: Implications from ion homeostasis, photosynthesis, PS II photochemistry and antioxidant response Jaykumar Rangani1,2, Asish Kumar Parida1,2*, Ashok Panda1 and Asha Kumari1,2 1Division of Wasteland Research, 2Academy of Scientific and Innovative Research, CSIR-Central Salt and Marine Chemicals Research Institute, Gijubhai Badheka Marg, Bhavnagar-364002, Gujarat, India *Presenting author: [email protected]
Salinity induced modulations in growth, photosynthetic pigments, relative water content, electrolyte leakage, lipid peroxidation, photosynthesis, photosystem II efficiency, and changes in activity of various antioxidative enzymes were studied in the halophyte S. persica treated with various levels of salinity (0, 250, 500, 750 and 1000 mM NaCl). Fresh and dry biomass of shoot and root as well as leaf area decreased at all levels of salinity as compared to control. There was gradual increase in leaf Na+ content with increasing salt concentration up to 750 mM NaCl, but at high salinity(1000 mM NaCl), Na+ content surprisingly decreased and came down to the level observed at 250 mM NaCl. There were non- significant changes in chlorophyll, carotenoid, photosynthetic rate (PN), stomatal conductance (gs) and transpiration rate (E) at low salinity (250 to 500 mM NaCl), however, significant reductions were observed at 750 and 1000 mM NaCl. There were non-significant changes in quantum yield of PSII (ÖPSII), photochemical quenching (qP) and electron transport rate (ETR) at low salinity (250-500 mM NaCl), however, decreased significantly at high salinity (750-1000 mM NaCl). Analysis of isoforms of antioxidative enzymes revealed that the leaves of S. persica have three isoforms each of SOD and POX, two isoforms of APX, one isoform of CAT, and five isoforms of glutathione reductase (GR). There were differential changes in different isoforms of antioxidative enzymes. Our results suggests that the absence of pigment degradation, reduction of water loss and maintenance of water use efficiency, and protection of PSII from salinity induced oxidative damage by coordinated changes in antioxidative enzymes enhanced the high salt tolerance capacity of S. persica.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 181 Session A : Abiotic Stress Adaptation and Management
PA135 (IPPC1024) Expression of the PGPR induced rice OsIAA10 gene improves root growth and confers abiotic tolerance in transgenic Arabidopsis plants Pallavi Agarwal*, P.S. Chauhan, V.A. Sane and C.S. Nautiyal Plant Gene Expression Lab, CSIR-National Botanical Research Institute, Lucknow-226001, India *Presenting author: [email protected]
PGPR are plant growth promoting bacteria, which are known to induce plant growth by variety of mechanisms. Microbial associations within roots and hormonal regulation in the plants have been reported, but the molecular mechanism behind this increase in growth of plants is still unraveled. PGPRs (NBRI isolated strain) help in inducing growth of potted rice plants. Microarray Data of PGPR treated rice plants suggested up-regulation of 494 genes and down-regulation of 1045 genes in PGPR treated rice roots as compared to control rice roots. One of the genes OsIAA10, with 3 folds up-regulation in treated roots, was selected for further studies. OsIAA10 belongs to Aux/IAA family involved in auxin signaling. Phytohormone auxin is known to play a key role in plant development and lateral root formation. Transgenic Arabidopsis lines over-expressing OsIAA10 showed longer roots and better growth under various abiotic stresses.
PA135A (IPPC1039) Elucidation of Glutaredoxin proteins in Dicots. Nilesh Kumar1*, Minesh Jethva1, Saurabh Yadav2 and Hemant R. Kushwaha1 1Synthetic Biology and Biofuels Group, International Centre for Genetic Engineering and Biotechnology, New Delhi-110067, India 2Department of Biotechnology, Hemwati Nandan Bahuguna Garhwal University, Srinagar Garhwal, Uttarakhand-246174, India *Presenting author: [email protected]
Glutaredoxins are small proteins (approx. 11 kDa) containing an active site with a redox-active motif. They regenerate the oxidatively damaged proteins via thiol-disulphide reactions. GRXs acts in anti-oxidant defense by reducing dehydroascorbate, peroxiredoxins, and methionine sulfoxide reductase etc. These proteins are also involved in maintaining the redox homeostasis in many organisms like bacteria, yeast. Apart from their role in antioxidant defense, GRXs are known to bind iron-sulfur clusters (ISC). Depending on the motif present GRXs are classified in two classes: Monothiol which contain ‘CGFS’ and dithiol which contain ‘CxxC’ motif. In the present study, we have characterized GRX family members in various dicots. Fact-finding on GRX in many organisms has revealed a surprising variety of vivid corollaries. In view of that, we have analyzed chromosome localization, protein domain architecture, phylogenetic relationship and subcellular localization of GRX member proteins in various dicots. These analyses have been used to infer the evolution of GRXs in dicots. This study provides cognizance of the domain structure conservation of GRXs in dicots. In analysis provided in the study will assist in developing an understanding of the evolution of redox defense mechanism in plantspecies.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 182 Session B : Signaling and Phototransduction
PB136 (IPPC0114) Exploring signature of natural ageing and CDT by studying AOS and antioxidative enzymes in neem [Azadiracta indica juss] seeds Alok Kumar Sahu* and Subhas Naithani Seed Biology Lab., SoS in Life Sciences, Pt. Ravishankar Shukla University, Raipur-492010 (CG), India *Presenting author: [email protected]
Seed longevity is a resultant of the interaction of seed storage conditions with its genetic and physiological potential. Natural ageing [NA] and control deterioration [CDT] tests are widely used to deduce the pathway of ageing. CDT is rapid test as it accelerates ageing [within days] hence preferred over NA. Natural and CDT protocol revealed conflicting results in regard to ageing. Hence, this investigation is an attempt towards resolving the CDT and NA protocols, mechanism of seed deterioration, and the role of reactive oxygen species (ROS) and their detoxification in the non-orthodox seeds of neem (Azadirachta indica). During NA the 100% viable neem seeds became non-viable in 150 days, whereas under CDT conditions the seeds survived only 24 h. Initially (100% germination), the ROS producing capacity was minimum but increased with prolonged storage, showing comparatively higher levels in the CDT conditions. High levels of SOD, APX and CAT protect the neem seeds from ROS damage thus maintaining high viability. ROS levels accumulated in ageing neem seeds due to impaired antioxidative capacity; decline of intensity of isoenzymes or disappearance of isoenzymes. Appearance of new isoenzymes in the ageing seeds [up to 78% germination] in the NA seeds revealed the basis of intermediate storage behavior of neem seeds. The reduced transcripts of SOD and APX confirmed isoenzymes pattern in NA aged seeds. The pattern of ROS, its antixodative enzymes; quantitative, isoenzymes and gene expression analysis will be discussed to explain the mechanism of NA and CDT in neem seeds in respect to its intermediate storage behavior.
PB137 (IPPC0137) The illusive plant osmosensor: OsHK3b as a possible candidate in rice Ramsong Nongpiur1*, Ashutosh Sharan1, Ratna Karan1, Praveen Soni1, Sneh Singla-Pareek2 and Ashwani Pareek1 1School of Life Science, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Osmotic stress like drought and salinity greatly reduce crop productivity. Rice, one of the major food crops, is extremely sensitive to osmotic stress and it is imperative that we understand the mechanism of how rice responds to osmotic stress. Till date, there is no comprehensive report which shows how plants perceive osmotic stress and thus no bonafide plant osmosensor has been identified. Our quest for identifying an osmosensor has led to the identification of an osmotic stress inducible rice histidine kinase, OsHK3b as a putative candidate. Our results show that OsHK3b has a high degree of structural similarity to EnvZ and SLN1, which are osmosensors in E. coli and yeast, respectively. OsHK3b also shares a high degree of similarity with putative osmosensor of Arabidopsis, AtHK1. Heterologous expression of OsHK3b could not only complement the function of SLN1 in sln1-mutant yeast but also restore osmosensing capabilities to the yeast osmosensing-deficient double mutant, sln1Äsho1Ä, thus showing that OsHK3b can function as an osmosensor in yeast. Overexpression of OsHK3b in rice positively regulates growth as well as tolerance to salinity and drought, while knocking down of OsHK3b expression through RNAi resulted in hypersensitivity to osmotic stress, indicating its positive regulatory role in the osmotic stress response of rice. A probable mechanism of action through the interaction and modulation of OsPhp3 and, subsequently, OsRr22 and OsRr26 is also observed in yeast-two-hybrid and bifluorescence complementation assays.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 183 Session B : Signaling and Phototransduction
PB138 (IPPC0165) Elucidating the two-component mediated signal transduction system in Oryza sativa L. Ashutosh Sharan1*, Praveen Soni1, Sneh L. Singla-Pareek2 and Ashwani Pareek1 1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Our laboratory has been engaged in working out the intricacies of stress physiology and functional genomics of model cereal crop, rice. In this endeavor, we have attempted to dissect the complex interactions amongst an important class of signaling proteins comprising a group designated as two-component system (TCS). The complex rice genome possess large number of TCS genes including 13 histidine kinases (HKs), 5 histidine phosphotransferases (Hpts) and 34 response regulators (Rrs). TCS signaling include specific protein-protein interactions between its various components for signaling. In present study cytosolic fragments of OsHK4 and OsHK5 were used, as full length HKs are membranous protein which can not be used for interaction study using yeast-two hybrid technique (Y2H). We found OsHk4 to interact with OsAhp2, whereas OsHk5 was found to interact with OsAhp1, OsAhp2 and OsPhp1. All Hpts exhibited multiple interactions with different Rrs indicating their function as interaction hubs. We also observed selective interactions between type-A and type-B Rrs. Different pseudo-response regulators showed interactions among themselves. These results constitute strong arguments in favour of a cross talk between signaling mediated by different sensory histidine kinases. We also re- validated these interactions by bimolecular fluorescence complementation assay (BiFC). Interactions of HKs (cytoplasmic fragments) with Hpts were found to be cytoplasmic as well as nuclear where as those of Hpts with Rrs and those among Rrs were found to be localised to the nucleus only. This study provides starting point for further analysis of the molecular mechanisms for the signal transmitted by the TCS into biological processes.
PB139 (IPPC0218) A natural resistant associated macrophage protein from Brassica juncea exhibits a novel interaction with a membrane bound thioredoxin protein Ananya Marik, Haraprasad Naiya, Gairik Mukherjee, Madhumanti Das and Chinmay Saha and Anindita Seal* Department of Biotechnology, University of Calcutta, 35,Ballygunj Circular Road, Kolkata-700019, West Bengal, India *Presenting author:[email protected]
NRAMPs are a family of evolutionarily conserved proton coupled transporters, which transport both essential and non- essential metals in plants. Fifty protein interactors of a Brassica juncea NRAMP, BjNRAMP4.1 were identified through a split ubiquitin yeast two-hybrid screen. Among these interactors, interaction with a membrane bound thioredoxin like protein BjHCF164, homologous to Arabidopsis high chlorophyll fluorescence protein 164 (AtHCF164), was studied in detail. AtHCF164 is implicated in the transfer of reducing equivalents from stroma to luminal proteins during photosynthesis and a knockout is seedling lethal in Arabidopsis. BjNRAMP4.1 and BjHCF164 exhibited dual localization in plants. BjHCF164, together with its reported site of function in chloroplasts, was also found to strongly express in Brassica roots. BjNRAMP4.1 localized in chloroplasts and epidermal membranes in leaves and membrane in roots. In keeping with such distinct tissue-dependent localization, the proteins interacted in chloroplasts in leaves and membrane in roots. The interaction was found to increase in the presence of excess manganese both in yeast as well as in plant roots. An interaction network created from Arabidopsis homologs of the BjNRAMP4.1 interactors was highly enriched in proteins central to chloroplastic/cellular ROS signaling. Homology modeling and docking studies suggested that BjNRAMP4.1 could be potentially redox regulated by BjHCF164, while BjHCF164 could be regulated through hydrophobic interaction through residues within the thioredoxin domain. Our results suggest that interaction between BjNRAMP4.1 and BjHCF164 may be a potential link between metal homeostasis and chloroplastic/cellular redox function.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 184 Session B : Signaling and Phototransduction
PB140 (IPPC0296) Regulation of microRNA biogenesis by mitogen activated protein kinases in plants Prakash Kumar Bhagat* and Alok Krishna Sinha National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
MicroRNAs (miRNA) are 21 -24 nucleotide in length transcribed by RNA polymerase II from mir gene locus. The processing of primary miRNA (pri-miRNA) transcript to precursor miRNA (pre-miRNA) followed by generation of mature miRNA/ miRNA* duplex is carried out by a microprocessor/biogenesis complex. The microprocessor complex comprises of RNaseIII endonuclease, DCL1 (DICER –LIKE1 protein), DRB1/ HYL1 (double stranded RNA binding protein /HYPONASTY LEAVES1), SE (zinc finger protein SERRATE) and TGH (G- patch domain protein TOUGH). The mature miRNA/miRNA* duplex is transported to the cytoplasm where it is processed further, finally targeting mRNA trancript to regulate its expression. Earlier studies have suggested a role of dephosphorylarion of DRB by a phosphatase, CPL1 is required for the biogenesis of miRNA. While another report from our lab suggested that DRB1 is phosphorylated by MPK3 and that it negatively regulates miRNA biogenesis. We here report characterizing of OsDRB1 in rice using surface plasmon resonance (SPR) technology. Deletion construct of DRBs have been prepared to map the MAPK phospho site/s on these proteins. To get an insight of these interactions and to elucidate the biological function of DRBs and OsMPK3, a transient knockdown protocol using VIGS vector has been standardized. In nut-shell the regulation of DRBs during miRNA biogenesis by MAPK will be discussed.
PB141 (IPPC0300) Investigation of novel regulators involved in MAP kinase and light signalling pathways during Arabidopsis seedling development Alok Krishna Sinha and Deepanjali Verma* Plant Signaling, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Light modulates the changes in the gene expression that occur at various developmental stages of plants in defined tissues. These changes take place mainly at the transcriptional level and lead to the regulation of transcription of specific genes that contributes light regulatory developments. MYC2, a basic helix loop helix transcription factor is known as a positive regulator of jasmonic acid mediated inhibition of root growth. In an earlier study it has been reported that a MAP kinase module plays a key role in MYC2 mediated light signalling. The study demonstrates that MPK6 is activated by MKK3 in blue light (BL) and that this process is MYC2 dependent. It is also shown that MPK6 physically interacts with and phosphorylates MYC2 which binds to the promoter of MPK6 and represses its expression. In the present work we have identified a phosphatase, MKP1 as an interacting partner of MPK6 and regulator of MPK6-MYC2 module. The interaction was confirmed by in-vitro phosphorylation assay, which showed that MKP1 dephosphorylates MPK6 in the presence of blue light thus regulating MPK6-MYC2 module, which is a very crucial step and needs to be tightly controlled. To identify the upstream MAP3K which activates MKK3 in this pathway, we have selected 6 MAP3Ks showing upregulation in BL on the basis of in-silico microarray analysis. To further substantiate we performed gene expression analysis which identified 3 MAP3Ks that might be involved in the pathway. We intend to identify the specific MAP3K involved in the activation of MKK3-MPK6-MYC2 module mediated light signalling.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 185 Session B : Signaling and Phototransduction
PB142 (IPPC0353) Functional analysis of Arabidopsis FCS-LIKE ZINC FINGER gene family suggests their role in the regulation of plant growth and stress mitigation Muhammed Jamsheer K., Dhriti Singh*, Chanchal Thomas Mannully, Yadvendra Singh Chattha and Ashverya Laxmi National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
The FCS-LIKE ZINC FINGER (FLZ) gene family is characterized by the presence of a solitary FCS-LIKE ZINC FINGER (FLZ) domain which is involved in mediating protein: protein interaction. Many members of this gene family are implicated in sugar signaling, biotic and abiotic stress response. In this study, we identified that the Arabidopsis thaliana FLZ proteins interacts with a wide variety of proteins which are involved in vegetative and reproductive development, developmental phase transition, stress response, etc. In some cases, even the evolutionary distant proteins were found to share common interacting proteins suggesting a high degree of functional redundancy in this gene family. Besides, these zinc fingers were found to be interacting with other members of this protein family and in many cases this interaction was found to be important for maintaining proper sub-cellular localization. The FLZ domain was found to be the canonical interacting module for interacting with other proteins while the N-terminal region was found to important for interacting with the proteins of the same protein family. The knock-out mutants of these genes were found to have an impact on normal growth and adaptive growth under osmotic stress suggesting the role of this gene family in plant growth and development and adaptive growth under stressed conditions. Further molecular and physiological analysis need to be done to decipher the exact molecular functions of these genes.
PB143 (IPPC0367) Role of stress-induced F-Box proteins in ABA and reactive oxygen species mediated signalling in Arabidopsis and rice Avantika Bhaskar1, Eshan Sharma1* and J.P. Khurana2 1Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi 110021, India 2Department of Plant Molecular Biology, Interdisciplinary Centre for Plant Genomics and University of Delhi, South Campus, New Delhi-110021, India *Presenting author: [email protected]
Protein degradation is an important aspect of plant growth and development mediated by the ubiquitin/26S proteasome pathway, allowing cells to rapidly respond to changes in environment. Here, we present functional characterisation of an F-box protein encoding gene OsFBX in rice and its orthologous genes in Arabidopsis. Quantitative RT-PCR expression analysis revealed induction of OsFBX under various abiotic stresses with high transcript levels specifically under drought in Oryza sativa cv IR64. Our analysis also showed high levels of OsFBX during seed development followed by panicle developmental stages. Transiently expressed OsFBX (fused with reporter gene encoding GFP) localized to the nucleus indicating it as a part of proteasome complex that is abundant in the nucleus. Ectopic expression of OsFBX in Arabidopsis thaliana led to a hypersensitive phenotype in response to ABA, glucose and osmotic stress. The corresponding orthologous mutants in Arabidopsis showed hyposensitivity to ABA mediated responses such as inhibition of seed germination, root and seedling growth in presence of ABA, NaCl and mannitol. Further, short duration ABA treatment increased ROS levels in Arabidopsis lines overexpressing OsFBX compared to reduced levels in Arabidopsis mutants. The ABA sensitivity of mutants and transgenic plants also correlated with faster water loss in mutants. Thus, the F-box protein analysed in this study may positively regulate ABA-mediated plant responses under abiotic stress conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 186 Session B : Signaling and Phototransduction
PB144 (IPPC0378) Effects of oxidative and photooxidative stress on reactive oxygen species and proline metabolism in leaf discs of Pisum sativum A.S. Raghavendra and Vetcha Aswani* Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India *Presenting author: [email protected]
Plants are continuously exposed to biotic and abiotic stresses. On exposure to stress, the reactive oxygen species (ROS) accumulate and these ROS should be scavenged. We have examined the patterns of ROS accumulation and the consequences in leaf discs of pea, Pisum sativum. We employed oxidants, which induce ROS in different cellular compartments: acifluorfen methyl ester (AFM) and paraquat (PQ) (both in chloroplasts), menadione (MD) (mitochondria) and abscisic acid (ABA) (plasma membrane). The accumulation of H2O2 and superoxide were estimated by the oxidation and reduction of DAB and NBT, respectively. The ROS levels were higher in AFM and PQ treated samples compared to that of MD and ABA. Among the four antioxidant enzyme activities studied, the increase in the activity of superoxide dismutase (SOD) and glutathione reductase (GR) were higher than that in ascorbate peroxidase (APX) and catalase (CAT). Proline, an osmolyte, increased significantly under stress. Among key enzymes of proline metabolism, slight increase in the activity of pyrroline 5 carboxylate synthase (P5CS) compared to the marked decrease in proline dehydrogenase (ProDH) indicated that increased proline levels were due to the restricted proline oxidation. Finally, the change observed in the antioxidant metabolism was validated by studying physiological response by oxygen electrode. These results indicated that the leaf photosynthesis was decreased in all treatments; maximum was with PQ, whereas, respiration was lowest in MD treatment. The protein levels and gene expression pattern of the antioxidant enzymes (CAT, APX, GR) and of proline metabolism (P5CS and ProDH) are also studied.
PB145 (IPPC0385) Signaling components and their interaction during stomatal closure by microbial elicitors, in comparison with the effects of abscisic acid Gunja Gayatri1*, Appa Rao Podile1, Mallikarjuna Rao Puli1, Srinivas Agurla1, Kazuyuki Kuchitsu2 and A.S. Raghavendra1 1Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-50046, India 2Department of Applied Biological Science, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba *Presenting author: [email protected] Plant stomata act as the main access points for the entry of the pathogens. Stomatal closure restricts the entry of the pathogens, becoming one of the plant innate immune responses. Such closure is mediated by microbial elicitors, produced from either microbes or plant tissue. We therefore studied effect of three elicitos: flg22 (a 22 amino acid peptide from eubacterial flagellin), harpin (from Pseudomonas syringae pv. syringae), cryptogein (from Phytophthora cryptogea) on stomata of Arabidopsis thaliana. All the three elicitors (flg22, cryptogein and harpin) induced marked stomatal closure. The stomatal closure, for e.g. by ABA, involves several signaling components, including reactive oxygen species (ROS), nitric oxide (NO), G-proteins, calcium and protein kinases/protein phosphatases. We therefore examined the patterns of ROS and NO in guard cells. The elicitors raised markedly the levels of ROS or NO in guard cells. Time course monitoring of ROS or NO, and studies using ROS or NO modulators on stomatal closure revealed that ROS acted upstream of the NO during stomatal signaling by these elicitors, similar to ABA. Participation of signaling components was further confirmed by mutants. Impaired stomatal closure and decreased ROS or NO levels by these elicitors in guard cells of atrbohD/F confirmed that ROS was essential during stomatal closure. The partial reversal of stomatal closure in nia1 and nia2, decreased NO production indicated the essentiality of NO production during elicitor triggered stomatal closure. A scheme of signaling events during stomatal closure by microbial elicitors would be proposed.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 187 Session B : Signaling and Phototransduction
PB146 (IPPC0423) Abiotic stresses, tissue specificity and diurnal rhythmicity act as multiple regulatory switches to control transcription of members of two-component system in rice Praveen Soni1#*, Priyanka Gupta1, Sneh L. Singla-Pareek2 and Ashwani Pareek1 1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India #Present Address: Department of Botany, University of Rajasthan, Jaipur-302004, Rajasthan, India *Presenting author: [email protected] Two-component system is his-to-asp phosphorelay based signaling machinery that mediates perception, signal transduction and response to environmental stresses. Though, TCS members have already been reported in rice but their functions are yet to be identified. To gain insight into the role of TCS members, their expression responses to environmental stresses and ABA hormone were analyzed in contrasting genotypes-IR64 and Pokkali at seedling stage. We observed their differential regulation in genotype-dependent and stress-specific manner. Transcript profiling distinguished OsHk3, OsPhp3, OsRr21 and pseudo-response regulators as ABA inducible and multiple stress-responsive TCS members. We also carried out their transcript analysis in different plant tissues and different time points of a day. This analysis revealed that TCS machinery is regulated by not only various abiotic stresses, but also by the tissue specificity and diurnal rhythmicity. Some members showed prominent expression in all tissues while some others such as Authentic histidine phosphotransferases, OsRr21 and pseudo-response regulators exhibited high expression in mature panicle and green parts. Our data suggests that TCS members might play important role in organ development during juvenile stage and panicle development or grain filling at mature stage. Expression of most of the TCS members showed robust oscillation with peaks at different time points of a 12h light/12h dark cycle. Thus, we report an intricate and dynamically intertwined regulatory network of TCS members, as influenced by the abiotic stress, ABA, tissue and diurnal rhythm. This study provides important clues for thorough investigationof diverse roles of the various TCS family members in rice.
PB147 (IPPC0438) Polyamines induce stomatal closure and increase reactive oxygen species and nitric oxide levels in Arabidopsis thaliana Agurla Srinivas*, Gunja Gayatri, A.S. Raghavendra Department of Plant Sciences, University of Hyderabad, Hyderabad-500046, India *Presenting author: [email protected]
Stomata are tiny pores present mostly on leaves and regulate the gas exchange as well as transpirational loss of H2O into the atmosphere. The opening and closing of stomata are due to changes in turgidity of guard cells. Several abiotic and biotic factors induce stomatal closure. For example abscisic acid (ABA), a drought inducible phytohormone, induces stomatal closure in different plant species. Signalling components like reactive oxygen species (ROS), reactive nitrogen species (NO), Ca2+, cytosolic pH, sphingolipids, phospholipids and MAP kinases were all involved in ABA induced stomatal closure. Polyamines (PAs) are ubiquitous polycationic nitrogenous compounds, that are associated with plant adaptation to abiotic factors. We have studied the effect of three polyamines: putrescine (Put), spermidine (Spd) and spermine (Spm), on stomatal movement in Arabidopsis thaliana. Among these, Put was more effective than Spd and Spm in inducing stomatal closure. The levels of ROS and NO in guard cells on exposure to the polyamines were monitored by using fluorescent probes like CM-H2DCFDA and DAF-2DA respectively. The levels of ROS and NO in guard cells were increased upon treatment with the PAs. Thus, the patterns of ROS and NO levels in guard cells, corresponded with stomatal closure. The restriction of stomatal closure by 2-Phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO, NO scavenger) and N-nitro-L-Arg-methyl ester (L-NAME, a nitric oxide synthase inhibitor) confirmed the role of NO during polyamines induced stomatal closure. The participation of NADPH oxidase, polyamine oxidase and peroxidases in the ROS production and stomatal closure is being examined.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 188 Session B : Signaling and Phototransduction
PB148 (IPPC0444) Characterization of a potato class-I KNOX gene- potato homeobox 15 Ameya Mahajan1, Kirtikumar Kondhare1*, Mohit Rajabhoj1, Tejashree Ghate1, Nevedha Ravindran1, Amit Kumar1, Farhat Habib1, Sundaresha Siddappa2 and Anjan Banerjee1 1Indian Institute of Science Education and Research, Pune-411008, Maharashtra, India 2Division of Plant Protection (Plant Physiology), Central Potato Research Institute, Shimla-171001, India *Presenting author: [email protected]
Potato Homeobox 15 (POTH15) is orthologous to Shoot Meristemless (STM) in Arabidopsis. POTH15 belongs to potato KNOX-I family gene. KNOX genes have been characterised in several species, but very few studies are available in potato. The major objective of this study is to characterise POTH15 and identify its targets involved in potato development. In the first experiment, we investigated whether POTH15 transcript shows differential expression under tuber-inducing short day v/s tuber non-inducing long day conditions in a photoperiod sensitive potato cultivar. qRT-PCR analysis was conducted in a tissue-specific manner and it was observed that shoot tips and stolons showed high levels of POTH15 mRNA under short day conditions. In the second experiment, we studied the expression of POTH15 promoter. POTH15 promoter showed a wide expression pattern (e.g. apical and axillary meristems, petiole, stolon tip, tuber eyes, tuber pith) suggesting that POTH15 has a diverse role in potato development. In subsequent experiments, we have undertaken a constitutive over-expression approach to identify the functions and targets of POTH15. In POTH15 over-expression lines, number of morphological traits, such as leaf, stem, tuber development, were altered. POTH15 over-expression lines also tuberised earlier than wild-type plants under in vitro conditions. We further performed a comparative RNA-seq analysis with wild-type and POTH15 over-expression lines, which identified 435 differentially expressed genes involved in key biological processes. Selective candidate genes were validated by qRT-PCR for their differential expression in wild-type and POTH15 over-expression lines. To summarise, this study suggests that POTH15 controls diverse developmental processes in potato.
PB149 (IPPC0452) Functional characterization of OsSnRK2 gene in rice Sanya Shrivastava*, Santosh Kumar Vinjamuri, Shashank Kumar Yadav, Pooja Dhuppar, Viswanathan Chinnusamy Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Sucrose non-fermenting-1 related protein kinase (SnRK) is a unique family of serine/threonine protein kinases associated with energy and abiotic stress signalling in plants. They are divided into three subfamilies SnRK1, SnRK2, and SnRK3. In rice, 10 members SnRK2 family were identified and shown to be activated by salt stress. However, their role in abiotic stress tolerance is poorly understood. Microarray data analysis of OsSnRK2.4 revealed that it is up-regulated mainly by dehydration and salinity treatments. In this study, rice OsSnRK2.4 gene was cloned and characterized. OsSnRK2.4 gene was cloned in pCAMBIA1300 vector under stress inducible AtRD29A promoter and constitutive ZmUBI promoter. Transgenic rice plants were developed using these constructs and confirmed through PCR, qRT-PCR and southern blotting. Post- germination analysis of OsSnRK2.4 overexpressing transgenic plants showed better shoot length and root length as compared with WT plants. Analysis of drought tolerance of RD29A:OsSnRK2 transgenic rice lines in pot culture under greenhouse conditions revealed that RD29A:SnRK2 transgenic rice lines are more tolerant than non-transgenic lines in terms of maintenance of membrane stability index, RWC, survival and recovery growth. Further studies on detailed characterization of SnRK2 will elucidate their role in development and abiotic stress responses of rice.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 189 Session B : Signaling and Phototransduction
PB150 (IPPC0490) Transcriptional regulation of BREVIS RADIX (BRX) in progenitors of bread wheat with contrasting root traits Pramod Awakale, Sneha Tiwari and Monika Dalal* ICAR-National Research Centre on Plant Biotechnology, New Delhi-110012, India *Presenting author: [email protected]
Deep root system is an important drought avoidance mechanism in plants. The highly plastic nature and difficulty in phenotyping deters the use of root traits in breeding programs. Identification of genes/alleles and markers associated with RSA can expedite the use of this trait to improve yield of crop plants. The diploid progenitor species of bread wheat (T. aestivum L.) namely T. monococcum (A genome), Aegilops tauschii (D genome) and Ae. speltoides (S genome, closest to B genome), are potential resources for various agronomic traits including abiotic stress tolerance. However, these species have not been explored for inductive root traits. Therefore to characterize this untapped genetic resource, seedlings of nine accessions of diploid wheat (three accessions each representing A, B and D genomes) were phenotyped for inductive root growth under two levels of osmotic stress viz. -0.5 (S1) and -1.48 bars (S2) for 10 days. Significant genetic variability was observed for root traits among the accessions with different genomes. The BREVIS RADIX (BRX) transcription factor controls cell proliferation and elongation in growth zone of root tip and explained 80% of variance in root length in Arabidopsis. Hence, BRX expression was analysed under control and stress conditions in the diploid accessions with contrasting root traits. The analysis of BRX expression and allelic variation will help in understanding the evolutionary role of BRX in root development and growth in wheat.
PB151 (IPPC0493) Interaction specificity of heterotrimeric G-protein beta (Gâ) and gamma (Gã) subunits in Brassica juncea Gulab Arya*, Roshan Kumar and Naveen Bisht National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Heterotrimeric G-proteins (G-proteins) are guanine nucleotide (GTP and GDP) binding proteins that constitute Gα, Gβ and Gγ subunits, and regulate many aspects of physiological development and cell signaling across phylas. Various loss/ gain of functional studies in model plants Arabidopsis and rice revealed that G-protein βγ dimers are involved in many biological processes like hormone signaling, stomatal opening and closing, resistance against necrotrophic fungi, controlling shape and size of leaf and seeds, and root development. However, our understanding about G-protein signaling complex from any polyploid crop is limited. In current study we identified multiple homologs of G-protein β and γ subunits from allotetraploid B. juncea (AABB), an economically important oilseed crop of Brassica genus. Protein-protein interaction between different G-protein β and γ subunits using yeast based assays showed a high degree of differential interaction specificity among the multiple G-protein subunits. Amino acid sequence alignment of multiple G-proteins revealed presence of divergent key residues across Gγ homologs. Site directed mutagenesis (SDM) followed by yeast based interaction assays could delineate key residues imparting interaction specificity across B. juncea G-protein β and γ subunits.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 190 Session B : Signaling and Phototransduction
PB152 (IPPC0499) Arabidopsis G-protein coupled receptor (GCR1) regulates cold, heat and salt stresses Navjyoti Chakraborty*, Navneet Singh, Nandula Raghuram and Kanwaljeet Kaur University School of Biotechnology, Guru Gobind Singh Indraprastha University, Sector 16C, Dwarka, New Delhi-110075, India *Presenting author: [email protected]
G-proteins and G-protein coupled receptors (GPCRs) have been implicated in regulating the plant’s response to some or the other biotic/abiotic stress, but a comprehensive assessment of the role of GPCR in stress response has not been done so far, partly due to the recent controversy regarding the existence and need for in plant G-protein signalling. We isolated a GCR1-knock-out mutant (gcr1-5) in Arabidopsis thaliana and performed its transcriptome analysis using microarrays in comparison with the wild type to understand the genome-wide roles of GCR1. Using a stringent cut off of log2FC 1.0 and p-value d”0.05, we found 350 differentially expressed genes (DEGs) in the mutant. More than 100 of them were found to be involved in response to biotic/abiotic stress. Further analyses revealed that out of these, 57 DEGs (49 up/8 down) belong to abiotic stress response category. We validated the effects of cold, heat and salt stresses on the GCR1 mutant and WT using standard and biochemical parameters. We also validated some of the stress-responsive DEGs by qRT-PCR. Overall, we found that gcr1-5 is most tolerant to cold, followed by heat and salt, in terms of all the above parameters. These results clearly show the important role of GCR1 in mediating the plant’s response to multiple abiotic stresses. Considering that G-proteins are also implicated in many of them, our data also indicate the possibility that GCR1 may act as a GPCR in G-protein signalling in stress response.
PB153 (IPPC0558) Cloning and expression analysis of ABA receptor(s) in rice Shashank Yadav1, Pooja Dhuppar1*, Santosh Vinjamuri1, Sanya Shrivastava1, Pragya Yadav1, Rakesh Verma1, Bhupendra Chaudhary2 and Viswanathan Chinnusamy1 1Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India, 2School of Biotechnology, Gautam Buddha University, Greater Noida, Uttar Pradesh, India *Presenting author: [email protected]
Abscisic acid (ABA) is an essential phytohormone which is involved in a host of biological processes, including plant development and responses to biotic and abiotic stresses. The hunt for ABA receptors culminated in the identification and characterization of PYRABACTIN RESISTANCE1 (PYR1)/PYR1-like (PYL)/ Regulatory Component of ABA Receptor (RCAR) proteins as ABA receptors (ABARs) in Arabidopsis. ABA-bound ABAR inhibits phosphatase type 2C proteins. This results in relieving of SnRK2 inhibition by PP2C, and then SnRN2 mediated activation of effector ion channels and transcription factors. The specific functions of individual members of ABARs in developmental and stress responses are being currently investigated worldwide. This study was conducted to identify and characterize of ABAR family genes from rice. Phylogenetic analysis grouped these ABARs in to three distinct subfamilies: the monomeric receptors formed subfamily I and sub-family II, while the dimeric receptors formed subfamily III. Only the subfamily I genes contain introns. Expression analysis of ABARs in different tissues and stress conditions revealed that differential expression patterns. This suggests non-redundant roles of ABARs in development and stress responses. All the ABA receptors were cloned from Nagina22 in pCAMBIA1300 under the control of stress inducible promoter AtRD29A for Agrobacterium mediated genetic transformation of rice. Comparison of abiotic stress tolerance of rice transgenics overexpressing ABAR9/ABAR10/ABAR11 with non-transgenic plants suggests that ABARs are potential candidate genes for improving drought tolerance of rice.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 191 Session B : Signaling and Phototransduction
PB154 (IPPC0576) Functional characterization of Med14 in Arabidopsis Sourobh Maji* and Jitendra Thakur National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Mediator complex is an important transcriptional co-regulator and consist of 31 subunits in Arabidopsis. AtMed14 is a part of tail modular subunit of plant Mediator complex. It is conserve in eukaryotes and contains N-terminal MED14 domain, which is conserve throughout eukaryotic kingdoms. In metazoans it is known to be involved in androgen signaling, intracellular steroid hormone receptor signaling, stem cell maintenance and positive regulation of RNA polymerase II transcriptions. Recent studies in plants revealed its important role in growth, development and survivability. Mutation in Med14 has dramatic alteration in cell proliferation, cold response, salicylic acid induced defense response and SAR response. We found that a homozygous T-DNA insertion line of Med14 was embryo lethal in which embryo got arrested at different stages of embryo development. Most of the seed obtained from heterozygous lines were defective in seed germination. These defects in the seed germination were due to defect in the secretion of mucilage from columella cell of seed coat. This particular defect in seed mucilage secretion phenotype is identical to leunig_homolog (luh) and mucilage modified 2 (mum2) mutants. MED14 physically interacting with LUH and regulate the expression of MUM2, an enzyme needed for mucilage extrusion. In addition, MED14 also physically interacts with well known auxin regulator LEUNIG (LUG) and AINEGUMENTA (ANT), suggesting its role in transcriptional regulation during auxin signaling. Thus, our study reveals importance of Med14 in auxin-mediated processes in early stage of plant development.
PB155 (IPPC0675) Functional characterization of a central effector of light signaling, AtHY5, ortholog in rice Naini Burman*, Akanksha Bhatnagar and Jitendra P. Khurana Interdisciplinary Centre for Plant Genomics and Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India *Presenting author: [email protected]
Light plays a very important role in plant’s life as they are sessile and photoautotrophic. Plants have developed an intricate network of light signaling components, which interact with almost all the important pathways. This light signaling cascade is highly hierarchical with photoreceptors like phytochromes and cryptochromes being at the top. They are followed by early signaling factors like HFR1 and central integrators like COP1. Then lies the downstream effectors like HY5. Most of the components of light signaling pathway have been characterized in Arabidopsis, a model dicot plant. One of the orthologs of AtHY5 in rice, encoded by OsbZIP48, has been functionally characterized using transgenic approach. Over-expression transgenics of OsbZIP48 caused dwarf phenotype and accumulation of more chlorophyll content. There was a high degree of floret sterility, which severely affected the grain yield. RNAi transgenics of OsbZIP48 segregated into two types of seedlings in T2 generation; one which resembled wild type and vector control in morpology and the other which showed lethal phenotype. Microarray analysis revealed that a number of hormonal pathways like gibberellin synthesis pathway were affected in these transgenics. These data obtained by analysis of rice transgenics for OsbZIP48 provide evidence that rice ortholog of AtHY5 performs overlapping functions with HY5 as well as it plays some monocot specific roles in regulating plant development.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 192 Session B : Signaling and Phototransduction
PB156 (IPPC0689) OsTLP, an F-box containing TUBBY protein in rice, regulates stress signaling and development Nitin Jain*, Satyam Vergish and Jitendra P. Khurana Department of Plant Molecular Biology, Interdisciplinary Centre for Plant Genomics, Delhi University, South Campus, New Delhi-110021, India *Presenting author: [email protected]
The TUBBY genes were identified in mammals as a small group of 4-5 members based on a mutation in mice that caused obesity on maturation. In animals, they also regulate insulin pathway, retinal maintenance, neuronal development, etc. These class of proteins, harboring a C-terminal TUBBY domain, are bi-partite transcription factors that translocate to the nucleus from the plasma membrane on perceiving certain signals and have both DNA binding and transcriptional activation properties. In plants, these C-terminal TUBBY domain containing proteins are invariably coupled with a conserved N- terminal F-box domain and are thus called as TLPs (TUBBY LIKE PROTEINS). Both Arabidopsis and rice have nearly 14 TLPs that regulate diverse functions such as abiotic stress signaling, hormone signaling, plant-pathogen interactions and so on. Our lab identified a few F-box proteins in rice that are regulated under different light conditions including an OsTLP. This OsTLP has been further characterized and found to be upregulated under blue light and is also under circadian control as its expression dips at midnight. The gene also showed upregulation under salt and drought stress and in ovary tissues. The protein harbors a canonical F-box domain which interacted with OsSKPs showing that it is a component of a functional SCF-type E3 ligase. The protein was found to be localized to the plasma membrane that shifted to the nucleus under stress conditions. The over-expression lines in Arabidopsis showed better over-all growth and involvement of the gene in various abiotic stresses. The data thus far indicate involvement of the OsTLP in abiotic stress signalling and contribution towards overall growth and development.
PB157 (IPPC0690) Functional characterization of an OsbZIP gene in rice for its possible role in light signaling Akanksha Bhatnagar*, Naini Burman and Jitendra P. Khurana Interdisciplinary Centre for Plant Genomics & Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India *Presenting author: [email protected]
Plant growth, development and response to environmental stimuli are determined by programmed expression of its genes which are regulated by transcription factors. These transcription factors are usually members of multigene families. One of the largest and most diverse transcription factor families in eukaryotes is the basic leucine zipper (bZIP) transcription factor family. Over 120 bZIP transcription factor genes have been identified in different plants, out of which 89 have been identified from rice and 67 from Arabidopsis. Phylogenetic analysis of the bZIP transcription factors across plant species has predicted a majority of the rice bZIP transcription factors to be orthologs of Arabidopsis bZIP transcription factors. One of the most well characterized bZIP transcription factor in Arabidopsis, HY5, has been predicted to have three orthologs in rice, one of which (OsbZIP48) has already been charcaterized in our laboratory for its orthologous function to AtHY5. In this presentation, the function of another member of this sub-group has been presented. This OsbZIP gene too has been found to functionally complement the hy5 mutant in Arabidopsis. Expression levels of this ortholog of HY5 have been checked in different developmental stages as well as in different tissues of rice by real time PCR. Analysis of its expression in 2-10 day old light as well as dark grown seedlings has also been done and these results indicate difference in expression patterns of both these orthologs in rice. Transgenic analysis in both rice as well as Arabidopsis, however, points towards their functional similarity.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 193 Session B : Signaling and Phototransduction
PB158 (IPPC0707) Promoter analysis of GA2ox1, a gibberellin catabolism gene, reveals its role in blue light-mediated photomorphogenesis in Arabidopsis Sushma Mishra* and Jitendra P. Khurana Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India *Presenting author: [email protected]
The young light-grown seedlings characteristically display short hypocotyl, open apical hook with expanded cotyledons and plastid development in dicot systems like Arabidopsis. This photomorphogenic development occurs by the co- ordination of light signals perceived by photoreceptors and endogenous cues like phytohormones. One such example is the interaction between cryptochromes (CRYs), the blue light-sensing photoreceptors, and the phytohormone gibberellin (GA). Prompted by the observations made in our laboratory on Brassica CRY1-ox transgenics, which showed reduced plant height accompanied by altered transcript levels of GA metabolism genes, it was decided to work out the intricacies associated with this cross-talk in Arabidopsis. The real-time PCR analyses confirmed that blue light stimulates the expression of AtGA2ox1, a gene encoding GA-catabolism enzyme. Moreover, Arabidopsis transgenics containing GA2ox1 promoter-reporter (GUS) construct showed higher promoter activity in blue light grown seedlings in comparison to the dark grown ones; it is likely that higher activity of GA catabolism enzyme leads to low availability of bioactive GA levels and hence causing suppression of elongation, resulting eventually in shorter plant height. The trans-acting factors interacting with GA2ox1 promoter were identified by yeast-one-hybrid assay, and the interaction confirmed by electrophoretic mobility shift assay. Work is in progress to elucidate the precise role of these factors in light regulated expression of GA2ox1.
PB159 (IPPC0853) Nitric oxide in plants: the biosynthesis and cell signalling functions of a fascinating molecule in plants Luis Mur, R. Ratcliffe2, Werner Kaiser3 and Kapuganti Jagadis4* 1Institute of Biological, Environmental and Rural Sciences, University of Aberswyth, Wales, UK 2Department of Plant Sciences, University of Oxford, Oxford, UK 3Department of Botany, University of Wuerzburg, Wuerzburg, Germany 4National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected] The free radical nitric oxide (NO) emerged as an important signal molecule in plants. Various reductive, oxidative pathways operative for NO biosynthesis. The reductive pathway utilizes nitrite as substrate, which is exclusively produced by cytosolic nitrate reductase (NR) and mitochondria. Using plant mitochondria, we show that nitrite reduction to NO is strongly increased with the decrease in oxygen, which is a general consequence under flooding stress. The reaction is linked to ATP synthesis under hypoxia. NO diffuses from the mitochondria to the cytosol where it is scavenged to nitrate by the non-symbiotic haemoglobins. I will present experimental data supporting these reactions and showing that under low oxygen, the plant mitochondrion serves as a nitrite: NO reductase and becomes a major component in the anoxic nitrogen cycling where it directly contributes to a decrease of cell reduction level and to a limited ATP synthesis. We also found that NO inhibits aconitase and increases in citrate levels which then act as a potent inducer of AOX pathway. The NO production, inhibition of aconitase, and induction of AOX leads to a shift of plant metabolism towards amino acid biosynthesis. Under normoxic conditions NO regulates respiration, internal oxygen, carbohydrate utilization, and ROS levels in roots. We show that a decrease in NO, leads to a drop in internal oxygen, an increase in glucose consumption, further elevation of ROS. Thus, NO is required for maintaining steady-state oxygen concentrations and to keep ROS low. Under pathogen attack nitric oxide is important signal for plant resistance and development of hypersensitive response and NO also play role in suppression of high concentration of NO induced by root pathogens.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 194 Session B : Signaling and Phototransduction
PB160 (IPPC0854) Nitric oxide plays a role in the establishment of aerenchyma formation in wheat (Triticum aestivum L.) Aakanksha Wany*, Sonal Mishra, Rythm Vanvari and Jagadis Kapuganti National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Plant have several adaptive strategies for survival under hypoxic conditions. One of the adaptive strategy is aerenchyma formation in roots. Lysigenous aerenchyma contributes hypoxic tolerance in roots, by providing an internal aeration system for the transfer of oxygen from the shoot to root. However, aerenchyma formation requires the death of cells in the root cortex. Nitric oxide is known to induce cell death in plants during incompatible plant pathogen interactions, therefore there is a possibility that NO could play role in lysigenous cell death during aerenchyma formation. In order to understand whether NO play role in aerenchyma formation, we subjected nitrate grown wheat (HD 3086 and HD 2967) plants to anoxia for 24, 48 and 72 hours and then compared aerenchyma formation and nitric oxide formation with normoxic conditions. We found that under anoxic condition NO production has increased which was visualised by DAF-FM fluorescence. Under anoxic treatment roots have increased levels of aerenchyma formation. Application of nitric oxide scavenger cPTIO reduced the levels of aerenchyma formation. Metabolic, gene expression studies revealed that NO plays an important role in establishment of aerenchyma thus contribute for flooding survival.
PB161 (IPPC0862) Hormone profiling reveals the influence of He-Ne laser on Brinjal (Solanum Melongena L.) var. Mattu Gulla P.S. Swathy1*, K.R. Kiran1, K.K. Mahato2, K. Satyamoorthy3 and A. Muthusamy1 1Department of Plant Sciences, 2Department of Biophysics, 3Department of Cellular and Molecular Biology, School of Life Sciences, Manipal University, Planetarium Complex, Manipal-576104, Karnataka, India *Presenting author: [email protected]
Plants are immobile organisms that use various light signals to acclimatize their growth and development to the inconsistent environmental conditions. He-Ne is a powerful source of light that has widely been applied in the field of agriculture and it accelerates seed germination rate, crop yield, biomass and resistant to stress conditions. The laser excites the biological molecules, leading to photochemical transformations at molecular level. Therefore, the aim of this study is to understand the influence of He-Ne laser on endogenous phytohormone level and yield characters of brinjal on in vitro and in vivo conditions. In the present study, the seeds of brinjal (Solanum melongena L.) var. Mattu Gulla were irradiated with single exposure of He–Ne laser at different doses of 20, 25 & 30 J/cm2 and germinated aseptically on MS basal medium. Various physiological and growth parameters of the seedlings established in in vitro & in vivo conditions were estimated and significant enhancement was observed with respect to morphological and yield characteristics. The basis of stimulation process by red light could be the synergism between laser beam and the phytochrome receptors. Phytochrome controls the seed germination via regulation of two plant hormones, abscisic acid (ABA) and gibberellic acid (GA3). The hormones were extracted from the control and laser irradiated seedlings, quantified using HPLC and the endogenous level of the hormones revealed the antagonistic regulation between ABA and GA3. The results of the present study will be presented and discussed.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 195 Session B : Signaling and Phototransduction
PB162 (IPPC0885) Relevance of WUE in improving growth rates in rice (Oryza sativa L.) lines with comparable root and leaf area A.B. Priyanka*, D. Pushpa, M. Pratibha, S. Rajesh Kumar, B. Pooja, B.R. Raju, M. Udayakumar and M.S. Sheshsshayee Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus, Bangalore-560065, Karnataka, India *Presenting author: [email protected]
Rice is the highest user of irrigation water. With the increasing demand for fresh water and erratic monsoon due to climate change, rice production is fast becoming unsustainable. Thus rice cultivars with reduced water requirement need to be developed through pyramiding drought adaptive traits. Although water use efficiency is considered as an important trait, its relevance in reducing water requirement and improving yield has remained equivocal. Here we report that WUE is indeed an important determinant of growth rates when other integral traits like root and leaf area are optimised. This hypothesis was verified using 30 rice genotypes with similar leaf area and root weight but differing in Δ13C, a surrogate for WUE. Trait diversity among these genotypes was revalidated and four contrasting genotypes with significant variation in Δ13C were selected. These lines were raised in pots maintained at 100% FC and 60% FC by gravimetry. A strong positive correlation between WUE and TDM indicated the relevance of WUE. Based on the influence of physiological parameters on variability in WUE, genotypes can be classified as conductance and capacity type. The capacity types possess superior efficiency of photosynthesis and hence can produce more biomass at a given water use. The genotype R-44 emerged as such a capacity type. This genotype showed superior carboxylation efficiency based on CO2 response curves and a better mesophyll conductance as evidenced by chlorophyll fluorescence kinetics. The study provided convincing experimental evidences to prove the relevance of WUE.
PB163 (IPPC0889) Proteomic analysis of calcium signallome under dehydration stress in rice Deepti Gupta1, Yogita Rai1* and Niranjan Chakraborty2 1Department of Biotechnology, TERI University, Vasant Kunj, New Delhi-110070, India 2National Institute of Plant Genome Research, Aruna Asif Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Calcium is a ubiquitous secondary messenger involved in various stress signalling pathways in plants. It has been reported that exogenously applied calcium leads to alleviation of dehydration stress in plants. The present work was done to emphasise the role of exogenous calcium by using a calcium channel blocker (Lanthanum chloride) on physiology of dehydration stressed plants. 2-D coupled with MS/MS was used to generate a calcium signallome map of dehydration stressed plant. The proteomic investigation led to the identification and characterization of novel protein/s which have been grouped into transcription factors, metabolism, signalling and structural proteins. A novel protein having its role in plant development but no reports under abiotic stress response has been identified and characterized. This study presents significant step towards understanding of calcium signalling network under water deficit stress in rice.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 196 Session B : Signaling and Phototransduction
PB164 (IPPC0990) HOX 10 – A homeodomin transcription factor controlling root length in rice Rajesh Kumar Singal1*, K. Sumanth Kumara1, H.R. Sowmya1, M.P. Rajanna2, R.P. Sharma3, T. Mohapatra4, N. Sarala5, S. Robin6, A.K. Singh7 and M.S. Sheshshayee1 1Department of Crop Physiology, University of Agricultural Sciences, GKVK, UAS Bengaluru-560065, Karnataka, India 2Zonal Agricultural Research Station, College of Agriculture, Mandya 3National Research Centre on Plant Biotechnology, IARI, New Delhi-110012, India 4Central Rice Research Institute, Cuttack-753006, India 5Directorate of Rice Research, Rajendranagar-500030, Hyderabad, India 6Department of Rice, Tamil Nadu Agricultural Research Institute, Coimbatore, India 7National Bureau of Plant Genetic Resources, New Delhi-110012, India *Presenting author: [email protected]
Mutants with single nucleotide variations created by treating seeds with EMS form an excellent resource for functional genomics in plants. An upland rice cultivar, Nagina 22 was mutagenized and a set of mutagenized lines were screened for variations in root traits and WUE using Δ13C and other standard protocols. One high root volume and one low root volume mutant were identified and advanced to stabilize the genotype. These mutants along with the N-22 were used for targeted re-sequencing of around 120 root and photosynthesis related genes by NGS technology. A non-synonymous nucleotide variation was noticed resulting in a change of guanine to thymine in a homeodomin transcription factor HOX 10. These mutants were extensively characterized for physiological alterations under well watered (WW, 100% FC) and water limited (WL, 70% FC) conditions done by using gravimetric procedure. The high root mutant maintained high root volume, low root length and significantly higher water relations as well as canopy gas exchange characteristics under WL conditions compared to the wild type and the shallow rooted mutant. To further investigate the genetic basis of the HOX 10 mutant allele, a MUTMAP population was generated by crossing the high root mutant (N22_BADT_392_9_1) with the wild type. Selective genotyping of F2 MUTMAP progeny differing in root traits was performed by sequencing the HOX 10 gene. The HOX 10 mutant allele was found in all the 10 low root length F2 progeny that had high root volume. The results clearly demonstrated the role of HOX 10 transcription factor in regulating root architecture by influencing root length and proliferation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 197 Session B : Signaling and Phototransduction
PB165 (IPPC0991) Detoxification of reactive cytotoxic compounds by Aldo-keto reductases (AKR’s) improves seed viability and seedling vigour in rice K.N. Nisarga*, S.V. Ramu, N. Ashwini, M. Udaykumar and M.B. Raju Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus, Bangalore-560065, Karnataka, India *Presenting author: [email protected]
Seeds deteriorate during storage and in this process, reactive oxygen species (ROS) mediated lipid peroxidation leads to generation of reactive carbonyl compounds (RCC’S) and non-enzymatic modifications of proteins through maillard and amodari reactions. This leads to early loss of seed viability and vigour. Therefore, managing these compounds has relevance in improving the seed viability and seedling vigour. Towards this, an attempt was made to assess the variation in lipid peroxidation induced loss of seed viability through accelerated ageing technique in selected rice genotypes. The results showed variation in level of cytotoxic compounds such as melondialdehyde (MDA), Methyl glyaoxal (MG), Miallard and amodari reaction products with longer seed viability genotypes showing less compared to short viable genotypes. NADPH dependent dehydrogensae activity was found to be significantly low in short viable genotypes. Further, a significant inverse relationship was found between the level of cytotoxic compounds and seed viability and seed germination. This clearly suggests that, the level cytotoxic compounds in seeds determine the seed longevity. Aldo-keto reductases, are known to detoxify the cytotoxic compounds and accordingly, AKR1 transgenic was developed in a short viability rice geneotype (Telahamsa) to check the relevance of AKR in regulating seed viability. AKR1 transgenics when subjected for ageing treatment, showed improved seed germination, longer seed viability, vigour with reduced level of cytotoxic compounds compared to the wild type. This study therefore, provides evidence that, detoxification of reactive cytotoxic compounds is important in improving the seed viability and AKR’s proved to be effective in regulating the cytotoxic compounds.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 198 Session C : Genomics, Epigenomics and Regulation Biology
PC166 (IPPC0093) Proteomic, metabolomic and ionomic analysis of a halophyte, Suaeda sp. to understand its molecular adaptation Wungrampha Silas1*, Sneh Singla-Pareek2, Jayant Tripathy3 and Ashwani Pareek1 1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India 3School of Environmental Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Halophytes, an extremophile plant group, represent about 2% of the plant kingdom. They can grow at salt concentrations higher than 200 mM. The ability of halophytes to grow in saline soils has enabled them to flourish well in diverse geographical areas. Research focused on elucidating mechanism of salinity tolerance in halophytes is recently picking up its pace. Molecular analysis to understand the mechanism of adaptation and further improve their yields is underway. Proteomics, metabolomics and ionomics of a succulent halophyte, Suaeda sp. grown at the salt extraction site of Sambhar Lake, Rajasthan were analysed to understand the molecular adaptation for it to grow in that harsh environment. The plant sample was harvested in the month of June, the driest season of the year, in the lake area when the salinity is at its highest. And August, which is just after the monsoon and the site is covered with all types of vegetation. The samples were harvested at various intervals during twenty four hour period to check the circadian molecular expressional changes occurring in the plant.
PC167 (IPPC0119) Exploring the roles of cysteine-rich secretory proteins in bacteroid differentiation and nitrogen fixation in peanut nodules Senjuti Sinharoy1*, Ivone Torres-Jerez2, Kurtis Cressman3, Josh Clevenger4, Peggy Ozark Atkins4, Brian Scheffler5 and Michael Udvardi2 1Department of Biotechnology, University of Calcutta, Kolkata-700037, India 2The Samuel Roberts Noble Foundation, Plant Biology Division, Oklahoma, USA 3Oklahoma State University, OK 74074, USA 4University of Georgia, College of Agriculture and Environment Science, UGA 5USDA-ARS JWDSRC, Stoneville, MS *Presenting author: [email protected]
Legumes develop specialized organs called nodules for NF symbiosis with rhizobia, which are released into nodule cells via endocytosis, forming a unique organelle called the symbiosome. Inside the symbiosome, rhizobia differentiate into NF bacteroids. Infected nodule cells each accommodate thousands of bacteroids. Plant cells control bacteroid differentiation, which in some legume species involves nodule-specific cysteine-rich (NCR) peptides These peptides promote elongated/Y-shaped symbiosome formation. Peanut (Arachis hypogaea) has a very high nitrogen fixation capacity by the virtue of forming spherical terminally-differentiated bacteroids. We have performed RNA sequencing analysis of peanut nodule developmental to understand the molecular mechanism behind peanut spherical symbiosome formation. The peanut genome encodes around 130 cysteine-rich secretory proteins (CRSPs) that are longer (around 300-350 amino acids) than NCRs and possess a signal peptide and a pathogen-related (PR1) cysteine-rich domain, unlike the NCRs. We are exploring the possibility that CRSPs play dual roles in the control of symbionts and pathogens in peanut.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 199 Session C : Genomics, Epigenomics and Regulation Biology
PC168 (IPPC0141) Genome-wide survey and expression analysis of genes encoding seed storage proteins in chickpea (Cicer arietinum L.) Subodh Verma*, Chandra Kant and Sabhyata Bhatia National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Chickpea seeds are one of the major sources of plant protein for poor and vegetarian population especially in developing countries. However, considering the importance of chickpea for human nutrition and agricultural commodities, the molecular mechanisms that program chickpea seed development are not well understood. In mature chickpea seeds, storage proteins are the major storage compounds and accumulate during the late phases in the process of seed development. To understand the regulatory machinery of seed storage proteins (SSPs) synthesis and accumulation, a genome wide analysis of genes encoding SSPs was carried out from three legumes including chikcpea, medicago and soybean. Publicly available transcriptome data of different chickpea tissues was further utilized for in silico expression profiling of SSP encoding genes and genes encoding transcription factors (TFs). Moreover, co-expression analysis of SSPs and TFs resulted in identification of transcription factors having expression profiles similar to SSPs. These TFs included members of B3, bHLH, bZIP and C3H families. This is the first study of genome-wide identification of SSP encoding genes and their expression pattern during seed development in chickpea. This comprehensive study will provide a useful resource to explore the regulatory networks involved in SSP synthesis and accumulation in chickpea developing seeds.
PC169 (IPPC0245) Genome-wide comparative profiling of heat shock proteins and heat shock factor during multiple abiotic stresses reveals superimposed and stress specific adaptational responses in Pennisetum glaucum and Oryza sativa Saurabh Pandey* and M.K. Reddy Plant Molecular Biology Lab, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Plants possess a flexible and elaborate nexus of heat shock protein (HSPs) and heat shock factors (HSFs) that are widely involved in cellular response to multiple abiotic stresses besides heat. HSPs are accountable for protein folding, assembly, translocation, protein intracellular localization, secretion and degradation in a broad array of cellular processes in normal and stressed cells. HSPs are transcriptionally activated by HSFs that are transcription factors involved in response to different abiotic stresses. To illustrate the functional regulation of Pennisetum glaucum HSP and HSF genes, we analyzed the changes in their global expression profiles in response to heat, salt, cold and drought stresses in seedlings. Comparative analyses revealed that while high resemblance existed in expression of most HSPs and HSF with an overlapped response and regulation patterns during different stresses, some genes showed notably stress- specific response. Interpretation of microarray data analysis identified several functionally correlated genes comprising of stress up-regulated known and new genes that may be implicated in heat and other stresses. Based on the findings on collateral analysis of stress associated variation in transcripts of selected HSPs, genome wide microarray profiling and co-expression analysis, present study is a step ahead in comprehending the mechanism underlying stress response in Pennisetum glaucum. Moreover, heat-responsive gene profiling differed largely from those under drought, cold and salt stresses and drought treatment was more effective in up-regulating HSF expression. Results advocate that the conserved expression of HSPs and HSFs in response to multiple stresses may highlight their role as important components facilitating cross-talk between various stress-related signal transduction pathways.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 200 Session C : Genomics, Epigenomics and Regulation Biology
PC170 (IPPC0260) Specificity of interactions between ARFs and ARF-GEFs during vesicle formation at different membranes in the cell Manoj Kumar Singh1*, Sandra Richter1, Hauke Beckmann1, Marika Kientz1, Nadine Anders2, York Stierhof1, Ulrike Mayer1 and Gerd Juergens1 1Center for Plant Molecular Biology, Developmental Genetics, University of Tübingen, Germany 2Department of Biochemistry, University of Cambridge, Cambridge CB2 1QW, United Kingdom *Presenting author: [email protected]
Transport of various biomolecules between different subcellular compartments through vesicle trafficking is essential for optimal growth of plants and for their ability to adapt to different biotic and abiotic stresses factors. Formation of a vesicle from a donor compartment is regulated by the ARF family of monomeric GTPases. At the site of vesicle budding, GDP- bound ARF GTPases’ are recruited from the cytosol to the membrane and become activated by replacing the bound GDP with a GTP. This activation process requires interaction of ARFs with ARF guanine nucleotide exchange factors (ARF-GEFs). Once GTP-bound, ARFs recruit the coat machinery, which is necessary for formation and scission of the cargo- containing vesicles. The hydrolysis of bound GTP to GDP inactivates the ARF and results in dissociation of coat proteins from the vesicle, allowing its fusion with the acceptor compartment. Arabidopsis thaliana genome encodes nine ARF GTPases and eight ARF-GEFs. Functional studies have revealed the specific roles of different ARF-GEFs in secretion, recycling of the auxin efflux carrier protein PIN1, trafficking to the cell plate during cytokinesis, and vacuolar transport. Although ARF-GEFs have been studied extensively, relatively little information is available about the role of different ARF GTPases and, more importantly, the specificity of ARF-GEF interactions with ARFs at different subcellular compartments. Our findings from biochemical, genetic and microscopic analysis of transgenic Arabidopsis plants will be presented and discussed.
PC171 (IPPC0284) Study on role of Arabidopsis Methyl CpG binding domain protein 4 (AtMBD4) in phosphate starvation Adwaita Parida, Amrapali Sharma* and Arun Sharma Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India *Presenting author: [email protected]
Methyl CpG Binding Domain proteins are a special class of protein that recognize and bind to methylated DNA. In Arabidopsis, mutation in the Methyl CpG Binding Domain protein 4 gene (AtMBD4) shows more lateral root development as compared to wild type plants. Transcriptome analysis of atmbd4 mutant using microarray reveals that this phenotype is due to up-regulation of many genes involved in phosphate starvation response. Plants grown in phosphate deficient condition also show a similar phenotype as that of atmbd4 mutant. Yeast two hybrid screening using AtMBD4 as bait; found a Ubiquitin conjugating enzyme 36(AtUBC36) that interacts with AtMBD4. AtMBD4:CFP and AtUBC36:YFP fusion proteins localized in the nucleus. Further in vivo interaction between these two proteins was analyzed in plant cell using Florescence Resonance Energy transfer (FRET). Both the proteins were co-localized in the nucleus and showed a very good FRET efficiency that confirmed their association. The mutant atubc36 also showed the similar root phenotype with more lateral roots like atmbd4. Both the genes were up-regulated during phosphate starvation. Moreover, both atmbd4 and atubc36 mutants were found to accumulate more inorganic phosphate and anthocyanin as compared to wild-type plants. Our results clearly suggest that these two proteins interact with each other and perform an important role during phosphate starvation response.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 201 Session C : Genomics, Epigenomics and Regulation Biology
PC172 (IPPC0301) The evolution of pitchers in Nepenthes khasiana: how many and which genes does it take to make a pitcher? Jeremy Dkhar*, Yogendra Bhaskar, Andrew Lynn and Ashwani Pareek School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India *Presenting author: [email protected] A structural phenomenon seen in certain lineages of angiosperms that has captivated many scholars including Charles Darwin is the evolution of plant carnivory. We now understand why carnivory evolved but how carnivorous plants acquired these attributes remain a mystery. In an attempt to understand the evolution of pitchers, a carnivorous trait found in three flowering plant families, we de novo assembled for the first time the transcriptome sequence of Nepenthes khasiana, an insectivorous plant endemic to the state of Meghalaya, North-east India. We generated more than 180 million reads using Illumina sequencing from the highly specialized N. khasiana leaf comprising the leaf base, tendril and the different parts/zones of the pitcher tube viz. digestive zone, waxy zone and lid. We then mapped respective reads into the de novo assembled reference transcriptome to quantify transcript abundance along the N. khasiana leaf. We detected highest number of expressed transcript in the tendril followed by waxy zone, digestive zone, lid and leaf base. Hierarchical clustering of differentially expressed genes indicated functional relationship and similar cellular processes underlying the leaf base and pitcher tube, thereby implying that the Nepenthes pitcher is indeed a modified leaf. Taking cues from SEM and LM photomicrographs of the five distinct parts/zones of the N. khasiana leaf, we examine the expression pattern of genes involved in stomatal and trichome development as well as leaf polarity specification genes. Our findings imply that alterations in the expression of key regulatory genes involved in leaf development probably led to the evolution of Nepenthes pitchers.
PC173 (IPPC0310) Integrating RNA transcriptome wide and microRNA analyses for the identification of molecular regulators associated with high salt tolerance in Sesuvium portulacastrum (L.) Ganesh Nikalje1*, Ashish Kumar Srivastava2, Gaurav Sablok3, Tukaram Nikam1 and Suprasanna Penna2 1Department of Botany, Savitribai Phule Pune University, Pune-413007, India 2Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai, India 3Plant Functional Biology and Climate Change Cluster (C3), University of Technology Sydney, Sydney *Presenting author: [email protected] Sesuvium portulacastrum, a member of Aizoaceae family, is endowed with its ability to complete its life cycle under high salinity which is associated with regulation of cellular energetics and redox homeostasis. The present study was conducted to identify transcriptional mediators related to salt adaptation. On the basis of post-germination phenotyping, 100 and 250 mM NaCl concentrations were selected at which, growth was either comparable to control or reduced by 50%, respectively. The growth reduction at 250 mM was concomitant with higher Na+ accumulation at 10 d; although, at 24 h after stress, Na+ accumulation was equally high at both the salt concentrations. The activation of enzymatic cascade was seen only at 250 mM. At 24 h treatment, total RNA was isolated from root and RNAseq was performed at Illumina HiSeq platform. A total of 189 million reads were assembled into high quality 104843 unigenes and their functional annotation revealed key salt responsive genes. To link this with micro regulators of RNA mediation regulation, a set of conserved miRNAs (miR156, miR159, miR162, miR164, miR167, miR169, miR172, miR390, miR395, miR399 and miR838) and their expression was measured using stem-loop RT-PCR. A total of 33 targets were identified using assembled transcriptome and their expression data confirmed that transcriptional changes were more pronounced at 250 mM NaCl. This suggested that observed effects are more closely related with osmotic components. Thus, present study highlights the concentration dependent effects on growth and identifies key genetic elements associated with salt adaptation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 202 Session C : Genomics, Epigenomics and Regulation Biology
PC174 (IPPC0316) Phylogenomic analysis of AP2/ERF family genes in sorghum Sushree Sangita1, Supriya Mathur1*, Aleena Francis2, Rita Sharma2 and Manoj Sharma1 1School of Biotechnology, 2School of Computational & Integrative Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Sorghum bicolor is the fifth most important cereal crop in the world. It not only provides food, feed and fiber but is also emerging as a leading biofuel crop. It is a low input crop with high water-use efficiency and potential to recover growth after drought stress. However, changing environmental conditions pose a serious threat to plant growth and productivity. Therefore, it is imperative to understand the mechanism of stress adaptation and identify potential candidates for breeding improved varieties in Sorghum. The members of AP2/ERF family transcription factors have particularly emerged as important candidates for engineering stress tolerance in crop plants. They are characterized by AP2/ERF DNA binding domain and have been further classified into three subfamilies including AP2, ERF and RAV. To elucidate their role in plant development and stress response in sorghum, we performed genome-wide identification and phylogenomic analysis of AP2/ERF family genes in Sorghum. Using HMM and NCBI-CDD analysis, we have identified more than 175 ERF family genes in sorghum. We present here the results of phylogenetic analysis, structural characterization and expression analysis of candidate AP2/ ERF genes. Expression analysis of the genes was performed at using publicly available transcriptomic data developmental stages and in response to abiotic stress treatments.
PC175 (IPPC0336) microRNAs in chickpea (Cicer arietinum L.) root nodulation Chandra Kant* and Sabhyata Bhatia National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India *Presenting author: [email protected]
Chickpea (Cicer arietinum L.) is widely grown across the Indian subcontinent. In symbiotic association with Mesorhizobium ciceri root nodules are formed that play important role in the farming system by fixing atmospheric nitrogen. miRNAs have emerged as an important regulatory component in a number of developmental processes including legume- rhizobia symbiosis. In order to understand the role of miRNAs in root nodulation a smallRNA library from root nodules of chickpea was sequenced using the Illumina GA II X platform. Conserved miRNAs were identified based on their similarity to miRNA of other plant species present in miRBase, and putative novel miRNAs were identified based on potential hairpin loop forming precursors from available plant databases. Presence of miRNAs were validated by using northern blotting and stem loop RT-PCR. In-silico expression profile of miRNAs in nodule in comparison to other tissues and target identification revealed a set of miRNAs that may have imperative role in nodule development and function, which were further validated by Quantitative Real Time PCR. The conserved and putative novel miRNA identified in this study will help reveal intricacies of chickpea-Mesorhizobium symbiosis.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 203 Session C : Genomics, Epigenomics and Regulation Biology
PC176 (IPPC0357) Expression and protein: protein interaction analysis of FCS-LIKE ZINC FINGER 17 and 18 of Arabidopsis thaliana Muhammed Jamsheer K, Manvi Sharma*, Chanchal Thomas Mannully and Ashverya Laxmi National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
The FCS-LIKE ZINC FINGER (FLZ) family is a plant-specific family of zinc finger proteins characterized by the C2-C2 FCS-LIKE ZINC FINGER (FLZ) domain. The FLZ domain acts as a protein-protein interaction module. The FLZ domain containing proteins are identified from all sequenced spermatophytes out of which the Arabidopsis thaliana genome possesses 18 FLZ domain genes. Out of theses 18 genes, FLZ17 and FLZ18 are tandem duplicates without any sequence divergence. Both these proteins are highly diverged from other members of this gene family. FLZ17/18 is localized in both nuclei as well as cytoplasm and interacts with many other members of the same class of zinc finger. Moreover, the Y2H analysis identified that they interact with many regulators of processes like flower development, photosynthesis, photomorphogenesis etc suggesting the possible function of these genes in these processes. Consistent with this observation, FLZ17/18 found to be profusely expressed in flower and silique stages and during the vegetative- to-reproductive phase transition. Similarly, the expression of these genes was found to be differentially regulated by various biotic as well as abiotic stresses. The genes were found to be differentially regulated by hormones involved in biotic and abiotic stresses. Taken together, the present study gives a comprehensive account of the transcriptional regulation and protein: protein interaction properties of FLZ17/18 which will be helpful in the molecular and functional characterization of these genes.
PC177 (IPPC0412) Variability for root traits in sunflower hybrids and inbreds Lakshmi Prayaga*, P. Lakshmamma and C. Sarada Department of Plant Physiology, Indian Institute of Oilseeds Research, Rajendranagar, Hyderabad-500030, India *Presenting author: [email protected]
Fifty four inbreds and 8 hybrids of sunflower were studied for variability in root traits by growing them in polybags (90 cm length and 40 cm diameter) of 120kg soil capacity @ 2 plants per bag for 65 days during rabi. FYM was mixed with soil in the ratio of 1: 5 (v/v). Plants were irrigated as and when required. Observations were recorded at the time of harvest. Wide genetic variability was observed for all the parameters studied. Hybrids recorded higher root weight, volume, length, total dry matter (TDM), leaf area, leaf number compared to inbreds. Root weight showed strong positive relationship with TDM (0.77) and leaf area (0.62) compared to root volume (0.74 & 0.61) and root length (0.62 & 0.55). Among the root traits, root weight and volume showed good correlation (+0.89) compared to their relation with root length. Root traits did not show any relation with SCMR and SLA (specific leaf area). A selection index was computed using principal component analysis by taking into consideration root characters along with TDM and leaf area. Based on the index value, hybrids DRSH-1, NSFH-1001 and inbreds 106A, 335 A, AKSFI-42-I, DS-2, 298-R, CSFI-5075, 856-R, R-7, CMS-335A were found promising.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 204 Session C : Genomics, Epigenomics and Regulation Biology
PC178 (IPPC0417) Occurance and arrangement of three mutisubunit inorganic carbon transporters in cyanobacteria Vandana Tomar*, Gurpreet Kaur Sidhu, Panchsheela Nogia, Rajesh Mehrotra and Sandhya Mehrotra Biological Sciences, Birla Institute of Technology and Science, Qtr.28/1 Pashchim Marg BITS, Pilani-333031, India *Presenting author: [email protected]
The oxygenase reaction catalyzed by Rubisco became an issue only after the evolution of the oxygenic photosynthesis in cyanobacteria. Several strategies were developed by autotrophic organisms to increase oxygen levels in the viscinity of Rubisco to maximize its net carboxylation rate. One of the crucial advancements in this context was the development of more efficient inorganic carbon transporters which could help in increasing the influx of inorganic carbon (Ci) at the site of CO2 fixation. A survey was done to find out the genes encoding for cyanobacterial Ci transporters in twenty cyanobacterial phyla with respect to transporters present in Gloeobacter violaceous PCC 7421, an early diverging cyanobacterium. An attempt was also made to correlate the prevalence of the kind of transporter present in the species with its habitat. Basically two types of cyanobacterial inorganic carbon transporters exist i.e., bicarbonate transporters and CO2 uptake systems. The transporters also show variation in context to their structure as some exist as single gene encoded proteins (BicA and SbtA), while others exist as multisubunit proteins (BCT1, NdhI3 and NdhI4). The phylogeny and distribution of the former have been extensively studied and the present analysis provides an insight into the latter ones. The in silico analysis of the genes under study revealed that their distribution was greatly influenced by the habitat and major environmental changes in the course of their evolution viz., great oxidation event. Knowledge of the evolution of these transporters could be a step towards selection of suitable candidates for plant transformation.
PC179 (IPPC0458) Transcriptome modulation in Arabidopsis natural variations under arsenic stress Tapsi Shukla1*, Smita Kumar2, Deepika Sharma1, Deepika Lakhwani1, Ria Khare1, Mehar Asif1 and Prabodh Trivedi1 1Molecular Biology, CSIR- National Botanical Research Institute, Rana Pratap Marg, Lucknow-226001, India 2University of Lucknow, Lucknow, India *Presenting author: [email protected]
Heavy metal contamination primarily due to the industrial processes, agricultural practices, mining and other anthropogenic activities has resulted in severe threat to environment and human health. Among other heavy metals, arsenic (As) is most toxic to biota even at low concentration and is considered to be a potent carcinogen. In view of As toxicity there is an increasing concern about the molecular mechanisms underlying the uptake, transport and accumulation of As in plants. In recent years, considerable attention has focused on the use of Arabidopsis natural variations to understand adaptation and response to environmental changes. Here, we analysed the response of Arabidopsis accessions towards As stress. Differential response in terms of phenotypic variation (root length) was observed in the accessions exposed to different concentrations of As. On the basis of reduction in the primary root length, nine accessions were categorized as tolerant, moderate and sensitive for As(III) stress. The accessions Col-0, Koch-1, Sij-2 were tolerant (< 55% reduction in primary root length) whereas Borsk-2, Altneb-2, Sij-1 were moderate (> 55-60% reduction) and Ciste-2, Slavi-1, Ws were sensitive (> 60% reduction) towards As(III) stress. To study the contrasting response of different accessions, genome- wide expression analysis using most tolerant and sensitive accessions was carried out. A group of defence and stress- responsive genes, transporters, and regulatory genes showed differential expression in the contrasting accessions. The modulation in the expression pattern of the genes that showed the highest contrast in the microarray was further validated by qRT-PCR. Thus, the study provides an insight about the genes and the molecular mechanisms involved in arsenic stress response/ adaptation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 205 Session C : Genomics, Epigenomics and Regulation Biology
PC180 (IPPC0470) HY5-dependent differential expression of genes involved in flavonol biosynthesis under low temperature and light Chitra Bhatia1*, Ashutosh Pandey2 and Prabodh Trivedi 1Plant Genomics Lab, CSIR-National Botanical Research Institute, Lucknow-226001, India 2National Agri-Food Biotechnology Institute, Punjab, India *Presenting author: [email protected]
Light and temperature are two most important factors affecting plant growth, development by various biological processes. Flavonoids are a class of secondary metabolites produced by the plants in response to stress conditions through phenylpropanoid pathway. Studies suggest that biosynthesis of flavonoids is regulated by light through various regulatory factors as well as proteosomal machinery. LONG HYPOCOTYL 5 (HY5), a bZIP transcription factor, binds directly to the promoters of light-inducible genes and plays important regulatory role in flavonoid biosynthesis. During dark conditions, HY5 is degraded in the plant via proteasome-mediated degradation involving CONSTITUTIVE PHOTOMORPHOGENESIS 1 (COP1), RING E3 ubiquitin ligase. Previous reports suggest that during low temperature and light conditions, enhanced biosynthesis of anthocyanins is regulated through HY5-dependant manner. However, very little information is available on effect of low temperature and light on regulation of flavonol biosynthesis which shares the same pathway as anthocyanins. In this study, we analyzed expression of regulatory (MYB11, MYB111 and MYB12) and structural genes (CHS, CHI, F3H and FLS) involved in flavonol biosynthesis under different light qualities in wild type, hy5-215 and cop1-4 mutants of Arabidopsis thaliana. Similarly, expression analysis of these genes in low temperature conditions under white light was also performed. Our results suggest induction of regulatory and structural genes involved in flavonol biosynthetic pathway in response to light as well as low temperature conditions in HY5- dependant manner.
PC181 (IPPC0475) Comparative analysis of methylome under abiotic stress in foxtail millet cultivars [Setaria italica (L.) P. Beauv.] Garima Pandey*, Chandrabhan Yadav and Manoj Prasad Plant Molecular Biology, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Foxtail millet is a C4 Panicoid crop with a relatively small genome, short life-cycle and is closely related to several biofuel crops. Apart from this, it is a potential abiotic stress tolerant crop which categorizes it as an excellent model system to study the stress induced gene regulation mechanisms. DNA methylation is one of the important mechanisms related to gene regulation at epigenetic level. As methylation based stress regulatory mechanism is poorly understood in foxtail millet, which is naturally adapted to abiotic stress, we aimed to study DNA methylation level in two contrasting cultivars of foxtail millet (i.e., tolerant cv. IC403579 and sensitive cv. IC480117) under salt stress condition using Methylation Sensitive Amplification Polymorphism (MSAP) technique. Reduced level of DNA methylation was observed in tolerant cultivar as compared to the sensitive cultivar. Total 69 genes showed differential methylation behaviour in both the cultivars. Majority of which corresponds to transposase related genes which are responsible for activation and transposition of transposons in the genome. Some of the transposase related genes were subjected to bisulfite sequencing for further validation. Along with this expression analysis of demethylases showed significant upregulation in tolerant cultivar under salinity stress. These results indicate that on exposure to salinity stress, demethylation of genome by demethylases might lead to activation of stress related genes which may be one of the probable reasons for stress tolerance behaviour in foxtail millet cultivar. Further, functional characterization of stress-induced differentially methylated genes will provide profound understanding of abiotic stress tolerance mechanism in millets.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 206 Session C : Genomics, Epigenomics and Regulation Biology
PC182 (IPPC0507) MicroRNAs: Small nodes of stress management in root apical meristem in chickpea Hitaishi Khandal* and Debasis Chattopadhyay Plant Molecular Biology, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Root growth is regulated by cell division at the root apical meristem. Structure of root apical meristem of Fabaceae plants is notably different from that of Arabidopsis. Histological studies have shown that unlike a closed structure of RAM in Arabidopsis, RAM of legume plants exhibits a basic-open structure. MicroRNAs have emerged as regulators of many key biological functions in plants including stress management. To identify key miRNAs involved in abiotic stress management in root meristem of chickpea, we used deep sequencing to compare miRNA populations among five small RNA libraries prepared from control and abiotic stress treated chickpea root meristem (~2 mm). A total of 8,03,259 high quality reads were generated. 458 unique sequences were mapped 100% to the chickpea genome. Statistical analysis on transcript abundance of predicted annotated and putative miRNAs revealed that almost 90% of them were differentially regulated by PEG induced dehydration and salinity treatments at early and late stages. We identified a total of 284 putative miRNA sequences out of which 224 were already annotated in miRBASE dataset which belongs to 195 unique families, while rest of 60 sequences were categorized as predicted putative miRNAs. The chickpea miRNAs targeted a wide range of mRNAs involved in diverse cellular processes, including transcriptional regulation (transcription factors), protein modification, signal transduction, and metabolism. This study reveals a comprehensive set of conserved and novel miRNAs along with their expression patterns and putative targets in chickpea, and provides a framework for understanding regulation of developmental processes and stress tolerance mechanisms in legumes.
PC183 (IPPC0517) Proteomic analysis of chickpea (Cicer arietinum L.) reveals dynamic plasma membrane architecture Praga Barua*, Nilesh Lande, Subhra Chakraborty and Niranjan Chakraborty National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Plant cells contain numerous membrane systems that are specialized to perform particular functions. The plasma membrane (PM), in particular, is an organized system that serves as physical barrier and as a communication interface with the extracellular environment. Most importantly, the processing of cell signaling reactions that are involved in responses to biotic and abiotic stressors occurs in the plasma membrane. The multivariate role of plasma membrane demands its proteome to be highly dynamic, comprising a large number of proteins with highly varied function. Therefore, characterization of the plant plasma membrane proteome can provide new insights into its multivariate functions. In an attempt to understand the complexity of plant plasma membrane proteins (PMPs), we developed a proteome reference map of a food legume, chickpea. Plasma membranes were isolated from 3-week-old seedlings and purity of the fraction was confirmed by immunoblot screening. The proteome was constructed using plasma membrane enriched fraction using 1-DE technique. Analysis of 1-D bands as well as in-solution tryptic digests of PMP led to the identification of more than 2000 proteins including well known intrinsic PMPs such as aquaporins, H+ATPase etc, as well as many predicted and uncharacterized proteins. This comprehensive overview of the proteome would provide a basis for future comparative proteomics efforts for other crop species. Our future efforts would focus on identifying differentially regulated proteins under stress conditions, which may help in the long-term efforts to develop transgenic crops with improved stress tolerance.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 207 Session C : Genomics, Epigenomics and Regulation Biology
PC184 (IPPC0549) Understanding the epigenetic regulation of molecular memory of infection in Arabidopsis Zeeshan Zahoor Banday*, Sadan Sharma and Ashis K. Nandi School of Life Sciences, Jawaharlal Nehru University, New Delhi-110 067, India *Presenting author: [email protected]
Immobility poses a challenge to plants. Plants live in complex milieu where they interact with a wide range of organisms. These interactions can be beneficial or detrimental. Plants respond to pathogen attack by activating local as well as systemic defenses that limit the growth and spread of pathogen. The reduced in systemic immunity 1 (rsi1) identified by the genetic screen of EMS mutagenized wild-type Arabidopsis, fails to activate systemic acquired resistance (SAR). RSI1 is allelic to Flowering locus D (FLD). The mechanism of FLD mediated SAR activation, however, is not known. FLD is known to function as a part of transcriptional co-repressor complex. The protein interacting partners of FLD in the co- repressor complex remain largely unidentified. To elucidate the possible mechanism of FLD mediated SAR activation, we screened Arabidopsis thaliana cDNA library for FLD interacting proteins (FINs) in yeast. We identified several putative interacting factors of FLD. Few of these interactors play a relevant role in defense and could be important candidates for FLD to form a functional complex with, underlining its role in epigenetic modification of SAR. Here we report the studies on one such FIN, FIN1.We show that FIN1 physically interacts with FLD. We also demonstrate the nuclear co-localization and in vivo interaction of FIN1 with FLD. The FIN1 T-DNA insertion mutants were subsequently shown to be compromised in SAR. Understanding the mechanism of SAR regulation will have far-reaching benefits both for the improvement of crop yields and human health.
PC185 (IPPC0555) Identification of RSI1 target for systemic acquired resistance in Arabidopsis Ashis Nandi, Deepjyoti Singh* and Vijayata Singh School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Plants are under attack of large number of pathogens. However due to the presence of well elaborate immune system they remains mostly healthy. One of such immunity mechanism is Systemic Acquired Resistance (SAR); by which a partly infected by pathogen becomes highly resistance for secondary infection. Upon infection, infected tissue generates mobile signals that get transported through vasculature to rest of the plant. These signals are believed to cause epigenetic modifications to generate the state of alertness in plants. Due to this alertness the experienced plants show the SAR associated priming, i.e, higher level of defense gene expression than a naive plant. Even though a large number of mobile signals are known the molecular mechanism by which they turn into epigenetic modifier is not known. Our lab identified a mutant rsi1 (reduced systemic immunity1), through forward genetic screening of EMS treated wild type seeds of Arabidopsis (Col-0). This mutant demonstrated normal local resistance but not SAR. RSI1 is allelic to FLD. We compared the transcriptome profile of WT and rsi1 plants to identify the targets of rsi1 (TORs). By knowing RSI1/FLD is a negative regulator of transcription, we detected the genes that are specifically upregulated in SAR induced rsi1 plants. One of such genes TOR1 (Target of RSI1) shows predicted pattern of FLD/RSI1 mediated histone methylation in its promotor. The mutant phenotype of tor1.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 208 Session C : Genomics, Epigenomics and Regulation Biology
PC186 (IPPC0563) Genome-wide identification and characterization of dehydration-responsive miRNA(s) and targets in Setaria italica Amita Yadav* and Manoj Prasad Plant Molecular Biology, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected] miRNA-mediated gene regulatory networks under dehydration stress remain largely unexplored in plant including foxtail millet (Setaria italica), which is a natural abiotic stress tolerant crop. To find out the dehydration-responsive miRNAs at the global level, four small RNA libraries were constructed from control and dehydration (6h) stress-treated seedlings of two foxtail millet cultivars showing contrasting tolerance behavior towards dehydration stress. Using illumina sequencing technology, 55 known and 136 novel miRNAs were identified. Moreover, 18 known and 33 novel miRNAs were differentially expressed during dehydration stress. After stress treatment, 32 dehydration responsive miRNAs were up-regulated in tolerant cultivar IC403579 and 22 miRNAs were down-regulated in sensitive cultivar IC480117, such antagonistic behavior of foxtail millet miRNAs indicated that miRNA-mediated molecular regulation might play important roles in providing contrasting characteristics to these cultivars. Moreover, northern blot analysis and SL-qRT PCR have been performed to validate the expression patterns of some known and novel dehydration-responsive miRNAs. Differential expression behavior of few miRNA-target genes were also verified under dehydration stress treatment and miRNA- directed cleavage site was validated by RLM-RACE. One of the candidate miRNA, which is monocot-specific, showed up-regulation in sensitive cultivar and simultaneously, expression of its target was found to be down-regulated after dehydration treatment. To understand its role, particularly in dehydration stress, rice transgenic lines over-expressing its foxtail millet precursor were generated. Expression level and copy number of transgenics were analyzed and evaluation of T2 lines is in progress. This study would help in elucidating one of the dehydration-stress associated post-transcriptional regulation mechanism.
PC187 (IPPC0570) Genome wide analysis of ACGT/CRE in Arabidopsis thaliana reveals frequency of ACGT_CRE is higher than CRE_CRE Rajesh Mehrotra, Chetna Sangwan*, Sandhya Mehrotra, Vandit Gandotra and Zaiba Hasan Khan Biological Sciences, Birla Institute of Technology and Science (BITS) Pilani, Jhunjhunu-333031, India *Presenting author: [email protected]
It is inevitable to prevent the outcomes of the abiotic stress in the plants. Drought and salt stress, together with low temperature, are the major problems for agriculture because these abiotic environmental factors prevent plants from realizing their full genetic potential and affect the plant growth and productivity adversely. To combat these stresses plants have evolved mechanisms where multitude of gene expression profile changes are involved. Synergy between the cis regulatory elements and their cognate transcription factors leads to the exactitude and melange of gene expression. In plant system, 469 cis regulatory elements have been documented as per PLACE database. ACGT cis regulatory element forms the core sequence of other cis regulatory elements viz salicylic acid responsive element, abscisic acid responsive element (ABRE), core sequence of G box, light response element. Earlier study reveals that occurrence of ACGT element is one of the pivotal features of the stress responsive promoter, provided either an additional copy of the same motif or any other cis element (CREs, DREs, LREs etc.) is present. In our genome and promoter wide analysis of CREs (Cold responsive elements) focusing on the “CCGAC” and its variants in Arabidopsis thaliana, we found that the frequency of ACGT_CCGAC & CCGAC_ACGT spacers is higher than two CCGAC spacer_CCGAC in tandem. We also found that (ACGT_CCGAC) motifs with spacer lengths of 2, 9, 15, 20, 21, 26, 28 & 29 nucleotides occur at higher frequency in the promoters regions than other spacer lengths.The data obtained in this study will be presented.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 209 Session C : Genomics, Epigenomics and Regulation Biology
PC188 (IPPC0583) Trancriptional profiling during dormancy release in Apple suggests the role of epigenetic mechanism in regulating the perception of differential chilling Gulshan Kumar*, Gagandeep Singh, Mohit Swarnkar, Ram Sharma and Anil Singh Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061, H.P., India *Presenting author: [email protected]
Apple is a temperate fruit crop and low temperature plays a crucial role in floral bud dormancy and release. Therefore, comparative transcriptome profiling of dormant bud, silver tip (mid active) and green tip (active bud) of field grown apple genotype “Royal delicious” under differential chilling availability was performed using RNA-seq approach. The de novo assembly of 113,666,401 paired-end reads resulted in 47,712 contigs. The 32,408 (68%) contigs were successfully annotated by BLASTP search against non-redundant NCBI database. Higher chilling availability during dormancy period was found to be associated with more number of differentially expressed genes in transition from silver tip to green tip stage. The expression of two DAM (Dormancy Associated MADS) genes was found to be decreased with concomitant chilling acquisition, however the basic transcript abundance of these DAM genes remained higher under low chill conditions during the active growth period. Similarly, the expression of FLC-like gene was found to be decreased with chilling acquisition under high chill condition, while its expression was induced under low chill condition. In addition, the expression of predicted DRMs and CMTs involved in de novo DNA methylation, and SAM synthase (SAM, a cofactor for methyltransferases) was found to be higher under low chill conditions. Moreover, the methylation sensitive amplified polymorphism also showed higher methylation events under low chill conditions. The regulation of FLC and DAM genes has been reported to be regulated through methylation of their promoter regions. Thereby, the role of epigenetic regulation in chilling mediated dormancy release in apple is apparent.
PC189 (IPPC0624) Role of histone acetyltransferases and deacetylases in epigenetic regulation of cotton fiber development Sunil Kumar Singh, Babita*, Archana Bhardwaj, Verandra Kumar, Krishan Mohan Rai, Surendra Pratap Singh and Samir Vishwanath Sawant Plant Molecular Biology and Genetic Engineering, CSIR-National Botanical Research Institute, Lucknow- 226001, India *Presenting author: [email protected]
The cell differentiation and development largely depends upon the signals generated by dynamic control of different gene and regulatory networks acting within the cell. Histone modifications play key role in regulating the gene networks and largely contribute to the transitional alterations in expression pattern of genes. The present study aims to decipher the role of histone acetyltransferases (HATs) and deacetylases (HDACs) in epigenetic regulation of cotton fiber development. In this study, we used Anacardic acid (inhibitor of p300/PCAF class of HATs) and Trichostatin A (inhibitor of almost all HDACs, except HDA08). We performed ovule culture assays and checked the effects of inhibitors against various histone modifiers at phenotypic, biochemical and transcriptome level. We observed that Anacardic acid promotes, while Trichostatin A inhibits the fiber development in cultured cotton ovules. We identified genome wide targets of these modifiers by transcriptome sequencing. Treatment of these inhibitors induces the change in expression of various genes involved in auxin biosynthesis and response pathway. Next, we performed ChIP-seq assay to study the histone acetylation (H3K9Ac) landscape at the target genes and expressional changes due to alterations in the histone acetylation pattern by inhibitors of HATs and HDACs. Anacardic acid spraying to the developing flowers and bolls in cotton plants improves fiber yield and quality. The results suggest that histone modifiers play a crucial role in regulation of fiber cell differentiation and development.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 210 Session C : Genomics, Epigenomics and Regulation Biology
PC190 (IPPC0634) Understanding transcriptional and epigenomic regulation of abiotic stress responses in rice Rama Shankar*, Annapurna Bhattacharjee, Narayana Chevala, Rohini Garg and Mukesh Jain National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India Presenting author: [email protected]
Drought and salinity are the major environmental factors affecting the rice productivity. Various rice cultivars have been identified with evolved mechanism of abiotic stress tolerance. The comparative transcriptome analysis between stress- tolerant and -sensitive rice cultivars can give us clue about the source of genes and/or genetic variations to improve the stress tolerance of high yielding rice cultivars. We performed comparative transcriptome analysis between stress- sensitive (IR64) and -tolerant (N22 and Pokkali) rice cultivars under desiccation and salinity stresses. A large number of novel transcripts and alternatively spliced transcript isoforms were identified in the rice cultivars. Differential gene expression analysis resulted in identification of various cultivar-specific transcripts. Functional categorization of these transcripts revealed enrichment of several biological processes in N22 and Pokkali rice cultivars under stress conditions. In addition, discovery of SNPs/InDels and polymorphic SSRs were also carried out in IR64/N22, IR64/Pokkali and N22/ Pokkali. Annotation of SNPs/InDels resulted in identification of various genes harboring large-effect SNPs/InDels. These genes were found to be enriched in defense response and response to abiotic stress terms. In addition, quantitative trait loci (QTLs) harboring differentially expressed genes and large-effect SNPs/InDels have also been identified. Further, DNA methylation as a result of SNPs was discovered to find out the allele-specific methylation in the rice cultivars. Altogether, these analyses provide a comprehensive overview about the transcriptional and epigenetic regulation of stress responses, and serve as a resource to find out better genes/pathways important for abiotic stress tolerance.
PC191 (IPPC0636) Understanding the importance of KIX domain proteins Jitendra Thakur* National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Present author: [email protected]
KIX domain is found in p300/CBP and Mediator subunit Med15 and is important for protein-protein interaction. Sequence, mutational and structural studies from our and other laboratories suggest that CBP KIX has evolved from Med15 KIX domain. In fungi and metazoans it has been found that the activation domains of diverse transcription activators target KIX domain to regulate transcription of their target genes. However, not much is known about KIX domain proteins in plants. We attempted to characterize all the KIX domain proteins in Arabidopsis and rice. Interestingly, in addition to p300/CBP- and Med15-like proteins, KIX domain was also found in F-box proteins in rice and DNA helicase in Arabidopsis suggesting new roles of KIX domain in ubiquitin-mediated proteasomal degradation of protein and genome stability. In both rice and Arabidopsis, we have found more than fifty proteins interacting with the KIX domain of MED15 suggesting its involvement in diverse processes. In rice, expression analysis revealed overlapping expression of OsKIX_3, OsKIX_5 and OsKIX_7 in seeds of different stages of development, suggesting their individual or combined role during seed development. Moreover, the association analysis using data of 136 SNP loci in 23 contrasting rice genotypes and their grain length-specific phenotypic information identified three non-synonymous SNP loci in these three rice genes showing strong association with long- and short-grain differentiation. Interestingly, these SNPs are located within KIX domain encoding genomic regions. It, thus, indicates the additional importance of novel SNP loci/alleles identified in KIX domain containing genes for determining seed size in rice.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 211 Session C : Genomics, Epigenomics and Regulation Biology
PC192 (IPPC0637) Identification of novel and conserved miRNAs from extreme halophyte, Oryza coarctata, a wild relative of rice Tapan Kumar Mondal* and Showkat Ahmad Ganie National Bureau of Plant Genetic Resources, New Delhi-110012, India *Presenting author: [email protected]
Oryza coarctata, a halophyte and wild relative of rice, is grown normally in saline water. MicroRNAs (miRNAs) are non- coding RNAs that play pivotal roles in every domain of life including stress response. There are very few reports on the discovery of salt-responsive miRNAs from halophytes. In this study, two small RNA libraries, one each from the control and salt-treated (450 mM NaCl for 24 h) leaves of O. coarctata were sequenced, which yielded 338 known and 95 novel miRNAs. Additionally, we used publicly available transcriptomics data of O. coarctata which led to the discovery of additional 48 conserved miRNAs along with their pre-miRNA sequences through in silico analysis. In total, 36 known and 7 novel miRNAs were up-regulated whereas 12 known and 7 novel miRNAs were down-regulated under salinity stress. Further, 233 and 154 target genes were predicted for 48 known and 14 novel differentially regulated miRNAs respectively, which with the help of gene ontology analysis, showed several important biological processes that could be involved in salinity tolerance. Relative expression trends of majority of the miRNAs as detected by real time-PCR as well as predicted by Illumina sequencing were found to be coherent. Additionally, expression of most of the target genes was negatively correlated with their corresponding miRNAs. Thus, the present study provides an account of miRNA-target networking that is involved in salinity adaption of O. coarctata plant.
PC193 (IPPC0650) Genome-wide transcriptome orchestra and involvement of Aux/IAA gene family in chickpea flower development Vikash Singh*, Rohini Garg and Mukesh Jain Functional Genomics, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Flower development is one of the key developmental processes that controls seed setting in angiosperms. However, little is known about the molecular mechanisms regulating flower development in legumes. Employing RNA-seq for 11 tissues/organs/developmental, we identified a large number of differentially expressed genes during flower development in chickpea. Further, we identified several genes expressed exclusively at a particular flower development stage. Various transcription factor families and metabolic pathways involved in flower development were also deciphered. The members of MADS-box family were highly enriched among the transcription factor encoding genes up-regulated during various stages of flower development. These data provide a resource for exploring the complex molecular mechanisms underlying flower development and identification of gene targets for functional genomics in legumes. Further, a genome- wide analysis of Aux/IAA gene family members in legumes identified several members, which expressed specifically at various stages of flower development. Notably, CaIAA2, CaIAA9 and CaIAA13 revealed distinctively higher transcript accumulation during stages of flower development, suggesting their possible role in flower development. Overall, these results would help in investigation of precise functions of Aux/IAA genes in legumes during flower development in chickpea.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 212 Session C : Genomics, Epigenomics and Regulation Biology
PC194 (IPPC0662) Preparation and characterization of resource materials for the isolation of Mediator complex from rice Subhasis Samanta* and Jitendra K. Thakur National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India *Presenting author: [email protected]
Mediator complex acts as a molecular bridge transmitting the regulatory signal from the promoter-bound transcription factor to the RNA polymerase II in the process of transcription. Genetic and molecular analyses have unravelled important regulatory roles of Mediator subunits at every stage of plant life cycle starting from flowering to embryo and organ development, to even size determination. The present study is regarding preparation and characterization of resource materials for the isolation of Mediator complex from rice. We are exploiting both the immuneprecipitation and the ‘Tag’- mediated purification principles for the isolation of rice Mediator complex. Towards the end, we have tested and validated quite a few antibodies against rice Mediator subunits, which help us to detect the chromatographic fractions containing the Mediator complex. Further, the Mediator enriched chromatographic fractions is being used to selectively precipitate the Mediator complex. On the other hand, we have raised rice transgenic lines over-expressing ‘HaloTagged’ OsMED7 and OsMED17, two important subunits of the Mediator complex. The tagged subunit will act like a bait protein to fish-out the entire complex from a sea of thousands of other proteins. We have performed pull-down experiment with the HaloLink resin, which covalently binds with the HaloTagged Mediator subunits like Halo-MED7 and Halo-MED17 of the transgenic calli. The presence and the purity of the isolated Mediator complex are being analysed. The isolated Mediator complex would be useful to understand the structural and mechanistic details of the Mediator functioning in the transcriptional process.
PC195 (IPPC0685) Early axis growth during germination is gravisensitive and mediated by ROS and calcium Rup Kumar Kar* and Khangembam Singh Plant Physiology and Biochemistry Laboratory, Department of Botany, Visva Bharati Univ., Santiniketan, Dist. Birbhum-731235, India *Presenting author: [email protected]
ROS has been explored for their possible role in gravity-induced early axis growth during germination of Vigna radiata seeds. Axis of seeds incubated at any position tends to aligned itself along gravity this resulted in curvature growth due to cell elongation at the convex side (upper side) as observed histologically. Such growth was accompanied with ROS production, visible on the convex side besides accumulation in the apical region of axis, as observed through NBT stain •- for O2 . The accumulation was inhibited completely by ZnCl2 while DPI inhibited accumulation only in the apical region. •- Apart from NADPH oxidase, another O2 producing enzyme, NADH oxidase was revealed during spectrophotometric as well as in gel native PAGE assay. The enzyme was insensitive to DPI and SHAM while inhibited by ZnCl2 as observed through spectrophotrometric assay. Treatment with IAA inhibited early axis growth (predominantly occurring in the radicle) as well as ROS accumulation while TIBA, auxin transport inhibitor, inhibited only curvature growth and ROS •- accumulation at the convex side without affecting elongation growth and O2 accumulation in the elongation zone. Thus gravitropic growth resulting from unequal cell elongation at the upper and lower sides of the axis was a result of spatial regulation of ROS production on the opposing sides of the germinating axis. NADH and NADPH oxidases are the main sources of ROS and the inhibition of NADH oxidase by auxin on the lower side resulted in curvature growth. Pharmacological studies suggest the possible mediation of cytosolic Ca2+ in curvature growth via ROS.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 213 Session C : Genomics, Epigenomics and Regulation Biology
PC196 (IPPC0696) Novel feed-forward loop between two putative plasma membrane enzymes: NADPH oxidase and H+- ATPase Arkajo Majumdar* and Rup Kumar Kar Plant Physiology and Biochemistry Laboratory, Department of Botany, Visva Bharati Univ., Santiniketan, Dist. Birbhum-731235, India *Presenting author: [email protected]
Plants’ growth, development and defence rely largely on the activities of Plasma Membrane (PM) located NADPH oxidase (NOX) and H+-ATPase enzymes whose potential synchronization has been investigated in the roots of two days grown Vigna radiata (L.) Wilczek seedlings. Treatments with CCCP (protonophore) and ortho-Vanadate (P-type ATPase inhibitor) revealed that proton gradient across plasma membrane, generated due to its regulated extrusion by PM H+-ATPase, is crucial for NOX-dependent ROS metabolism. While intensity of NBT staining decreased in histolacalization studies, spectrophotometric (XTT) assays of both intact and extract samples showed lower superoxide production than control and analyses of in-gel Native PAGE assays confirm that NOX-activity of the seedlings grown in either CCCP or ortho-Vanadate decreases. On the other hand, incubation in different ROS scavengers (KI and CuCl2) and NOX inhibitor (ZnCl2) lessened PM H+-ATPase activity in spectrophotometric as well as in Native PAGE assays with the effect of CuCl2 being most pronounced. Interestingly, PM H+-ATPase was more susceptible to CuCl2 than CCCP. Activation of NOX and PM H+-ATPase appears to be positively regulated by [Ca+2]cyt from apoplastic source, since +2 +2 LaCl3 (PM Ca -channel blocker) and EGTA (Ca chelator) repressed both the enzymes. Ethylene, probably by stimulating hyperpolarization-activated Ca+2-channels (HACC), induced the activity of both NOX and PM H+-ATPase whereas,
AgNO3 (ethylene action inhibitor) and CoCl2 (ethylene synthesis inhibitor) inhibited both the enzymes providing additional support to the supposition that NOX and PM H+-ATPase are interdependent.
PC197 (IPPC0698) Characterizing novel miRNA regulating cellulose synthase genes in Arabidopsis thaliana so as to increase the cell wall biomass and bioenergy production Chandrashekhar P. Joshi and Yogesh Ahlawat* Department of Biological Sciences, Michigan Technological University, Houghton, Michigan, USA *Presenting author: [email protected]
Plant cell wall, the formidable, dynamic and complex barrier is governed by more than 1500 genes. Besides providing define structure plant cell wall plays vital role in transport of solvent, solute, intercellular communication and acts as a first immune layer against various pathogens. Plant cell wall constitutes majorly of cellulose with small amounts of hemicellulose, lignin and pectin. Cellulose is synthesized by cellulose synthase complex, rosette terminal domain embedded in plasma membrane. Different forms of cellulose synthases (CesAs) are responsible for the production of cellulose in primary and secondary cell wall. So far, 10 CesA genes in Arabidopsis thaliana have been characterized, which plays an important role in cellulose synthesis. Several attempts have been made to understand the regulation of CesA genes by tiny non coding RNA based regulators called miRNAs. miRNA regulating the expression of cellulose synthases is of intensive research because of their prominent applied consequences like bioenergy production. In the present study using in silico approaches we find out several miRNA which could influence the expression of CesA. Furthermore, we define a network of in silcio software and manual algorithm which could efficiently pull out biologically relevant miRNA out of a pool of several miRNAs. These selected miRNAs can be further validated by several routine molecular biology assays.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 214 Session C : Genomics, Epigenomics and Regulation Biology
PC198 (IPPC0702) Investigations on microRNA-mediated regulatory networks during heat stress in tomato Sombir Rao*, Asosii Paul, Sarita Jha and Saloni Mathur National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Heat stress is a major cause of crop loss worldwide, reducing average yields by more than 50%. Plants have evolved multidimensional regulatory mechanisms to minimize cell damage and maintain cellular homeostasis during elevated temperatures. This hyper-thermal stress is challenged jointly by two major classes of heat stress response (HSR) proteins: the heat shock proteins (HSPs), molecular chaperones and heat stress transcription factors (HSFs) that regulate HSP transcription. Knowledge of the interplay of these HSR components and microRNAs (miRNAs), major class of small non-coding regulatory molecules, in regulating heat stress response is very rudimentary. Till date, only a handful of heat- responsive miRNAs have been reported in plants, none of which have been functionally validated to target any HSF or HSP. Moreover, information on role of HSFs/HSPs in miRNA regulation is also limited. We have identified HSF and HSP members through rigorous motif-based searches in tomato and performed extensive expression analysis of candidate genes in response to different heat stress regimes as well as under other abiotic stress conditions, phytohormones and various plant developmental stages. Conserved as well as novel heat responsive miRNAs have been identified from NGS data generated from control and heat-stressed tomato leaves. Differentially expressed miRNAs targeting HSFs and HSPs as well as regulation of heat-responsive miRNAs themselves by HSFs has revealed a complex regulatory mechanism involving interplay between miRNAs and HSR genes. Our findings have enhanced the understanding of various miRNA-mediated regulatory networks operating in plants to provide thermo-tolerance, leading to applications in crop improvement.
PC199 (IPPC0704) Development of genome wide PCR based InDel markers in chickpea Ankit Jain*, Manish Roorkiwal, Sandip Kale, Mahendar Thudi, Samineni Srinivasan, Pooran M Gaur and Rajeev K Varshney International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India *Presenting author: [email protected]
Chickpea (Cicer arietinum) is the second most important food legume globally, which provides nutritional food security to global population. With the availability of large scale sequencing and re-sequencing data, use of sequence based Insertion-deletions (InDel) markers are gaining popularity because of their genome wide abundance. In addition, InDels are also co-dominant in nature, inexpensive, highly polymorphic and high precision sequence specific PCR based markers. With the objective to extend the marker repertoire and utilization in molecular breeding, re-sequencing data on parental lines of chickpea inter- and intra-specific populations were aligned with reference genome and then Dindel program was used for identification of InDels. As a result 8,307 InDels with >20 bp size among different parental lines were selected. Average distribution of InDels was found 1,038 InDels/LG with maximum number of InDels on LG04 (1952 InDels) and minimum number of InDels on LG08 (360 InDels). To validate and convert these InDels in to PCR based markers, primers were designed for 428 InDels across genome using various criteria. These primers were tested for amplification and polymorphism on the same parental lines. So far we have tested ~200 primers on these parental lines resulting in higher amplification success rate (>90%) with almost 50% polymorphism rate across parents on 3% agarose gel. These gel based markers are expected to contribute and accelerate the genetic studies and support the molecular breeding activities in chickpea.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 215 Session C : Genomics, Epigenomics and Regulation Biology
PC200 (IPPC0706) Identification, characterization and expression profiling of the regulators of small RNA biogenesis and function in response to biotic stresses in select legumes Vanika Garg*, Gaurav Agarwal, Aamir W Khan, Himabindu Kudapa, Lekha Pazhamala, Dadakhalandar Doddamani and Rajeev K. Varshney International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India *Presenting author: [email protected] Small RNAs have emerged as one of the most versatile performers in regulation of gene expression in plants. The biogenesis and functioning of small RNAs is regulated by major proteins like Dicer-like (DCL), RNA dependent RNA polymerase (RDR) and Argonaute (AGO). It is therefore imperative to investigate the role of protein/gene families involved in regulation of expression of small RNAs in important legumes like chickpea, pigeon pea and peanut. HMM, blastp searches, followed by domain scanning identified 4 DCL, 5 RDR, 13 AGO in chickpea; 4 DCL, 5 RDR, 13 AGO in pigeon pea and 7 DCL, 10 RDR, 22 AGO proteins in peanut. Phylogenetic analysis resulted in clustering of these members into groups based on their orthologs from Arabidopsis and soybean. Subsequently, motif analysis, chromosomal distribution and gene structure analysis of these families in the above mentioned legumes were carried out. These results are in concurrence with the previous reports in other species indicating the conservation of small RNA function and structure in plants. Along with conservation, some variations were also observed across the members of the same family in the legumes studied. The promoter analysis of the genes revealed the presence of stress responsive and hormone related cis-acting elements. The identified genes are being validated using qRT-PCR for their role in response to biotic stresses. In summary, this study provides genomic information of the gene families involved in regulation of gene expression and identification of probable candidate genes imparting resistance against stresses in these legumes.
PC201 (IPPC0723) Comparative transcriptome analysis of differentially expressed genes in Pearl millet Pennisetum glaucum (L.) R. Br. during drought stress condition Rajendra Prasad Meena1, Kishor Gaikwad1, Harinder Vishwakarma1, A.R. Rao2, Sarika Sahu2, C. Tara Satyavathi3, Madan Pal4 and Jasdeep Chatrath Padaria1 1National Research Centre on Plant Biotechnology, 2IASRI, 3Division of Genetics, 4Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected] The rapidly changing global climate is leading to enhanced environmental stress conditions which are causing adverse impacts on agriculture and food security. Drought is one of the main environmental constraints to agricultural productivity worldwide. Drought tolerance is the interactive result of different morphological, physiological and biochemical traits. Pearl (Pennisetum glaucum) is most widely grown millet, which has an efficient adaptive mechanism to cope with drought and other environmental stresses. In this study P. glaucum cv. 841B drought tolerant and 411B drought susceptible were subjected to water withdrawal for 10 days, 15 days at post flowering stage. Total RNA was isolated and RNAseq was performed using Roche454. A total of 45 GB raw data was obtained (having 1273008 reads), after trimming of adaptor and unnecessary sequences (showed 1272852). High quality sequences reads were assembled de novo using CLC genomics (average read length of 315.78bp). Unigenes were identified using RPKM analysis and further categorized as biological process, cellular component and molecular function. Total 12883 contigs were obtained and 43% transcripts were annotated using GO database AmiGO2. Around 4361 (78%) of 5583 (~43% of total contigs) had GO category and 585 common transcripts were obtained. Genes were assigned to different metabolic pathways using Expath software. For finding the SSR markers MISA software tool was used and a total of 286 and 153 SSRs were found in drought susceptible and drought tolerant cv respectively. Outcome of this study would be helpful in gene exploration involved in abiotic stress tolerance.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 216 Session C : Genomics, Epigenomics and Regulation Biology
PC202 (IPPC0734) Next-generation sequencing for identification of candidate genes for Fusarium wilt and sterility mosaic disease in pigeon pea (Cajanus cajan) Vikas K. Singh1*, Aamir Khan1, Rachit Saxena1, Vinay Kumar1, Sandip Kale1, Pallavi Sinha1, Annapurna Chitikineni1, Lekha Pazhamala1, Vanika Garg1, Mamta Sharma1, C. Sameer1, Swathi Parupalli1, V. Suryanarayana1, Suyash Patil1, S. Muniswamy2, Anuradha Ghanta3, K.N. Yamini3, P.S. Dharmaraj2 and Rajeev Varshney1 1International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India, 2Agricultural Research Station (ARS)-Gulbarga, University of Agricultural Sciences (UAS), Raichur, Karnataka, India 3Institute of Biotechnology, Professor Jayshankar Telangana State Agricultural University (PJTSAU), Telangana, India *Presenting author: [email protected]
To map resistance genes for Fusarium wilt (FW) and sterility mosaic disease (SMD) in pigeon pea, sequencing-based bulked segregant analysis (Seq-BSA) was used. Resistant (R) and susceptible (S) bulks from the extreme recombinant inbred lines of ICPL 20096 and ICPL 332 were sequenced. Subsequently, SNP index was calculated between R- and S-bulks with the help of draft genome sequence and reference-guided assembly of ICPL 20096 (resistant parent). Seq- BSA has provided seven candidate SNPs for FW and SMD resistance in pigeon pea. In parallel, four additional genotypes were re-sequenced and their combined analysis with R- and S-bulks provided a total of 8362 non-synonymous (ns) SNPs. Of the 8362 nsSNPs, 60 were found within the 2-Mb flanking regions of seven candidate SNPs identified through Seq-BSA. Haplotype analysis narrowed down to eight nsSNPs in seven genes. These eight nsSNPs were further validated by re-sequencing 11 genotypes that are resistant and susceptible to FW and SMD. This analysis revealed association of four candidate nsSNPs in four genes with FW resistance and four candidate nsSNPs in three genes with SMD resistance. Further, in-silico protein analysis and expression profiling identified two most promising candidate genes namely C.cajan_01839 for SMD resistance and C.cajan_03203 for FW resistance. Identified candidate genomic regions/ SNPs will be useful for genomics-assisted breeding in pigeon pea.
PC203 (IPPC0749) Transcriptome profiling of developmental stages of rice varieties varying in carotenoid content Upasna Chettry* and Nikhil Kumar Chrungoo Department of Botany, North Eastern Hill University, Shillong-793022, Meghalaya, India *Presenting author: [email protected]
Carotenoid level is an important quality trait in rice. We have investigated the relationship between carotenoid accumulation and expression levels of genes involved in carotenoid biosynthesis during grain filling in selected cultivars of rice from North East India varying in the bran colour. The grains with coloured pericarp had higher abundance of psy, pds, lcy and βch transcripts. The highest level of expression of these genes was noted in the purple cultivar which accumulated 44.3 μg g-1 of β-carotene, 25.07 μg g-1 of zeaxanthine and 18 μg g-1 of lutein. The carotenoid profile showed a change from β-carotene accumulation to β-carotenoid during maturation. The brown cultivars showed no detectable level of beta carotene, although they had 21 μg g-1 of zeaxanthine and 36 μg g-1 of lutein. Transcriptome profiles generated from two stages of grain filling identified 5812 differentially expressed genes (DEGs) (log2Ratio >1, FDR <0.001) with 1563 transcripts being upregulated while 4249 transcripts were downregulated. While 22% of DEGs could be identified with biosynthesis of secondary metabolites, 15% were identified with biotic stress and 14% with pathways involving signal transduction and starch/sugar metabolism. An insight in carotenoid biosynthetic genes revealed 14 transcripts were upregulated while 5 transcripts were downregulated. Of the upregulated transcripts include psy3, zcd, lcy, zep, cyp4f3 which are related to abiotic stress. Compared to the purple variety, the variety of rice with off white bran showed significant downregulation of psy,1, pds, lcy dhrs4 and zep transcripts during transition from milking to mature stage.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 217 Session C : Genomics, Epigenomics and Regulation Biology
PC204 (IPPC0751) Structure, expression and evolution of dioscorin (alpha Carbonic Anhydrase) in Dioscoreaceae Paramananda Barman* and R. Geeta Department of Botany, University of Delhi, Delhi-110007, India *Presenting author: [email protected]
Dioscorin (an alpha Carbonic Anhydrase; CA) is the major storage protein in the Dioscorea tubers. Dioscorin, a multifunctional protein, consists of two types, dioscorins A and B, with 70% sequence similarity. Here, we present the gene structure, expression and evolution of dioscorin. Genomic sequences of dioscorin A and B, with and without introns, were characterized from Dioscorea and Tacca (Dioscoreaceae). Expression of dioscorin A and B genes in tuber, shoot and leaf was detected at different developmental stages in three cultivars of D. alata (tuberous) and in medicinal D. deltoidea (rhizomatous): a first report for aerial tissues and rhizome. Semi-quantitative analysis revealed that levels of expression of all three paralogs (A1, A2 and B1) were high in tubers in comparison to leaves and shoot tips. Expression was higher in newly emerging tubers compared to the parent tubers (setts). Phylogenetic analyses of dioscorin and related sequences across eukaryotes showed that in Dioscorea, dioscorins formed a strongly supported clade that is distinct from a second group of alpha CAs which are more similar to CAs in grasses. The analyses revealed ancient duplications and independent rapid diversification of alpha CAs across eukaryotes. In plants, duplications occurred in the vascular plant ancestor, before the monocot-eudicot splitting event and within major angiosperm lineages, e.g., monocots and eudicots. Despite extensive duplication and diversification of this apparently fast evolving gene, active sites and secondary structure are highly conserved, suggesting conserved function.
PC205 (IPPC0752) The rice F-box protein, OsFBK, is a component of a novel SCF E3 ligase and is involved in the regulation of anther wall thickenings Pratikshya Borah*, Shane W. Rydquist and Jitendra P. Khurana Interdisciplinary Centre for Plant Genomics, Department of Plant Molecular Biology, UDSC, New Delhi-110021, India *Presenting author: [email protected]
The Ubiquitin-26S proteasome system (UPS) is a large group of proteins involved in the turnover of proteins no longer necessary in the plant system. One of the E3 ligases involved in this process is known as the SCF complex (Skp1-Cullin- Rbx1-F-box) that is instrumental in substrate identification and ubiquitination. The substrate adaptor, F-box, in one of the largest group of proteins found in plants. The present research deals with a rice F-box protein, OsFBK that has a Kelch domain at its C-terminal, a combination unique only to plants. Y2H analysis showed that OsFBK is a part of a novel SCF complex. Expression analysis of this gene reveals its up-regulation in the reproductive stages and root tissues of the rice plant and, also in response to auxin and ABA. Rice transgenics over-expressing OsFBK and its RNAi plants clearly reveal the role of this gene in anther and root development. Electron and confocal microscopy of the transgenic anthers demonstrated differences in their morphology and lignification status. The putative interacting partners of OsFBK were identified from Y2H library of anther tissue and their interactions were validated by Bimolecular Fluorescence Complementation and Co-immunoprecipitation assays. Thus, it could be hypothesized that OsFBK might be involved in lignification processes in anthers, and in controlling proliferation of roots.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 218 Session C : Genomics, Epigenomics and Regulation Biology
PC206 (IPPC0758) Unraveling the genome content of short arm of chromosome 2A of wheat using BAC end sequencing Chanderkant Chaudhary*, Jitendra P. Khurana and Paramjit Khurana Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi-110021, India *Presenting author: [email protected]
Wheat (Triticum aestivum) is an allohexaploid by genetic constitution. It has one of the largest genome of 1700 Mbp which is approximately 40-fold larger than rice. As part of International Consortium on Wheat Genome Sequencing, India is responsible for sequencing of chromosome 2A BAC ends of wheat. Work on sequencing in India started in 2011 with two BAC libraries obtained from Institute of Experimental Botany (Czech Republic) for chromosome 2A of wheat, Triticum aestivum cv. Chinese Spring. So far sequencing of 48,504 out of 55,648 BAC clones has been completed from both the ends generating approximately 97,008 BES sequences. In total, nearly 53.3 Mbp of wheat genomic sequence having an average read length of ~550bp per BAC end has been generated. These BES were mined for repeat sequences using REPEATMASKER revealing approximately 81.9% repetitive elements. Thereafter these sequences were subject to BLAST analyses with different Databases with appropriate E-value (range 1E-10 - 1E-100) and Bit Score (>150). Blast analysis lead to the conclusion that a sizeable number among the significant hits obtained represent the genic region but a vast majority of them represent repeat elements. Protein coding genes which are represented in blast hit are from the diverse biological processes ranging metabolic pathways to signaling pathways. In addition to the above analysis, the BES generated data will help in physical and genetic mapping of wheat genome.
PC207 (IPPC0759) Genome-wide survey and expression analysis of genes involved in RFOs synthesis during development and thermal stress in wheat Naveen Sharma1*, Paramjit Khurana1 1Department of Plant Molecular Biology, University of Delhi, New Delhi-110021, India *Presenting author: [email protected]
The raffinose family oligosaccharides (RFOs) are synthesized by addition of alpha 1,6 galactosyl to sucrose. Increase in levels of RFOs has been associated with the thermal stress such as heat and cold stress. The availability of wheat survey sequence and transcriptome data open avenues to comprehend the genesinvolved in RFOs synthesis during thermal stress. By searching available survey sequence and flcDNA sequences from IWGSC, we identified genes involved in synthesis of RFOs eg. Galactinol Synthase (GolS; EC 2.4.1.82), Raffinose Synthase (RS; EC 2.4.1.82), Stachyose Synthase (STS; EC 2.4.1.67), and Sucrose Synthase (SUS; EC 2.4.1.13). Extensive sequence analysis has been done of these putative RFOs encoding genes to elucidate their structure, chromosomal location and Phylogenetic relationships.Their expression profiling by RNA Seq data analysis, revealedRFOs encoding genes showing specific and co-expression at different stages of wheat plant and in response to thermal stress. The expression analysis of differentially expressing genes were complimented by Real Time PCR analysis. This study implicates the role of RFOs during thermal stress in higher plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 219 Session C : Genomics, Epigenomics and Regulation Biology
PC208 (IPPC0765) Identification of long noncoding RNAs (lncRNAs) in different Capsicum species Vineet Sharma1*, Mukesh Nitin1, Vijaya Bramha2, Sushil Chhapekar1, Toshi Mishra3, Rashmi Gaur1, Sarpras M1, Andrew W. Lynn2 and Nirala Ramchiary1 1School of Life Sciences, 2School of Computational & Integrative Sciences, Jawaharlal Nehru University, New Delhi-110067, India 3Department of Bioinformatics, Banaras Hindu University, Varanasi-221005, India *Presenting author: [email protected] Majority of the eukaryotic genome, around 90% is transcribed into noncoding RNAs, only 1-2% is protein coding. These noncoding RNAs constitutes of long noncoding RNAs (lncRNAs) and small noncoding RNAs (sncRNAs) which performs diverse functions which includes transcriptional interference, chromatin remodelling and histone modifications, alternative splicing and post-transcriptional gene regulation. It indicates that the noncoding region of the genome which was considered to be “junk”, no longer stands true. Recently, Capsicum whole genome has been sequenced which constitutes approximately 80% of transposable and repetitive elements. In this study, our aim was to identify lncRNAs which regulate fruit shape, fruit size, metabolites and develop lncRNA based molecular markers for capsicum breeding program. For this, two Capsicum species with contrasting fruit characteristics and metabolite contents were selected for transcriptome sequencing using Illumina High Seq 2000 and samples were collected from three different fruit stages (Unripe, breaker and ripe), leaf and flower. The de novo assembly of the transcriptome sequences was done by using trinity. The entire dataset from each species after assembly was annotated and protein-coding transcripts were differentiated from noncoding transcripts by using Coding Potential Calculator (CPC). Out of the total transcripts (184975 and 179780), around 85% are noncoding in nature in both species. Some of the differentially expressed lncRNAs are being validated by RT-PCR. Our data reveals that lncRNAs are differentially expressed in tissue and species specific manner and their functional roles creating phenotypic and metabolite diversity between two capsicum species has to be further studied.
PC209 (IPPC0839) Partial sequencing and characterization of enzymes involved in galactomannan biosynthesis pathway in Cyamopsis tetragonoloba L. Monika Verma*, Promila Sheoran, Anoop Singh, Sweta Soni, Archana Yadav, Chhavi Bhardwaj, Kiran Bala, Priyanka Khurana, Sonam Yadav, Usha Kantiwal, Mohit Yadav, Divya Tikaniya and Ashok Chaudhury Dept. of Bio & Nano Technology, Bio & Nano Technology Centre, Guru Jambheshwar University of Science & Tech., Hisar-125001, India *Presenting author: [email protected] Cluster bean, known as Guar, is a large seeded drought-tolerant annual sub-tropical legume of Leguminosae family. Guar gum or its derivatives have tremendous applications in pharma, textile, paper, petroleum, mining, explosives and food industries. Galactomannan biosynthesis pathway begins with sucrose. Cleavage of O-glycosidic bond between glucose and fructose units of sucrose is catalyzed by invertase and sucrose synthase. Sucrose synthase is a glycosyl transferase, converting sucrose in presence of UDP into UDP-glucose and fructose. Direct precursors for galactomannan biosynthesis, GDP-D-mannose and UDP-D-galactose, are formed by actions of GDP mannose phosphorylase and UDP-galactose 4-epimerase. Phosphomannose isomerase converts fructose-6-phosphate (Fru-6-P) to mannose-6- phosphate (Man-6-P). In present investigation RNA was isolated from HG365 and HG870 followed by c-DNA synthesis. Specific and degenerate primers were designed for sucrose synthase, UDP-galactose-4-epimerase, β-mannan synthase, phosphomannose isomerase, α-galactosidase and hexokinase enzymes involved in galactomannan biosynthesis. Conditions were optimized for PCR amplification using cDNA as template by employing specific and degenerate primers. Specific amplified products were sequenced using enzyme specific forward and reverse primers. Sequences were analyzed using BLAST. High degree of similarity was observed with closely related plant species. Alignment data showed a high score of >=200. Phylogenetic analysis of derived sequences and pre-existing sequences showed considerable relatedness among them. This signifies a high level of similarity between the derived sequence and sequences of the enzymes in closely related plants. Attempts are also underway for identification and characterization of major enzymes involved in galactomannan biosynthesis through transcriptome and whole genome sequencing approach in cluster bean.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 220 Session C : Genomics, Epigenomics and Regulation Biology
PC210 (IPPC0842) Identification and modulation of chickpea WRKYs in development and stress Vikas Srivastava*, Kamal Kumar, Savithri Purayannur and Praveen Verma Plant Immunity Laboratory, National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India *Presenting author: [email protected]
The WRKY proteins are one of the promising transcription factors with significance over biological process, metabolism and plant defence. The recent investigation of this TF in many genome calls for further functional studies. This family can be demarcated into group I, II and III WRKYs. The group II have further categorization i.e., IIa, IIb, IIc, IId and IIe. Our analysis indicated 79 WRKY genes in chickpea genome. Among them, CaWRKY5, CaWRKY38, CaWRKY39, CaWRKY44 and CaWRKY61 represents group IIA. Group IIa is smallest clade and have only three representations in Arabidopsis thaliana known for negative regulation of plant defence. Moreover, in Glycine max, 8 members signify WRKY-IIA. Their expression in different developmental tissues suggested mostly ubiquitous expression, with dominance in stem or leaves. Interestingly, CaWRKY39 was found to express in floral tissues. To assess the potentiality of them in plant defence, their response under hormonal (Salicylic acid, Jasmonic acid and Abscissic acid) and pathogenic (Ascochyta rabiei) stresses were investigated. Under such conditions, these were found to up-regulate during SA and ABA treatments; however mixed response were achieved after JA treatment /Ascochyta rabiei infection. Addition to this, these proteins were known to interact together and form homo and hetero-dimer. To look into this possibility of CaWRKY-IIA proteins, Y2H experiments were carried out. The growth on QDO and positive X-gal agarose overlay assay suggested that this feature is well represented in chickpea. Together our experiment suggests the regulation of CaWRKY-IIa during various environmental cues and probably their function may mediate through protein-protein interaction.
PC211 (IPPC0866) Genome-wide transcriptome profiling to unravel mechanism of growth in a sub-tropical bamboo Abhishek Bhandawat*, Gagandeep Singh and Ram Kumar Sharma Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061, H.P., India *Presenting author: [email protected]
Bamboo is one of the most important forest resources of the world. They are widely distributed in tropical, subtropical and temperate forests. It comprises of almost 1200 species worldwide, of which, 130 species representing 18 genera are found in India, making it a major hot spot for bamboo genetic resources. Plant growth is a complex trait determined by various exogenous and endogenous factors. Earlier, transcriptome studies have been carried out in temperate bamboo to identify genes involved in growth. However, molecular basis of growth in tropical/sub-tropical species is unknown. Dendrocalamus hamiltonii (DH) is a commercially important fast growing sub-tropical bamboo. We performed transcriptome profiling of DH to identify the gene(s) and pathways associated with growth. In total, 56.9 million clean filtered reads were generated which were assembled into 39,603 contigs. Of these, 29,032 contigs showed significant hits against nr protein database and 10,256 contigs were differentially expressed during various developmental transitions. We found genes related to growth, cell size regulation, carbohydrate metabolic process and cell wall were substantially enriched in growing tissues. The study suggests that environmental cues, cell signaling pathways, transcription factors and phytohormones were found to regulate expression of genes involved in cell division and elongation leading to overall growth of the bamboo.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 221 Session C : Genomics, Epigenomics and Regulation Biology
PC212 (IPPC0872) Chromatin remodeler in support for bacterial growth in plants Shweta Roy*, Priya Gupta and Ashis Nandi School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Plants defend themselves from pathogens with their well elaborated immune system. All plants possess basal defense which is induced upon recognition of evolutionary conserved molecular patterns. In addition, certain plant-pathogen combinations trigger a strong immune response based on the recognition of effector molecules. Function of most effector molecules remains unknown. By analysing pathogen induced Arabidopsis transcriptome profile, we identified a PRC2 member, which named here as POLYCOMB REPRESSOR FOR DISEASE 1 (PRD1). Along with other PRC2 complex subunits, it silences the target genes through repressive histone modification. Pathogenesis-induced expression of PRD1 is associated with enhanced accumulation of H3K4me3. PRD1 over-expression plants are susceptible to Pseudomonas syringae carrying AvrRpt2 (Pst-AvrRpt2) effector, and are defective in SA, JA and ABA signaling. Through tanscriptome analysis we identified RPS2, which provides resistance against AvrRpt2 in gene-for-gene interaction, as a target of PRD1. PRD1 overexpression plants have reduced basal and pathogen induced expression of RPS2. Chromatin immuno-precipitation experiment shows that PRD1 physically binds at RPS2 promoter and suppresses its expression by enhancing accumulation of H3K27me3, a repressive chromatin modification at RPS2 locus. Through screening of Arabidopsis cDNA library by yeast two-hybrid assay, we identified large numbers of PRD1 interacting partners, and one of them is DROUGHT INDUCED 19 (DI19). RPS2 promoter has a binding motif for DI19. Our results suggest that AvrRpt2 enhances expression of PRD1, which along with DI19 and probably with some other unknown partners make a complex and binds to RPS2 promoter and represses its expression, to support bacterial growth in plants.
PC213 (IPPC0903) Source-sink dynamics and QTLs based options to improve sucrose content in sugarcane stalks Amaresh Chandra1*, Indu Verma1, Kriti Roopendra1, Radha Jain1, Ramakant Rai1 and Yong-Bao Pan2 1Indian Institute of Sugarcane Research, Lucknow-226002, U.P., Indian 2USDA-ARS, MSA, Sugarcane Research Laboratory, Houma, LA 70360, USA *Presenting author: [email protected]
Improving sucrose content in sugarcane stalk is the prime objective that can be addressed through breeding as well as source-sink dynamics associated interventions. Since sugarcane cultivars possess >100 chromosomes (2n = 100 - 130), and are genetically complex polyploid and aneuploids, identification of quantitative trait loci (QTLs) associated with sucrose content is considered the preferable option to improve sucrose content through molecular breeding. In this direction, a linkage map was constructed using AFLP, target region amplification polymorphic (TRAP) and SSR markers. This map had a cumulative genome length of 7406.3 cM that included 675 AFLP (69.8%), 90 TRAP (9.3%), and 202 SSR (20.9%) markers. The 202 SSR markers were assigned to 65 CGs and 8 HGs. Based on this map, 24 putative QTLs affecting sucrose content were identified. Five QTLs were unlinked and the other 19 were located on nine CGs within four HGs. Of these QTLs, 11 had an effect in both, plant cane (20.33%) and first stubble (25.68%) crops. While addressing the source-sink dynamics, the signal communication was perturbed by restricting source leaves’ photosynthetic ability and by improving sink strength with the application of plant hormone like GA3. Results indicated differential accumulation of sugars, along with expression of some pivotal genes in stalk, source and sink leaf tissues. Results propose a model and various options to address this important issue in sugarcane genomic research.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 222 Session C : Genomics, Epigenomics and Regulation Biology
PC214 (IPPC0926) Cloning, phylogenetic analysis and expression profiling of Crocus sativus zinc finger transcription factors Aubid Malik* and Nasheeman Ashraf Plant Biotechnology Division, CSIR-Indian Institute of Integrative Medicine, Sanat Nagar, Srinagar- 190005, India *Presenting author: [email protected]
Crocus sativus is the only plant known to produce appreciable quantities of apocarotenoids, like crocin, picrocrocin and safranal. These metabolites are produced in stigma tissue in developmental stage specific manner. Although, the biosynthetic pathway of apocarotenoids in Crocus is pretty well understood, but the mechanisms regulating tissue and developmental stage specific production of these metabolites is still elusive. Characterization of the transcriptome of saffron stigmas is sine quo non for elucidating the molecular basis of flavor, color biogenesis, genomic organization and biology of gynoecium of saffron. In this backdrop, transcriptome-wide survey of Crocus sativus lead to the identification of 173 zinc finger transcription factors. C3H, C2H2 and AN20/AN1 were the most preponderant classes with 42, 24 and 18 members which grouped into 4, 4 and 3 classes respectively. Besides, expression analysis of representative members of the three classes was carried out in different tissues, at different stages and in response to different stresses and phytohormones. Member of C3H showed highest expression in anther, C2H2 in tapel and AN20/AN1 in Stigma. C3H member was induced by salt, C2H2 by cold and AN20/AN1 by drought. C2H2 member was upregulated by auxin and gibberellic acid, while C2H2 by Methyl jasmonate and ABA and AN20/AN1 by auxin and ABA. Since AN20/AN1 showed highest expression in stigma and in anthesis stage commensurate with the apocarotenoid profile of the flower, was selected for further characterisation. Therefore, we envisage probable role of AN20/AN1 in Crocus sativus specific metabolite regulation, which is under further evaluation.
PC215 (IPPC0938) Genome wide identification expression profiling and characterization of Lectin gene super family of mulberry Bushra Saeed* and Paramjit Khurana Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India *Presenting author: [email protected]
Lectins are a diverse group of ubiquitously present, highly specific sugar binding proteins. Members of this large gene family have been assigned broad biological functions from defense to acting as storage proteins. Despite possessing several interesting characteristics, their functions remain essentially undefined. Mulberry known for its medicinal benefits is also a rich source of lectins. Using an exhaustive HMM-based search we identified the lectin gene complement in Morus notabilis with around 197 members. These putative lectin genes were classified into twelve distinct gene families based on the presence of characteristic sugar-binding domains. Members of this superfamily were assigned varied gene ontologies to identify putative functions and determine cellular localizations. Interestingly, characteristic expression patterns were observed across the lectin superfamily in response to a variety of stress conditions. This is suggestive of specialized functions under diverse conditions possibly by linking the specificity of sugar recognition with mediating precise stress responses in plants. A Jacalin family member showing a drought specific expression was further characterized in Arabidopsis. The identification of putative gene family members from the genus Morus developed in this study can find wide applicability in lectin gene identification and characterization. It can also contribute immensely in the understanding of lectins from mulberry with potential medicinal uses.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 223 Session C : Genomics, Epigenomics and Regulation Biology
PC216 (IPPC0952) Co-expression of regulatory genes involved in protein synthesis and protection to improve cellular level tolerance of tobacco plants under abiotic stress Mahesh Patil*, Chandrashekar Reddy, T.G. Prasad and Udayakumar Makarla Department of Crop Physiology, University of Agricultural Science, GKVK, Bangalore-560065, Karnataka, India *Presenting author: [email protected] Plants when experiences stress there will be reduction in metabolic activities of cell leading to reduced growth and development. In this context, improving cellular level tolerance (CLT) to sustain cell metabolic activities has a phenomenal relevance under stress. Among several CLT mechanism protein turnover and stability are important, which are get affected under stress. Among many regulatory checks in protein turnover, translation and protein protection are very important and several studies have showed relevance of these processes under stress. From this context, to maintain protein homeostasis the best approach will be overexpression of genes/transcription factors involved in these processes. Therefore, an attempt has been made to improve protein turnover and stability by overexpressing three regulatory genes i.e. Pg47 (Pennisetum glaucum 47, a RNA helicase), OseIF4E (Oryza sativa translational initiation factor 4E) and PgHSF4 (Pennisetum glaucum Heat shock factor4) involved in protein synthesis and protection in a model system tobacco. Since, aim was to address both protein synthesis and protection, further, an attempt has been to co-express all these genes together in model system tobacco and comparative analysis was also done. Multigene transgenics showed increased plant growth and tolerance to diverse stresses such as salinity, DTT, PEG, CdCl2 and cold stress with improved seedling growth compared to single gene expressing trangenics. Further, multigene transgenics showed significantly higher protein content and lesser MDA level and also enhanced expression of few stress responsive genes. These results clearly demonstrate that co-expression of specific genes involved in translation and protein stability substantially improved stress adaptation of crops.
PC217 (IPPC0966) Comprehensive transcriptome analysis of Crocus sativus for discovery and expression of genes involved in apocarotenoid biosynthesis Shoib Baba, Tabassum Mohiudin, Aubid Hussain, Zahoor Wani, Nazia Abass and Nasheeman Ashraf* Plant Biotechnology Division, CSIR-Indian Institute of Integrative Medicine, Sanat Nagar, Srinagar, India *Presenting author: [email protected] Crocus sativus stigmas form rich source of apocarotenoids like crocin, picrocrocin and safranal, which besides imparting color, flavour and aroma to saffron spice also have tremendous pharmacological properties. Inspite of their importance, the biosynthetic pathway of Crocus apocarotenoids is not fully elucidated. Moreover, the mechanism of their stigma specific accumulation remains unknown. Therefore, deep transcriptome sequencing of Crocus stigma was done to identify the genes and transcriptional regulators involved in the biosynthesis of these compounds. Transcriptome of stigma was sequenced using Illumina Genome Analyzer IIx platform, which generated 51,350,714 stigma reads. Sequences were assembled de novo using trinity resulting in 64,438 transcripts which were classified into 32,204 unigenes comprising of 9,853 clusters and 22,351 singletons. A comprehensive functional annotation and gene ontology (GO) analysis of all the sequences was carried out. Many genes involved in carotenoid/apocarotenoid pathway were identified for the first time in this study which includes zeta-carotene isomerase and desaturase, carotenoid isomerase and lycopene epsilon-cyclase. GO analysis showed that the predominant classes in biological process category include metabolic processes followed by cellular processes and primary metabolic processes. KEGG mapping analysis indicated that pathways involved in ribosome, carbon, starch and sucrose metabolism were highly represented. Differential expression analysis indicated that key carotenoid/apocarotenoid pathway genes including phytoene synthase, phytoene desaturase and carotenoid cleavage dioxygenase 2 are enriched in stigma thereby providing molecular proof for stigma to be the site of apocarotenoid biosynthesis. This data would provide a rich source for understanding the carotenoid/ apocarotenoid metabolism and flower development in C. sativus.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 224 Session D : Plant Metabolism and Metabolomics
PD218 (IPPC0247) Increased glyphosate resistance gene (igrA) confers resistance to the herbicide glyphosate in transgenic indica rice Vijay Sheri*, Bhabesh Borphukan, Mohan Acharya and M.K. Reddy Plant Molecular Biology, International Centre for Genetic Engineering & Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Plants expressing Pseudomonas sp. strain PG2982 increased glyphosate resistance gene (igrA) are known to be resistant to glyphosate, a potent herbicide that inhibits the activity of the endogenous plant EPSPS. In order to develop herbicide- resistant indica rice, the disarmed Agrobacterium strain EHA 105 harbouring the binary vector pMDC-99 was used for transformation. The T-DNA region of the vector carries igrA gene with a DNA segment encoding a chloroplast transit peptide, which would direct the igrA to the organelle where the shikimate pathway is located, under the control of rice actinII promoter for constitutive expression, and hygromycin phosphotransferase gene (hpt) as a plant selectable marker for selection on hygromycin. A large number of transgenic rice plants were obtained with hygromycin and glyphosate, most of the transformants showed fertile. The integration and expression of igrA gene from regenerated plants was analyzed by PCR and Southern blot analysis. Transgenic T1 seeds were germinated on hygromycin and transferred them to glyphosate medium to assess the glyphosate tolerance. The transgenic rice plants had more glyphosate resistance than the wild-type plants. Field test of transgenic rice plants further confirmed that they survived 100% spray of glyphosate (Roundup) at a regular dose used for conventional rice weed control.
PD219 (IPPC0261) Influence of arsenate and phosphate on growth and thiol metabolism in two cultivars of rice (Oryza sativa L.) Jayeeta Saha* and Asok Kumar Biswas Department of Botany, University of Calcutta, Kolkata-700019, West Bengal, India *Presenting author: [email protected]
Arsenic pollution has become a serious problem since the last decade in West Bengal owing to extensive withdrawal of water for irrigation and portable purposes. Arsenic (As), a metalloid, is toxic to all forms of life including plants. There is a competitive interaction between arsenate and phosphate for the same uptake system in plants as phosphate is chemically similar to arsenate, both belonging to group (Va) of the periodic table. The influence of arsenate and phosphate on growth, total glutathione content and activity of its regulatory enzymes viz. GPX, GR and GST were investigated in 21 day old rice (cv. Nayanmoni and cv. IR-64) seedlings. Arsenic toxicity triggered formation of reactive oxygen species. Thus, α the levels of oxidative stress markers viz., proline, H2O2, non-enzymatic antioxidants like glutathione, -tocopherol, ascorbate, including activity of enzymatic antioxidants viz. ascorbate peroxidase, catalase were altered in the test cultivars; the effects being more significant in IR-64 in contrast to cv.Nayanmani. Phytochelatins (PCs), a class of small thiol-rich peptides, can detoxify the excess of toxic heavy metals by forming a metal-PCs complex in which the heavy metal is bound to the thiol group of Cys unit and therefore, can be used as a potential biomarker for evaluating metal toxicity. Different types and levels of PCs were detected in the roots and shoots of the test cultivars grown in arsenate
(Na2HAsO4.7H2O) with or without phosphate (KH2PO4) by RP-HPLC. Therefore, PCn analysis plays an important role in the study of metal toxicity and tolerance mechanism.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 225 Session D : Plant Metabolism and Metabolomics
PD220 (IPPC0262) Physiological, phytochemical and molecular studies on antioxidant accumulation in the medicinal plant Amaranthus spinosus Linn. as influenced by abiotic stress G. Garggi* and Roy Stephen Plant Physiology, Kerala Agricultural University, College of Agriculture, Vellayani, Thiruvananthapuram-695522, Kerala, India *Presenting author: [email protected]
Plants continue to serve mankind as food and medicine since time immemorial. The medicinal effects in plants are due to the presence of various bio-active principles which are the products of secondary metabolism in plants. Plants have evolved the secondary metabolic pathway mainly to defend themselves from numerous biotic and abiotic stresses. These stresses increases production of reactive oxygen species (ROS), that creates oxidative stress to plants. Antioxidants maintain the equilibrium of generation of ROS and its scavenging. It is found that abiotic stress acts as an environmental cue for the enhanced production of antioxidants, thereby increasing the therapeutic value of medicinal plants. A study was conducted to investigate the influence of abiotic stress factors viz. low light stress and water stress, on the growth characteristics, physiological and biochemical parameters as well as on the various enzymatic and non-enzymatic antioxidants in Amaranthus spinosus Linn. It is an annual herb found throughout India and is potent medicinal plant known for its antioxidant accumulation. Growth parameters like plant height and leaf area had a positive response with shade. Starch content was high for the plants under open environments. Enzymatic and non-enzymatic antioxidants in the plant showed variations across different treatment combinations of shade and water stresses. Expression study of the gene chalcone synthase (CHS), the key enzyme in Phenyl Propanoid pathway in the biosynthesis of flavonoids and phenolic compounds, was also done using Reverse Transcriptase-PCR (RT-PCR) and was related with the intensity of stress.
PD221 (IPPC0335) Gas-chromatography mass-spectrometry (GC-MS) based metabolite profiling in two contrasting germplasms of rice in response to salinity stress Khalid Anwar1*, Nita Lakra2, Sneh Lata Singla-Pareek2 and Ashwani Pareek1 1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Salinity is a major factor that limits rice production worldwide. Gas chromatography mass spectrometry (GC-MS) is frequently used to provide the sensitive and reproducible detection of hundreds to thousands of metabolites in a single tissue sample. Untargeted metabolite profiling has the potential to identify numerous novel metabolites. Here, we developed a method for GC-MS based metabolite profiling for rice, in order to gain an overview of the pathway of primary metabolism. In order to reveal the physiological and molecular differences in salt tolerance between wild (Pokkali) and cultivated variety of rice (IR64), profiles of more than 100 of key metabolites were studied using GC-MS. In the present study, untargeted and targeted metabolic profiling analyses were carried out in rice tissues. Principal component analysis (PCA) was employed to explore the metabolite profiles from rice shoot samples at eight different time points. Significant changes in metabolite levels were found in both Pokkali and IR64 in response to salinity with tryptophan, proline and lysine being accumulated at much higher levels in the Pokkali as compared to IR64. In contrast, glycine, homoserine, threonine and alanine levels were higher in IR64 as compared to Pokkali, as IR64 probably adjusted its cellular homeostasis mainly through increasing sugars levels at initial stress exposure. We also found that overall amino acids, sugars were increased whereas, organic acid decreases during salinity stress treatment. A possible role of these metabolites in response to salinity will be discussed.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 226 Session D : Plant Metabolism and Metabolomics
PD222 (IPPC0406) A comparative evaluation of GC-MS based phytochemicals, in vitro radical scavenging properties, antioxidant content and yield performance of two promising accessions of Amaranthus hypochondriacus Aditya Manashi* and Bhattacharjee Soumen Department of Botany, Centre for Advanced Study, University of Burdwan, Burdwan-713104, West Bengal, India *Presenting author: [email protected]
The present study aims at studying two promising accessions of Amaranthus hypochondriacus (IC107144 and IC94661) based on their antioxidant and radical scavenging properties, types of hydroxyl rich phytochemical availability and yield potentials. Quantitative antioxidant profiling of methanolic leaf extracts were done based on in-vitro radical scavenging properties (ABTS, FRAP and DPPH radical scavenging assay) and availability of individual antioxidants (Total polyphenol, monophenol, diphenol, flavonoids, betacyanine, anthocyanin, carotenoids). A comparative evaluation of antioxidants from leaf extract reveals a significantly higher antioxidant availability in accession IC107144 as compared to IC94661. The same trend of result were observed when the antioxidant potential of the methanolic leaf extracts of both the accessions were estimated and compared by FRAP, ABTS and DPPH radical scavenging properties. The data of in-vitro radical scavenging properties of both the accessions exhibited significant correlation between individual antioxidant contents. When GC-MS study was carried out, it exhibited presence of several phytochemicals in both the accessions, some of which were having antioxidant properties. The accession IC107144 exhibited presence of some additional compounds, which contain hydroxyls, responsible for radical scavenging properties. So taken as a whole, the data of quantitative antioxidant profiling and GC-MS based qualitative evaluation of phytochemicals from leaf extracts of two accessions of Amaranthus hypochondriacus not only provide evidence of rich sources of antioxidant, radical scavenging properties and availability of several phytochemicals with hydroxyls but also prove that accession IC107144 as better compared to accession IC94661.
PD223 (IPPC0419) Role of phenylalanine for the production of flavonoids in callus cultures of Prosopis cineraria Druce cultivar K1 and Ziziphus mauritiana cultivars Gola and Seb Ruchi Nag Shrivastva*, Arjit Chaturvedi and T.N. Nag Department of Biotechnology, M.N. Institute of Applied Sciences (MGS University) Bikaner-334022, Rajasthan, India *Presenting author: [email protected]
Plant in vitro cultures are able to produce and accumulate many medicinally valuable secondary metabolites. Flavonoids are an important group of medicinal preventive compounds as well as food additives inhibiting detrimental changes of easily oxidizable nutrients. Many different in vitro approaches have been used for increasing biosynthesis and the accumulation of flavonoid compounds in plant cells. The present study summarized the achievements of in vitro technology for the production of flavonoid compounds by focusing on different approaches to improve the production of these compounds. Unorganized callus tissues of plant species were established on MS (Murashige and Skoog, 1962) medium supplemented with different phytohormones. Callus tissues were transferred to fresh MS medium singly supplemented with various concentrations [25, 50, 75 and 100 mg (100 ml)-1 of media] of DL-b-phenylalanine and allowed to grow up to maximum growth indices. Total flavonoid (kaempferol and quercetin) content showed a marked increase in the tissues fed with phenylalanine and gradual increase in the growth of tissues was observed. Highest amount of flavonoid was observed in tissues grown on MS medium supplemented with 100 mg (100 ml)-1 as compared with other concentrations of phenylalanine in the medium in both the cultivars of Ziziphus mauritiana, viz., Seb (8.45 mg g-1 dry wt.) and Gola (8.29 mg g-1 dry wt.) but in the case of P. cineraria highest concentration of 11.23 mg g-1 dry wt. was obtained with 75 mg phenylalanine (100 ml)-1 medium. Enhancement by feeding phenylalanine supports the pervious findings that this amino acid acts as precursors of flavonoids. The results will be discussed in the light of contemporary data.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 227 Session D : Plant Metabolism and Metabolomics
PD224 (IPPC0420) Antioxidants in arid zone plants and their tissue cultures T.N. Nag* Department of Biotechnology, M.N. Institute of Applied Sciences, MGS University, Bikaner–334022, Rajasthan, India *Presenting author: [email protected]
Antioxidants are an important group of medicinal preventive compounds as well as food additives inhibiting detrimental changes of easily oxidizable nutrients. Many in vitro approaches have been used to increase biosynthesis and the accumulation of antioxidant compounds in plant cells. Antioxidants are capable of stabilizing, or deactivating free radicals before they attack cells, and are critical for maintaining optimal cellular and systemic health and well-being. These are gaining lot of importance as a panacea for a large number of life-style diseases like aging, cancer, diabetes, cardiovascular and other degenerative diseases etc. Thus, investigation of various antioxidants present in arid zone plants and their tissue cultures is the demand of today. Cultures of 21 arid zone plants, established from hypocotyls/seedlings/shoot tips were maintained on MS medium supplemented with different concentration of hormones for 12 months by frequent sub- culturing. In this paper various types of antioxidants identified in plant parts and tissue culture will be discussed in the light of contemporary data. The enhancement of these compounds by addition of different concentration of precursors and phyto-hormones will also be reviewed.
PD225 (IPPC0441) LC-MS based approach for metabolic fingerprinting of Capparis moonii, Wight Nutan Malpathak and Pallavi Yadav* Department of Botany, Savitribai Phule Pune University, Pune-411007, Maharashtra, India *Presenting author: [email protected]
Capparis moonii, Wight is an endemic plant belonging to biodiversity hotspot region of Western ghats with ethnomedicinal importance. Therefore, C. moonii emerges as a potential candidate for Bioprospecting. Sequential polarity wise extraction coupled with sonication was adopted for extraction of secondary metabolites from Capparis moonii. An LC-MS based metabolomics approach was used to characterize secondary metabolite quantitative and qualitatively in stem and leaves of C. moonii. PCA, a multivariate technique was used for the analysis of metabolomics data.Current findings are significant in establishing metabolic fingerprint of an unexplored plant and probably paves way for reverse pharmacognosy approach in drug discovery.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 228 Session D : Plant Metabolism and Metabolomics
PD226 (IPPC0445) Modulation of photorespiratory metabolism by oxidative and photooxidative stress in leaf discs of Pisum sativum Ramesh B. Bapatla* and A.S. Raghavendra Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India *Presenting author: [email protected]
Photorespiration is known to protect plants from photoinhibition and is considered to be an important component of plant adaptation to oxidative stress. Reactive oxygen species (ROS), such as H2O2, are generated under oxidative stress conditions. Menadione, a redox active quinone, interferes with mitochondrial electron transport and generates free radicals, causing oxidative stress. We studied the effect of oxidative stress, induced in mitochondria by menadione on enzymes localized in different cell organelles of pea (Pisum sativum) leaf discs. The enzymes were glycolate oxidae (GO), catalase (CAT) and hydroxypyruvate reductase (HPR), all in peroxisomes, while aconitase was located in mitochondria and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in cytosol. The enzyme activities were examined after incubation in dark, moderate (300 μE m-2 s-1) and high light (1200 μE m-2 s-1) intensities. The activities of GO and CAT were increased by menadione upto 40 to 50%, particularly in high light. In contrast, the levels of HPR increased marginally by <20%. The activity of aconitase was marginally decreased. Our results indicate that oxidative stress condition, induced by menadione, modulates markedly the redox-sensitive key photorespiratory enzymes, particularly in light. The enzymes located in mitochondria (e.g. aconitase) or cytosol (GAPDH) did not respond as strongly as those in peroxisomes. We suggest that oxidative stress, originating in mitochondria can affect photorespiratory components in peroxisomes.
PD227 (IPPC0478) Waterlogging responsive MaAP2 an AP2/ERF protein from Mentha arvensis regulates carbohydrate metabolism in Arabidopsis through bidirectional sugar transporter AtSWEET10 Ujjal J. Phukan1*, Sonal Mishra2 and Rakesh K. Shukla1 1Department of Biotechnology, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow-226015, India 2National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Waterlogging is accountable for huge loss of yield as well as quality in commercial and agronomical breeds. MaAP2 an ERF identified in Mentha arvensis waterlogging subtractive cDNA library was showing induced expression in dehydration, cold, salt and waterlogging treatment. It was also induced in response to ABA and ACC. We observed that it binds specifically to different cis-elements DRE, GCC and Jasmonic acid response element which was competed out with excess of cold probe. We have also mapped a potential transactivation domain in MaAP2 adjacent to the DNA binding domain. MaAP2 over-expressing Arabidopsis lines displayed increased tolerance to dehydration, salt and waterlogging treatment in comparision with WT. Microarray analysis of MaAP2 overexpressing transgenic lines revealed 7889 and 4895 up- and down-regulated transcripts, respectively. As carbohydrate metabolism is deeply affected during waterlogging stress we screened the data for sugar metabolism and transport related genes. AtSWEET10 involved in bi-directional sugar transport was highly up-regulated along with other SWEET and Sugar transporter family proteins. We showed that MaAP2 directly binds to the promoter of AtSWEET10 in in vivo and in vitro conditions, allowing MaAP2 to modulate carbohydrate metabolism which would help the plants to survive under adverse environmental stress condition. The study provides novel information about the role of AP2/ERF protein and their role in regulation of carbohydrate metabolism under adverse environmental condition.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 229 Session D : Plant Metabolism and Metabolomics
PD228 (IPPC0528) Association of galactomannan hydrolyzing enzymes activity with early seed germination in rice Nakul Gupta1, M.B. Arun Kumar1*, Jacob Sherry Rachel2, S. Gopala Krishnan3, Malavika Dadlani1, C. Viswanthan4 and M. Nagarajan5 1Division of Seed Science and Technology, 2Division of Germplasm Conservation, 3Division of Genetics, 4Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India 5ICAR-IARI Regional Center, Aduthurai, India *Presenting author: [email protected]
The scarcity of water and labour is forcing the rice farmers to shift from transplanted method of cultivation to direct- seeded cultivation. Faster germination of seed as well as establishment of vigourous crop is of paramount importance in direct-seeded crop. Rice being an albuminous seed, it is hypothesized that galactomannan hydrolysing enzymes plays important role in loosening of the embryo-surrounding tissue, which in turn governs the speed of germination. To test this hypothesis and the effect of gibberellins and Abscisic acid on enzyme activity, studies were conducted on four rice genotypes comprising of two early germinating (<28 hours) and two late germinating (>46 hours) genotypes. Activities of galactomannan hydrolysing enzymes (β-mannanase, β-mannosidase and α-galactosidase) increased with germination in both early and late germinating rice genotypes. The average enzyme activities of the genotypes was higher in early germinating group in comparison with the late germinating group for all the three enzymes and at all the three stages of germination (dry seed, lemma rupture and radicle emergence). Among the enzymes, activity of β-mannosidase was found higher in dry seeds of both the group followed by α-galactosidase and β-mannanase. GA supplement (50 μM) in the germinating medium has increased the activities of all the three enzymes that manifested in faster lemma rupture and radicle emergence in both early and late germinating rice genotypes by 3-5 hrs and 8-12 hrs, respectively. Supplement of ABA (50 μM) in the germination medium has decreased the activities of all the enzymes that resulted delayed seed germination.
PD229 (IPPC0569) Modulation of photorespiratory metabolism by osmotic stress in leaf discs of pea, Pisum sativum Fareed Khan* and A.S. Raghavendra Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India *Presenting author: [email protected]
Plants need to adapt to stress conditions. It is proposed that photorespiration may be an important component of such plant adaptation to abiotic stress, but there is no direct evidence. We therefore, studied the effect of osmotic stress caused by 1.0 M sorbitol on the photorespiratory enzymes in leaf discs of pea, Pisum sativum. To ensure osmotic stress, the pattern and consequences of reactive oxygen species (ROS) in pea leaf discs were monitored on exposure to 1.0 M -2 -1 -2 -1 sorbitol for 3 h under dark, moderate (300 μE m s ) and high light (1200 μE m s ) intensities. H2O2 and superoxide levels were estimated by using DAB and NBT, respectively. Upon exposure of pea leaf discs to 1.0 M sorbitol and light the levels of H2O2 as well as superoxide were increased significantly. Levels of H2O2 increased by 80 %, while that of superoxide increased by 60%. We then looked at the activities of three photorespiratory enzymes located in peroxisome, namely glycolate oxidase, catalase and hydroxypyruvate reductase in pea leaf discs. There was a marked increase in the activity of glycolate oxidase (43%), followed by catalase (20%) and hydroxypyruvate reductase (14%). Experiments to cross- check and compare the activities of aconitase (mitochondrial enzyme) and glyceraldehyde-3-phosphate dehydrogenase (cytosolic) are in progress. Our data suggest that exposure of pea leaf discs to osmotic stress, particularly under high light, can upregulate key photorespiratory enzymes.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 230 Session D : Plant Metabolism and Metabolomics
PD230 (IPPC0577) GC-MS evaluation of bioactive compounds of whole plant ethanol extract from Osmunda hugeliana C. Presl. Manisha Kale* Department of Botany, Jaysingpur College, Jaysingpur-416101, India *Presenting author: [email protected]
Osmunda hugeliana C. Presl. belonging to family Osmundaceae is one of the medicinally important pteridophyte used as tonic, styptic and also for rickets, rheumatism and for intestinal gripping. In this study, the bioactive compounds of O. hugeliana have been evaluated using Shimadzu make QP-2010 GC-MS. The chemical composition of the whole plant ethanol extract was investigated. While the mass spectra of the compounds found in the extract were matched with the NIST library. The compounds were identified by comparing their retention time and peak area with that of literature and interpretation of mass spectra. GC-MS analysis revealed the presence of sixteen bioactive compounds. The most compounds were 3, 7, 11, 15-tetramethyl-2-hexadecen-1-ol, tetradecanoic acid (myristic acid), n-hexadecanoic acid, hexadecanoic acid ethyl ester and octadecanoic acid. Tetradecanoic acid (myristic acid) is having antioxidative, lubricative, hypercholesterolemic, cancer preventive properties, and also used in cosmetic. N-Hexadecanoic acid (palmitic acid) is also having antioxidantive, hypochloresterolenic, nematisidic, pesticidal, lubricative, antiandrogenic flavor, hemolytic properties.
PD231 (IPPC0602) Stage-specific regulation of triglyceride synthesis during seed development in Jatropha curcas L. Bharatula Chaitanya*, Sumit Kumar, P.S. Sastry and Attipalli R. Reddy Department of Plant Sciences, University of Hyderabad, Hyderabad-500071, India *Presenting author: [email protected]
The present study describes the changes in lipid profiles as well as acyl fluxes occurring in developing seeds of a potential biofuel tree species, Jatropha curcas. Metabolic labeling with 14C-acetate indicated a relationship between phosphatidylcholine (PC) and triacylglycerol (TAG) with seed development (34, 37 and 40 days after anthesis, DAA). The developing endosperm preferred synthesis of phospholipids to neutral lipids at 34 DAA. Study of acyl fluxes showed that PC, diacylglycerol (DAG) and TAG were rich in palmitic acid (16:0). From 37 to 40 DAA, the developed endosperm was actively involved in storage TAG synthesis. Fatty acid methyl ester analysis showed presence of oleic acid (C18:1n-9) rich PC, DAG and TAG molecules. This shift in fatty acid composition of PC molecules (16:0 to 18:1) was concomitant with synthesis of 18:1n-9-rich TAGs during these stages. Furthermore, distinct pools of DAG were observed with turnover of acyl-edited C18-rich DAGs showing higher unsaturation at 40 DAA which was confirmed by analysis of molecular species and positional distribution. The expression of diacylglycerol acyltransferase (DGAT2) was correlated with 16:0- rich TAGs at 34 DAA, whereas expression of DGAT1 was significant during 37 and 40 DAA, which was related to 18:1n- 9-rich TAGs. Our data demonstrate metabolic reprogramming and existence of alternate pathways during TAG synthesis involving distinct pools of PC and DAG, presumed to react with each other and contribute to storage TAG synthesis during seed development in Jatropha.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 231 Session D : Plant Metabolism and Metabolomics
PD232 (IPPC0733) Non-targeted metabolomics of a medicinal plant cumin (Cuminum cyminum L.) shoots under salinity Sonika Pandey*, Manish Kumar Patel, Avinash Mishra and Bhavanath Jha Division of Marine Biotechnology and Ecology, CSIR-CSMCRI, Bhavnagar, Gujarat, India *Presenting author: [email protected]
Spice crops occupy a valuable place in traditional medicinal applications and act as food adjuncts because of distinct aroma and flavour. Cuminum cyminum is an important aromatic herbaceous spice crop belonging to the Apiaceae family, cultivated in semi-arid regions of the world. India is the largest producer, consumer and exporter of cumin seeds and contributes up to 90% of total productivity. The present study is intended to comprehensively analyse the composition of metabolites under salinity that provides the physiological status of a plant to combat stress conditions. Cumin is an active reservoir of metabolites (primary and secondary) or phytochemicals that are characterized as amino acids, flavonoids, polyphenolic and terpenoids etc. Different analytical methods, such as LC MS, GC MS, etc. have been used to elucidate comprehensive information about metabolites. Cumin shoots contain 53% polyunsaturated fatty acids. Saturated FAs increased gradually with salinity, whereas unsaturation index and degree of unsaturation changed arbitrarily along with the percent quantity of unsaturated FAs. A total of 45 differentially expressed metabolites were identified, including luteolin, salvianolic acid, kaempferol and quercetin, which were phenolic, flavonoid or alkaloids in nature and contain antioxidant activities. Additionally, metabolites with bioactivity such as anticancerous (docetaxel) and antimicrobial (megalomicin) were also identified. The study showed that cumin shoots are rich source of metabolites with bioactivities, essential amino acids, phenolic compounds and fatty acids, which unveil its medicinal characteristics, providing useful insights about metabolic responses under salinity that creates its utility in nutraceutical industries.
PD233 (IPPC0736) Metabolomics of medicinal and spice plants: A tool for quality check of herbal products Agepati Raghavendra and Sarada Tetali* Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India *Presenting author: [email protected]
Metabolomics provide a snap shot of biochemical status of an organism, by elucidating the spectrum of metabolites within. Plants are estimated to contain more than 200,000 different metabolites. Such wide diversity of phytochemicals and their occurrence spread across aerial and underground organs makes “plant metabolomics”, a daunting task. However, recent technological advances in metabolomics made the exploration of such treasure easy and discovery of novel natural plant products possible. The combination of liquid/gas chromatography and mass spectrometry provides a common base of metabolomics though there are several other techniques like nuclear magnetic resonance (NMR) in use. Metabolite libraries of Indian medicinal and spice plants have largely remained unexplored. The metabolite fingerprints of medicinal and spice plants not only help in the discovery of drug molecules but also can be exploited for quality check of herbal samples including herbal medicines and spices for commercial use. We have attempted to perform metabolite profiling using GC-MS and/or LC-ESI-MS/MS of several medicinal plants, which are used for treating human inflammatory diseases, e.g. Ocimum sanctum (leaves), Terminalia arjuna (stem-bark), Boswellia serrata (gum resin), Hemidesmus indicus (roots) and of Curcuma longa and C. aromatica (rhizomes). Further, we have tried to compare metabolite variation in rhizomes of seven cultivars of Curcuma. The possible use of metabolomics data to differentiate the cultivars and to assess the quality of turmeric samples would be discussed.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 232 Session D : Plant Metabolism and Metabolomics
PD234 (IPPC0901) CCPM v3.4: towards collaborative metabolomics Indira Ghosh1, Abhijit Mitra2, Nita Parekh2, VikramPudi2, BrotoChakrabarty2, Prashanthi Dharanipragada2, Ramya Gurrapu2, Narendra Babu Karuputhula2, Manoj Kumar Sekhwal1, Gaurav Sharma1*, S.R. Kiran Raj2, Manoj Kumar Maramreddy2, V.P. Srivani2 and V. Dharma Teja2 1School of Computational and Integrative Science, Jawaharlal Nehru University, New JNU Campus, New Delhi-110067, India 2Center for Computational Natural Sciences and Bioinformatics, International Institute of Information Technology Hyderabad, Gachibowli, Hyderabad-500032, India *Presenting author: [email protected]
Assisted by advances in high-throughput instrumentation, metabolomics has emerged as a popular research area, which is witnessing rapid generation of large volumes of high quality data. This demands ready availability of advanced analytical and computational methods for generating analyzed datasets, as well as advanced tools for data analysis, visualization and integration; within a collaborative platform which addresses the unique interdisciplinary character of metabolomics. The Computational Core for Plant Metabolomics, CCPM v3.4, is a fully functional, open, state of art web application, addressing the issues of storage, processing, analysis and sharing of data and results of metabolomics experiments. With a python web framework (web2py) backend, CCPM allows rapid prototyping, easy maintenance and role-based access control without compromising on security, scalability and efficiency. Accessible through a user friendly bootstrap- based GUI, CCPM functions as a secure repository with a pipeline for end-to-end analysis of LC/GC-MS data involving raw data capture, data pre-processing, putative metabolite characterization (linking to METLIN and GOLM), data pretreatment, and a wide range of statistical analysis, with option for customization of parameters from the web interface itself. Bypassing the pipeline, the modular architecture of CCPM also allows inputting of externally preprocessed/pretreated data, directly, to available analysis tools. In contrast with task size restrictions, associated with usual stateless or session- oriented web applications, CCPM supports long-running tasks featuring task completion alerts. Other features include support for handling data in all popular file formats, user friendly tools for on the fly data analysis and visualization, and provision for publication quality image download.
PD235 (IPPC0910) Comparative physiological basis of variability in germination, seedling vigour and vegetative vigour in wheat and barley Madhurima Das*, Rakesh Pandey, Vijay Paul and Attar Singh Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India Presenting author: [email protected]
Wheat and barley are two important cereal crops having similar phenological stages but different growth and developmental behavior. A comparative study on the physiological basis of variability was made in wheat and barley in terms of germination rate, seedling growth, vegetative growth and its relation to cell wall properties (ferulic acid content), assimilate partitioning to roots, root distribution in soil profile. The study showed that the wheat genotype (HD2824) showed a faster germination rate, due to its ability to maintain lower critical water potential of seed, higher β amylase activity, shorter duration of the IInd phage in triphasic imbibition curve than the barley genotype (RD2668). However, the seedling and vegetative vigour was higher in barley than wheat. At 15-21 DAS in field, the seedling vigour of barley was greater as compared to wheat similarly, the vegetative vigour was greater in barley than wheat in relation to higher dry matter production, leaf area, specific leaf area, no of nodal root, root length. The higher specific leaf area was negatively associated with cell wall ferulic acid, which helps in cell wall lignifications, thus limiting cell expansion. Root distribution study was done by trench profile wall method (1 m depth), showed a higher percentage of total root distribution in the upper soil horizon (0-30 cm) in barley as compared to wheat. The present study summarise that wheat genotype showed faster germination rate than barley genotype but the seedling and vegetative vigour was higher in barley as compared to wheat.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 233 Session D : Plant Metabolism and Metabolomics
PD236 (IPPC0970) Interactome analyses in the stress associated signaling cascade: A case for the legume crop chickpea (Cicer arietinum L.) Savithri Purayannur*, Kamal Kumar, Vikas Srivastava, Shreenivas K. Singh and Praveen K. Verma National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected] Plants have evolved many signaling cascades that relay multiple environmental and developmental signals thus compensating their sessile nature, especially in harsh environments. The legume crop chickpea (Cicer arietinum L.) was taken as a system to identify interactions among the components of MAPK signaling cascade and their targets. We identified 7 MAPKKs and 16 MAPKs from the chickpea genome. These gene families were classified into four groups each, based on the model plant Arabidopsis classification. Many alternatively spliced forms of MAPKs were also identified, which increases the signaling repertoire. The MAPKs had 2, 5, 1 and 8 members in groups A, B, C and D respectively. The MAPKKs group A, B, C and D had 3, 1, 1 and 2 members respectively. The open entry clones of 7 MAPKK, 6 MAPKKEE and 21 MAPKs were generated. To identify the interactions by split-ubiquit in based yeast-two hybrid (Y2H) system, we mobilized these ORFs into bait and prey destination vectors. Interaction analysis revealed many unique interactions, some of which were isoform-specific. Our study thus highlighted the importance of alternative splicing in MAPK signaling by direct protein-protein interaction. Selected interactions among these members were confirmed in- planta by bimolecular fluorescence complementation (BiFC) analysis. We also studied the interaction of the MAPKs with the selected downstream WRKY transcription factors. Therefore, we have generated useful resources in the form of entry clones, gateway compatible split-ubiquit in based Y2H vectors and the legume-specific MAPK cascade interaction data. In near future, the role of individual interactions in relaying the signals will be studied.
PD237 (IPPC0981) Non-targeted metabolomics approach to decipher biochemical mechanism under drought tolerance in rice (Oryza sativa. L) Pushpa Doddaraju1*, M.D. Prathibha1, Sumant Kumar Kambalimath1, A.B. Priyanka1, H.G. Jalendra Kumar2, H.V. Thulasiram2, M. Udayakumar1 and M.S. Sheshshayee1 1Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus, Bangalore-560065, Karnataka, India 2CSIR-Natioinal Chemical Laboratory, Pune-41008, India *Presenting author: [email protected] Rice cultivation account for over 50% of water utilized in agriculture and hence increasing water productivity is paramount significance. Though aerobic cultivation can save up to 60% of irrigation water, concomitant yield loss of upto 40% is often noticed is caused primarily due to an increased spikelet sterility. Cellular level tolerance (CLT) associated with production of compatible solutes and metabolites that reorganize proteins and cellular structures leading to maintenance of leaf turgor, cellular redox balance and homeostasis. Comprehensive profiling of solutes and metabolites in rice genotypes with varied level of drought tolerance and spikelet fertility would leaf to understanding the biochemical mechanisms at the cellular level and to identify genes responsible for high CLT under drought conditions. Fingerprint of biochemical perturbations could also be a reliable method to phenotype CLT under stress for breeding drought tolerant genotypes. In this study, differentially accumulated metabolites among drought tolerant and susceptible rice genotypes under well watered and water stress were measured using high resolution LC-MS. Metabolites that accumulated with significantly higher abundance in drought tolerant rice genotype compared to susceptible genotype upon stress induction were identified as drought tolerance related induced (DTRI) metabolites. Such DTRI metabolites mainly belonged to primary metabolites: sugars, amino acids and secondary metabolites such as phenylproponoids, flavonoids and alkaloids. Primary metabolites mainly involved in synthesis of stress responsive proteins and osmoprotection and secondary metabolites as antioxidants, reactive oxygen species (ROS) scavengers, coenzymes, UV and excess radiation screen and also as regulatory molecules.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 234 Session D : Plant Metabolism and Metabolomics
PD238 (IPPC1007) Health benefits and therapeutic properties of dietary rice bran Irfan A. Ghazi1*, Genevieve M. Forster2, Jan E. Leach3 and Elizabeth P. Ryan4 1Department of Plant Sciences, University of Hyderabad, Hyderabad-500046, India 2Department of Clinical Sciences, 3Department of Bioagricultural Sciences and Pest Management, 4Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, Colorado-80523, USA *Presenting author: [email protected] Rice is one of the most important cereal crops that feed more than half of the world population. Rice bran (RB) is a discarded by-product of rice milling, which was earlier used for animal feed, contain a variety of health promoting compounds. These compounds have chemotherapeutic, disease protective and several health beneficial properties. These properties include anti-oxidant, anti-cancer, anti-tumor and antidiabetic. RB from diverse varieties contains different bioactive compounds, such as polyphenol, tricin, ferulic acid, carotenoids, vitamin E, lutein and essential amino acids. RB Oil (RBO) is rich in gamma-oryzanol, phytosterol and sterolins. These compounds help in preventing oxidative damage of DNA, damage of body tissue, high blood pressure, hyperglycemia, hyperlipidemia and stimulate our immune system and protects against UV radiations. It has been proposed by several groups that RB may be used as functional food as it has cholesterol lowering properties, anticancer activity and has cardiovascular health benefits. RB derivatives are also used for different types of dermatological and cosmetic applications. Dietary RB intake also induced non- specific gut mucosal immune responses in mice. We have investigated how RB improves intestinal health by increasing innate resistance against enteric pathogens such as Salmonella that cause diarrhea in mouse model. RB from different colored and non-colored varieties can be used in breeding and transgenic to develop enteric disease protective traits in ongoing global rice crop improvement programs. Dietary RB is a promising food, which can be further investigated for its nutraceutical and therapeutic properties.
PD239 (IPPC1017) Towards understanding the biosynthesis and regulation of withanolides in Withania somnifera Dinesh A. Nagegowda1*, Anup Kumar Singh1, Varun Dwivedi1, Shaifali Pal2, Seema Meena1, Rajeev Kumar1 and Ajit Kumar Shasany2* 1Plant Molecular Biology and Biotechnology Lab, CSIR-CIMAP Research Centre, Bangalore-560065, India 2Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow-226015, India *Presenting author: [email protected] Ashwagandha (Withania somnifera L.) accumulates unique class of triterpenoid steroidal-lactones collectively termed as withanolides, which have diverse biological activities. To enable fast and efficient functional characterization of genes in this slow-growing and difficult-to-transform plant, a virus-induced gene silencing (VIGS) was established and its utilization was validated by silencing the gene encoding squalene synthase (SQS), which provides precursors for triterpenoids. VIGS of SQS resulted in significant reduction in squalene and withanolides, demonstrating its application in studying withanolides biosynthesis in W. somnifera leaves. Further, using VIGS approach a WRKY transcription factor (WsWRKY1) involved in regulation of withanolides accumulation in W. somnifera, was characterized. WsWRKY1 was induced by phytohormones, predominantly expressed in leaves, and localized to the nucleus. VIGS of WsWRKY1 in W. somnifera resulted in stunted plant growth and reduced withanolides, which was brought about by negative regulation of sterol pathway genes and reduced phytosterols. In contrast, transient over expression of WsWRKY1 in W. somnifera leaves up-regulated sterol pathway genes and enhanced the accumulation of withanolides. Moreover, ectopic expression of WsWRKY1 in tobacco led to up-regulation of sterol pathway genes with significant increase in stigmasterol and campesterol. Furthermore, while WsWRKY1-silencing led to down-regulation of defense related genes resulting in reduced tolerance to bacterial growth, fungal infection, and insect feeding, its over-expression in tobacco led to improved tolerance to biotic stresses. Our results demonstrated that WsWRKY1 could be a metabolic engineering tool to improve withanolides in W. somnifera plants or cultures and to enhance phytosterols and biotic stress tolerance in plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 235 Session D : Plant Metabolism and Metabolomics
PD240 (IPPC1037) Evaluation of flavouring compounds in local and processed potato products Pinky Raigond*, Brajesh Singh, Alka Joshi and Som Dutt Central Potato Research Institute, Shimla-171001, India *Presenting author: [email protected]
Market for processed potato products is rising day by day. Because of consumers preferences for better tasting foods, product flavor plays an important role in decision making by the consumers. In potato and potato products, glutamic acid, aspartic acid, guanosine 5’-monophosphate (GMP) and adenosine 5’-monophosphate (AMP) are the major umami compounds which contribute towards its flavour. Therefore, umami 5’ nucleotides (AMP+GMP) were estimated from local potato products (Samosa Fried, AlooTikki, Lachcha Fried, Lachcha Tokri and Aloo Seekh) as common fried products in the Indian markets and processed potato products (LAYS- Indian Magic Masala, Classic Salted, Swiss Grilled, Thai Sweet Chilli, American ASCO, Mccain- Potato Shortz, Masala Fries, Fries, Potato Chilli Bites, Smiles, AlooBhujia, Dehydrated- Potato Chips, Crax, Lachcha and Samosa Baked) being sold by retailers. The frozen products were fried before sampling. The analysis was done through HPLC using standardized protocol. Umami 5’ nucleotide content ranged from 2.63 (Alooseekh) to 8.26 (fried lachcha) μg g-1 fr. wt. in local potato products. Along with aloo seekh, fried samosa also contained low level of umami 5’ nucleotides, i.e., 4.42 μg g-1 fr. wt. In processed potato products, umami 5’ nucleotides ranged from 2.72 to 14.75 μg g-1 fr. wt. Umami 5’ nucleotides were the highest in aloo bhujia (14.75 μg g- 1 fr. wt.), followed by dehydrated aloo lachcha (11.14 μg g-1 fr. wt.) and dehydrated potato chips (10.13 μg g-1 fr. wt.). Processed potato products such as McCain Smiles (2.72 μg g-1 fr. wt.), McCain potato shortz (3.40 μg g-1 fr. wt.) and dehydrated potato crax (3.51 μg g-1 fr. wt.) contained low level of these compounds. The study suggests that the potato products prepared solely from potato contained higher levels of umami 5’ nucleotides compared to other products prepared by mixing potato with other cereals and vegetables.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 236 Session E : Biotic Stress Management and Interactions
PE241 (IPPC0064) Exploring a putative plant defensin from Rorippa indica and its integration into Brassica juncea for enhanced pest management Poulami Sarkar* and Samir Ranjan Sikdar Division of Plant Biology, Bose Institute, P-1/12, C.I.T. Scheme VII M, Kolkata- 700054, West Bengal, India *Presenting author: [email protected]
Mustard aphid - Lipaphis erysimi causes severe damage to the crop Brassica juncea not only by sucking the phloem sap but also by transmitting several disease causing viruses leading to an eventual yield loss from 35.4% to 91.3% annually. Previously, Rorippa indica - a wild crucifer was identified as aphid tolerant by the present group. To understand the tolerance mechanism of Rorippa indica - L. erysimi incompatible interaction, cDNA amplified fragment length polymorphism (cDNA AFLP) analysis was carried out in which a putative EST homologous to PDF1.2c - a defensin from Arabidopsis was identified. In the present study differential expression of this significantly upregulated EST has been studied using real time PCR analysis. The full length gene corresponding to the EST has been isolated and cloned from R. indica and named as Rorippa indica defensin (RiD). L. erysimi mortality in artificial diet based insect-bioassay with the in vitro expressed RiD indicated that the gene can be deployed for developing aphid tolerance in transgenic crops. The full length coding sequence of RiD was finally cloned into plant transformation vector pCAMBIA 1301. The pCAMBIA 1301 - RiD gene construct has been introduced into susceptible cultivar of B. juncea by using Agrobacterium-mediated plant transformation protocol with an aim to develop aphid tolerance. Molecular analyses of the putative B. juncea transformants are underway. B. juncea lines with successful integration and expression of RiD might provide enhanced aphid tolerance resulting in better pest management.
PE242 (IPPC0102) Potentials of Parthenium to control root knot nematode in cowpea Nisha Raghava*, Ravindra Pratap Raghava and Jayanti Srivastava FIST-DST Department of Botany, S.M.M. Town (PG) College, Ballia-277001, U.P., India *Presenting author: [email protected]
Cowpea (Vigna unguiculata (L.) Walp.) is a major pulse crops commonly used by Indians in a different ways as food and feed to cattle. It is most important crops as fodder and for green vegetable being rich in protein at large amount and many other nutrients. The roots of cowpea plants are severely galled by Root-Knot Nematode (RKN) at different stages of growth. Due to RKN formation, the growth of nodules is greatly affected and it causes damage to the plant growth. RKN are hidden enemies of crop and represent a unique challenge to agricultural research. Plant-parasitic nematode causes serious economic losses for many important agricultural crops and effective management are essential for sustaining modern agricultural productivity. Management techniques have relied on chemical control measures since their development during the middle of the last century. Regulatory restriction on the use of conventional nematicides now known to be human health and environmental hazards, necessitate the identification and evaluation of alternative non-chemical management strategies. The allelopathic potentials of Parthenium hysterophorus (L.) family Asteraceae have been utilized to control root knot nematodes in cowpea. The aqueous extracts of different parts of Parthenium plant were inhibitory to the number of eggs and active adult nematodes. The data collected on number of galls developed on the root surface showed remarkable reduction with different concentrations of Parthenium extracts, when sprayed at three stages of growth.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 237 Session E : Biotic Stress Management and Interactions
PE243 (IPPC0188) Induced defense responses in chilli pepper (Capsicum annum L.) against anthracnose pathogen Colletotrichum gloeosporoides Rukmini Mishra*, Ellojita Rout, Satyabrata Nanda and Raj Kumar Joshi Centre of Biotechnology, Siksha ‘O’ Anusandhan University, Bhubaneswar-751003, Odisha, India *Presenting author: [email protected]
Anthracnose, caused by Colletotrichum gloeosporoides is one of the most important diseases of Capsicum worldwide. In the present study, expression profiles of nineteen defense responsive genes were compared temporally during compatible and incompatible interaction with C. gloeosporoides in two capsicum lines. Quantitative real time PCR revealed that jasmonic acid (JA) and ethylene (ET) biosynthetic pathway related genes such as Plant defensin (PDF1.3), Lypoxygenase1 (Lox1), Allene oxide synthase (AOS) and ACC synthase 2 (ACS2) were rapidly induced and to a significantly higher level in the resistant line Bhoot Jolokia than in the susceptible cultivar Teja Jhal in response to anthracnose infection. While salicylic acid (SA) related gene phenylalanine ammonia-lyase (PAL) showed significant upregulation under incompatible interaction, isochorismate synthase (ICS2) showed similar expression for both genotypes. Further, three pathogenesis related proteins PR1, Chitinase (ChiIV) and PR5 were moderately responsive and showed similar expression patterns at all time points in both the genotypes. However, β-glucanase was highly expressive during compatible interaction. Additionally, transcription factors associated with disease resistance such as WRKY, MYB, ERF and bZIP were remarkably induced in the resistant genotypes as early as 3 dpi and remained significantly high at 9 dpi as compared to susceptible genotype. Our results indicated that capsicum adopts multiple strategies by activating several genes from various defense response pathways that play considerable role in resistance to anthracnose. The present study represents a valuable resource for future functional genomics experiments to unravel the molecular mechanisms of capsicum defense against C. gloeosporoides.
PE244 (IPPC0197) Parthenium leaf allelochemical preparation as an herbicide for noxious submerged aquatic weed hydrilla (Hydrilla verticilata L. f. Royle), a weed against weed Daya K. Pandey* Principal Scientist (Plant Physiology), ICAR-Directorate of Weed Research, Jabalpur-482004, India *Presenting author: [email protected]
Parthenium (Parthenium hysterophorus L.), one of the world’s worst weeds, continues to spread throughout the country affecting human and animal health, agriculture, environment and natural biodiversity with tremendous socio-economic implications. Phenolics, terpenoids and other constituents in it owe the species a range of biological activities including toxic and allelochemic interactions. Hydrilla (Hydrilla verticillata L. f. Royle) is a prominent submerged ubiquitous aquatic weed of waters with wide range of implications on the ecosystems and their services. Present study explores an herbicidal preparation from parthenium leaf for management of hydrilla. Parthenium leaf allelochemical herbicidal preparation (PLAHP) comprised of acetone water mixture extract of shade dried leaf powder. The PLAHP had about 12% parthenin and 4% phenolics. Majority of these were released instantly into the medium on suspending in water. Pre-weighed hydrilla plants were placed in suspensions of the PLAHP in 20% (v/v) nutrient medium. Biomass, toxicity, phenolics and terpenoids in the medium and herbicidal mechanism of action of the preparation were monitored. The experimental results were verified thrice by using bulk volume (40 litres) outdoors. The PLAHP was inhibitory at 400–500 ppm and herbicidal at 500-1000 ppm, killing the treated hydrilla plants in 5-10 days by involving fragmentation, loss of pigments and massive damage to cellular membranes, with a concurrent rapid dissipation of phenolics and parthenin in the medium. The herbicidal action of PLAHP on hydrilla opens up avenues for management of the weed as an eco-friendly approach by utilizing a weed like parthenium.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 238 Session E : Biotic Stress Management and Interactions
PE245 (IPPC0217) Concurrent overexpression of OsGS1.1 and OsGS2 for environmental stress resistance and herbicide tolerance in rice (Oryza sativa cv. japonica) Donald James*, Dhirendra Fartyal, Mrinalini Manna, Aakrati Agarwal, Baburam Verma, Bhabesh Borphukan, Mohan Achary and M.K. Reddy International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-1100167, India *Presenting author: [email protected]
Glutamine synthetase (GS; EC 6.3.1.2) is a key enzyme in the GS-GOGAT (Glutamine synthetase-Glutamate synthase) pathway controlling nitrogen metabolism in plants; it catalyzes the critical incorporation of inorganic ammonium into glutamine. Glutamine synthetase exists primarily as two isozymes with different subcellular localizations: GS1 in the cytosol and GS2 in chloroplasts/plastids. Several reports have implicated both cytosolic and chloroplastic GS isozymes in conferring resistance to various abiotic stresses in many crop species. Furthermore, glutamine synthetase is the target enzyme for commonly used non-selective herbicide Glufosinate/BASTA (active ingredient phosphinothricin; PPT). This study discusses the combined effect of constitutive overexpression of both OsGS1;1 and OsGS2 in rice with respect to tolerance to various abiotic stresses and resistance to BASTA. Two full-length cDNAs that encode the rice cytosolic glutamine synthetase1 protein (OsGS1;1) and the chloroplastic glutamine synthetase2 protein (OsGS2) were isolated from rice leaf tissue. The GS genes (OsGS1;1 and OsGS2) were cloned under rice actin constitutive promoters and pyramided invitro (stacked) into pMDC99 plant transformation vector via Gateway® technology. The OsGS1;1 and OsGS2 overexpressing rice (Oryza sativa cv japonica) transgenics were generated using Agrobacterium tumefaciens- EHA105 strain mediated transformation using hygromycin as a selectable marker. The putative transgenic plants were screened for transgene integration by PCR using hygromycin resistance gene (hptII) primers and further by Southern blotting. The transgenic plants exhibited limited tolerance to BASTA herbicide (Glufosinate/PPT) compared to wild-type plants as assayed by BASTA leaf paint test. The transgenic plants also showed considerable reduction in ammonium accumulation as determined by modified Berthelot assay.
PE246 (IPPC0244) Triple herbicide tolerant transgenic indica rice for combating weed problems Dhirendra Fartyal*, Aakrati Agarwal, Donald James, Mrinalini Manna, V. Baburam, Mohan Achary and M.K. Reddy International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India *Presenting author: [email protected]
Weeds are major biotic constraints in Indian as well as world agriculture systems. In addition to competition for nutrients and water, sometimes, they are alternate hosts for different kinds of pests and pathogens, which ultimately results in reduced crop production. So, it is necessary to eliminate weeds from the field to obtain higher food production. Since manual weed removal is expensive in terms of time, labour and money, it seems logical to use chemicals/herbicides which are very effective for weed destruction. But the use of a single herbicide continuously makes the weeds resistant to that particular herbicide in the long term. Hence, it is always beneficial to use different herbicides alternately. Keeping all these things in mind, we have developed triple herbicide tolerant indica rice (cv. Swarna). Three genes viz. EPSPS, ALS and BAR, along with a supporting gene igrA, were integrated to impart resistance to herbicides glyphosate, sulphonyl urea and glufosinate respectively. The igrA gene also helps to detoxify glyphosate along with EPSPS. All these genes were stacked in pMDC99 binary vector and transformed into EHA105 Agrobacterium strain. The rice calli were infected with these Agrobacterium cells and the transformed calli were selected with hygromycin selection. The selected calli were transferred to regeneration media and then rooting media to develop the transgenic plants. The putative transgenic plants were screened with PCR and southern blot and the confirmed transgenic plants were further subjected to respective herbicide assays.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 239 Session E : Biotic Stress Management and Interactions
PE247 (IPPC0251) Targeted mutagenesis of EPSPS gene Using CRISPR/Cas9 platform for the development of Glyphosat- resistant rice Bhabesh Borphukan* and V. Mohan Achary, Vijay Reddy, James Donald, Baburam Verma and M.K. Reddy Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Development of crops for withstanding marginal lands, resistant/tolerant for different biotic and abiotic stresses and bio- fortification for nutritional enhancement are the major area of concerned for food security in the coming years. Crop improvement through transgenic approaches has been arrested due to the continuous criticism on different ethical issues whereas cost of time and labour has to pay for conventional breeding. Recently CRISPR/Cas9 system has been emerged as a powerful tool for targeted genome editing. It provides opportunities for unprecedented manipulation of genetic materials either through gene knock out or knock in via non-homologous end joining (NHEJ) or homologous directed repair (HDR) pathway, respectively. CRISPR/Cas9 based HDR system provides a better platform for creating or changing phenotype in a desired way by altering specific DNA sequences, successful application of which will outclass both transgenic and conventional breeding approaches. Here we have developed Cas9 expressing rice platform under the control of Zm ubiquitin promoter. This platform was further used for creating T169I and P173S mutation through homologous recombination in the native OsEPSPS to make glyphosate tolerant indica rice variety Satabdi. Successful demonstration of Cas9 based HDR mediated genome editing promise efficient and precise engineering of crop plants in near future through editing native gene or regulatory elements which also assure significant improvement of crop genetic material in a cisgenic approaches which could be accepted more by the consumers in near future.
PE248 (IPPC0279) Tolerance assessment of transgenic Brassica overexpresing MPK3 gene against Alternaria blight Mohd Hassan1, Mohd Tasleem1, Dinesh Pandey1, Anil Kumar1, Vandna A. Kumar2 and Gohar Taj1* 1Depatment of Molecular Biology and Genetic Engineering, 2Department of Biochemistry, G.B. Pant University of Agriculture and Technology Pantnagar-263145, India *Presenting author: [email protected]
The potential yield of Brassica in India is very high but there is a wide gap between the potential and realized yields, which is largely because of the biotic and abiotic stresses. Alternaria blight is the most important diseases of rape seed mustard that leads to major yield losses as well as deterioration in quality of oilseeds. Therefore, in the present study, transgenic Brassica (BjV2) plant over-expressing MPK3 gene was studied to assess tolerance against Alternaria blight. Lower disease index and higher antioxidant content found in the transgenic plant (BjV2) as compared to wild Brassica suggests its tolerant capability to Alternaria blight disease. The similar expression pattern of MPKK9 and MPK3 suggests that MPKK9 might be an upstream activator of MPK3 and work in a direct cascade for the production of defense related proteins. MPKK5 was found to support the colonization of the pathogen, as the wild plant showed more expression than the tolerant transgenic (BjV2) plant. The expression of WRKY transcription factors was also studied, and revealed that WRKY33 acts downstream of MPK3 in activating defense related genes. With lower disease index, higher ROS scavenging enzymes content the transgenic Brassica (BjV2) was found to be tolerant to the pathogen Alternaria brassicae.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 240 Session E : Biotic Stress Management and Interactions
PE249 (IPPC0306) Identification of Bacillus sp. from algae culture system and its effect on growth of Desmodesmus ultrasquamatus Yash Gupte*, Ganesh Iyer, Suhas Pednekar, Vinod Nagle, Sailee Lavekar and Pooja Devadiga Ramanarain Ruia College, Matunga, Mumbai-400019, Maharashtra, India Presenting author: [email protected]
Bacillus sp. was isolated from algal culture systems. Presence of bacteria in algal culture system may be a mutually benefitting relationship. Bacillus being a facultative anaerobe can grow well in the oxygen rich water of algae and in turn help algae to continue photosynthesis. It often forms a film over the surface of water if the media is not mixed thoroughly. The gram character was studied to identify the bacterium morphologically as Bacillus coagulans. PCR amplification of 16s rDNA gene was carried out and the sequence was analyzed using BLAST for molecular identification of the bacterium. Bacillus sp. was found to be resistant to Ampicillin, Kanamycin and Neomycin which may be due to the endospore forming capacity and change of gram character when passing into stationary phase. The associative growth of algae and bacteria was found to enhance algal growth by 7-9%, with no difference in the biochemical characters of algae Desmodesmus ultrasquamatus. Gas Chromatography Mass Spectroscopy (GC-MS) revealed very little change in the fatty acid profile of the Desmodesmus ultrasquamatus.
PE250 (IPPC0317) Host-induced gene silencing in tomato and chilli against anthracnose disease caused by Colletotrichum gloeosporioides Binod Kumar Mahto* and Swatismita Dhar The Energy and Resources and Institute (TERI) University, Vasant Kunj, New Delhi-110070, India *Presenting author: [email protected]
Anthracnose disease is caused by ascomycetes fungus, Colletotrichum spp. which is responsible for heavy yield loss for tomato and chilli production around the worldwide. We are reporting the expression of RNA interference (RNAi) sequences derived from an essential Colletotrichum gloeosporioides virulence gene, Conidial Morphology 1 (CgCOM1) involved in fungal conidial and appresorium development, as a method to enhance resistance of tomato (var. Pusa Rohini and Pusa Early Dwarf) and chilli (var. Pusa Jwala and Pusa Sadabahar) plants against Colletotrichum sp. We have developed transgenic lines of tomato (Solanum lycopersicum) and chilli (Capsicum annuum) using RNAi construct containing pathogen gene by Agrobacterium-mediated stable transformation. We have already standardized the Agrobacterium-mediated stable transformation protocol in tomato and chilli with pCAMBIA2301 vector. We show that HIGS (Host-Induced Gene Silencing) can be used to silence expression of pathogen genes involved in general function which can confer resistance against fungal pathogens.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 241 Session E : Biotic Stress Management and Interactions
PE251 (IPPC0333) Study on biotic stress interaction on application of a potential herbicidal fungus, Gibbago trianthemae Pinki Bhandari*, Meenakshi Agarwal and S.K. Raza Bioscience Division, Institute of Pesticide Formulation Technology, Gurgaon-122016, India *Presenting author: [email protected]
Gibbago trianthemae is a fungus, with potential bioherbicidal activity when applied at high densities onto weed, viz., Trianthema, and showed 70-75% weed control. The present study revealed that Gebbago trianthemae working as phyto- pathogen fungi via selective host specific phytotoxine significantly affect only the plant species Trianthema portulacastrum of family Aizoaceae. Under biotic stress there is production of singlet oxygen and other photodynamic compounds, such as hypericin. Plant pathogens produce a variety of phytotoxins that interfere with plant metabolism, ranging from subtle effects on gene expression to plant mortality. Solutions of Gibbago trianthemae, diluted to different concentrations were applied in the field with three replications. Soil samples were collected from 7 consecutive depths in (0-15 cm, 15-30 cm, 30-45 cm, 45-60, 60-75 cm, 75-90 cm, 90-120 cm) of soil profile using 5 cm diameter soil core. The soil samples were enumerated for different group of micro-organisms using different growth media. Collected soil samples diluted and spread on medium plates, and incubated for 24-48 h, and the growth of micro organism was calculated. It was found that the Gibbago trianthemae has non-significant effect on soil bacteria and actinomycetes growth.
PE252 (IPPC0344) Effect of mycoherbicide fungus Gibbago trianthemae for the control of carpet weed & its impact on certain rabi crops Meenakshi Agarwal*, P. Bhandari and S.K. Raza Bioscience Division, Institute of Pesticide Formulation Technology, Gurgaon-122016, India *Presenting author: [email protected]
The recent years there has been a increasing interest in mycoherbicide fungi caused by factors such as increasing insecticide resistance and environmental concerns over pesticide use. The changing environmental conditions and increasing demand & consumption of chemical insecticide has created biotic stress in overall agro ecosystem of major economics crop viz. In present study Gibbago trianthemae simmons a phaeodictyoconidal hyphomycetous fungus reported on carpet weed, causes leaf spot disease resulting in premature defolation. In present study, a highly virulent fungal pathogen strain. Gibbago trianthemae was screened out on the basis of its pathogenic potential to develop sporulating lesion on horse purslane. Seedling/seeds of major rabi crops were planted in the plots. It is liquid suspension formulation with 1x106 spores ml-1 was found to be most pathogenic on horse purslane. These crops were sprayed with the Gibbago trianthemae suspensions. Observation on disease incidence and plant vigour were recorded after 5,10,15 and 30 days after application. There is no disease incidence of Gibbago trianthemae on rabi crops. In host range studied Gibbago was found pathogenic only to horse purslane and there was no phytotoxicity and any other biochemical change observed on the main crop viz. cabbage, cauliflower, tomato, chilli, fenugreek, maize, mustard, coriander. It was found that the Gibbago trianthemae has no significant effect on soil bacterial and actinomycetes growth i.e. (4.44 x 105 CFU/ml) and (3.61x 105 CFU/ml) respectively. Whereas a significant effect on soil fungi at 1.87 x 105 CFU/ml it was also negligible but significant effect.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 242 Session E : Biotic Stress Management and Interactions
PE253 (IPPC0431) Understanding the molecular intricacies of Rice-Rhizoctonia solani interactions Srayan Ghosh*, Poonam Kanwar and Gopaljee Jha National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Rhizoctonia solani is a necrotrophic soil borne pathogen causing sheath blight disease in rice and is responsible for huge crop loss every year worldwide. In spite of global efforts, till now there have been no reports of even a single R gene identified against this pathogen. Hence understanding the molecular intricacies of Rice-Rhizoctonia is required to device better strategies to control this important disease. While mining the available genome sequences of rice and lettuce isolate of R. solani belonging to AG1-IA and AG1-IB anastomosis groups, respectively we have identified several genes that are exclusively present in AG1-IA. Many of them encode important biological, molecular functions and exhibit dynamic expression during in-planta growth of the pathogen in rice. Based upon sequence similarity with genes that are required for plant and human/zoonotic diseases (PHIbase database), several putative virulence/pathogenicity determinants amongst AG1-IA specific genes were identified. Further the differentially expressed pathogen and host genes during Rice-Rhizoctonia interactions have been catalogued while analysing the RNAseq data. We endeavour to understand how R. solani alters the host’s physiological environment to create a suitable niche to establish itself and cause necrosis.
PE254 (IPPC0442) The wheat resistance gene Lr34 results in the constitutive induction of multiple defense pathways in transgenic barley Harsh Chauhan1*, Rainer Boni2, Liselotte Selter2, Thomas Wicker2, Simon Krattinger2 and Beat Keller2 1Indian Institute of Technology (IIT) Roorkee-247667, Uttarakhand, India 2University of Zurich, Switzerland *Presenting author: [email protected]
The wheat gene Lr34 (Yr18/Pm38/Sr57/Ltn1) encodes a putative ABCG-type of transporter and is a unique source of disease resistance providing durable and partial resistance against multiple fungal pathogens and has been used in modern wheat breeding for more than 100 years. Lr34 has been found to be functional as a transgene in barley but its mode of action and biochemical function remained largely unknown both in wheat and barley. Here, we studied gene expression in uninfected barley lines transgenic for the Lr34 gene. Multiple defense pathways contributing to basal as well as inducible disease responses such as lignin synthesis, phytoalexin biosynthesis and pathogenesis-related protein genes were constitutively active both in seedling and mature leaves. In addition, the hormones jasmonic acid and salicylic acid were induced to high levels in the presence of Lr34. We found that in barley the deletion of a single phenylalanine residue was sufficient to induce the characteristic Lr34-based responses. Expression of Lr34res alone resulted in a severe reduction of plant vigor in transgenic barley. These negative effects of Lr34res on barley growth were attenuated in the presence of the susceptible Lr34sus allele, whereas resistance function was not compromised. These results demonstrate a strong re-programming of plant metabolism by Lr34 even in the absence of the pathogen. Interestingly, the negative effects on growth resulting from defense gene induction can be attenuated genetically, suggesting that a transgenic combination of Lr34res and Lr34sus in heterologous grass species might result in agronomically useful resistance.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 243 Session E : Biotic Stress Management and Interactions
PE255 (IPPC0479) Tomato miR159-SlMyb33 interface modulates R gene activity to control virus infection Pranav Pankaj Sahu, Manoj Prasad and Namisha Sharma* National Institute of Plant Genome Research, Aruna Asif Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Tomato Leaf Curl New Delhi Virus (ToLCNDV), a bipartite geminivirus, causes Tomato leaf curl disease and adversely affects the horticultural economy. Natural defense system against geminiviruses includes the phenomenon of RNA interference. To study pivotal role of miRNAs in plant defense against ToLCNDV, two small RNA libraries were constructed in mock treated and ToLCNDV infected cultivar H-88-78-1. Some of the identified known and novel miRNAs were analyzed by northern blot for their differential expression in two contrasting cultivars (tolerant cv. H-88-78-1 and susceptible cv. Punjab Chhuhara) of tomato, among which 13 miRNAs showed differential expression in both the cultivars. After viral infection down-regulated expression was observed for sly-miR159 in tolerant cultivar and showed inverse relationship with its target SlMyb33, which was also validated by RLM RACE. Silencing of SlMyb33 in cv. H-88-78-1 was performed to determine the plausible role of SlMyb33 during plant-virus interaction, which revealed that ToLCNDV-infectivity and viral titer were significantly increased in silenced line in comparison to cv. H-88-78-1 innate state. Promoter binding characteristic of known virus resistant genes was further explored for Myb33 binding sites. In silico study of promoters of eight resistant genes revealed that three of them (N gene, Sw-5, and Tm2) have Myb33 binding sites. Further, interaction between Myb33 and promoter of differentially regulated R gene was validated through EMSA. Till now, miR159 have been shown to be associated with the symptom development upon virus infection, we hypothesize that upon viral infection reduced expression of sly-miR159 leads to higher accumulation of its target SlMyb33, which in turn activate the transcription of the R gene and provide tolerance against ToLCNDV.
PE256 (IPPC0504) Functional involvement of a group C mitogen activated protein kinase, OsMPK7 in mediating resistance against Xanthomonas oryzae in rice Siddhi Jalmi* and Alok Sinha National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Mitogen-activated protein kinases (MAPKs) are highly conserved signaling cascades in eukaryotes, involved in transducing stress related signals from upstream receptor to downstream target by a phosphorelay mechanism. In this study the role of OsMPK7 is assessed during Xanthomonas oryzae infection, a causal agent of leaf blight disease in rice. X. oryzae treatment resulted in induction of transcripts of OsMPK7 and OsMKK3, as well as activation of OsMPK7. Further, OsMPK7 was found to be target of OsMKK3 and the two proteins interacted in-planta. Overexpression of OsMPK7 and OsMKK3, individually and in combinations resulted in inhibition of disease symptoms caused by X. oryzae, both in leaf and root. Furthermore, the mechanism for disease resistance was exemplified by studying relative expression of defence related genes in rice overexpressing OsMKK3-OsMPK7 module. This revealed that overexpression of OsMKK3-OsMPK7 upregulated pathogenesis-related genes and genes involved in cell wall metabolism. These leaves also showed restricted movement of the pathogen from the point of infection to other uninfected part. To explore downstream target of OsMPK7 involved in mediating X. oryzae related cues, protein-protein interaction studies were performed. OsWRKY30 was identified as a putative downstream target of OsMPK7. Taken together, this work suggests a positive involvement of OsMKK3- OsMPK7 module in imparting disease resistance against X. oryzae infection in rice. Further, the role of OsWRKY30 in OsMKK3-OsMPK7 mediated signaling in response to X. oryzae pathogen will be evaluated.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 244 Session E : Biotic Stress Management and Interactions
PE257 (IPPC0523) A DNA satellite encoded protein interferes with structure and function of chloroplast leading to photosynthetic inhibition Prabu Gnanasekaran*, Dhriti Bhattacharyya, Reddy Kishore Kumar, Nirbhay Kumar Kushwaha, Veerendra Kumar Sharma and Supriya Chakraborty School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Gemini viruses are known for their potency to interfere with plant’s physiology. Recently betasatellite, a group of DNA satellite has emerged as a major threat to crop production worldwide and acts as a key regulator for symptom induction. To study host-betasatellite interactions and symptom development, we used Nicotiana benthamiana - Radish leaf curl betasatellite (RaLCB) as model system. RaLCB initiates leaf curling, vein clearing and severe veinal chlorosis in infected plants; however, in the absence of βC1 protein, RaLCB failed to induce vein clearing symptoms on host plant. In the course of infection, βC1 protein localizes into chloroplasts of the host cells and damages chloroplast ultrastructure. In addition, the betasatellite downregulates expression of genes involved in chlorophyll biosynthesis, chloroplast development and plastid translocation. Interestingly, the expression of key host genes involved in chlorophyll degradation remains unaffected indicating selective suppression of host metabolism. Betasatellite infection drastically reduces the numbers of active reaction centres and plastoquinol pool size in leaves exhibiting vein clearing symptom. Betasatellite mediated impediments at different levels of chloroplast functionality affects photosynthetic efficiency of N. benthamiana. This study provides the first evidence of chloroplast localization of a DNA virus encoded protein which in turn affects photosynthesis.
PE258 (IPPC0529) Identification and characterization of a novel mycophagous bacterium against Rhizoctonia solani, causal agent of sheath blight disease of rice Isha Tyagi*, Rajeev Kumar and Gopaljee Jha National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Bacterial Mycophagy is a set of phenotypic behaviours that enable bacteria to obtain nutrients from living fungi allowing conversion of fungal biomass into bacterial biomass. We have isolated a novel yellow-pigmented bacterium from surface sterilized healthy rice seedlings demonstrating mycophagous behaviour against Rhizoctonia solani (the causal of sheath blight disease of rice), and broad spectrum antifungal activity against major phytopathogens such as Magnaporthe oryzae (the rice blast pathogen), Venturia inaequalis (the apple scab pathogen), Fusarium oxysporum (vascular wilt pathogen), and several other fungal pathogens. 16S rRNA gene sequencing, deep phylogeny, and segregation rooting analyses identified the bacterium to be Burkholderia gladioli, and henceforth, the strain was named Burkholderia gladioli NGJ1. The B. gladioli NGJ1’s mycophagous behaviour against R. solani is shown by bacterial growth over fungi, weight loss and cell death assays. The R. solani on co-inoculation with B. gladioli NGJ1 could not cause disease on rice tillers and tomato leaves thus suggesting that mycophagous phenomenon could be used as a strong biocontrol agent for sheath blight disease of rice by R. solani and several other economic important plant diseases. The draft genome sequence of B. gladioli NGJ1 has been unravelled and the bacterium is enriched in genes encoding many fungal cell wall degrading enzymes and several novel antibiotics.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 245 Session E : Biotic Stress Management and Interactions
PE259 (IPPC0532) Interaction of Allium sativum leaf agglutinin (ASAL) with GroEL related receptor interferes with whitefly mediated tomato leaf curl New Delhi virus (ToLCNDV) transmission Ayan Das1*, Amit Roy2, Arunava Mandal1, Pallob Kundu1 and Sampa Das1 1Division of Plant Biology, Bose Institute, P 1/12 C. I. T Scheme VII M, Kolkata- 700054, West Bengal, India 2Department of Plant Protection Biology, SLU, PO 102, SE-230 53 Alnarp, Sweden *Presenting author: [email protected]
Allium sativum leaf agglutinin (ASAL), a mannose binding lectin has been established as a promising insecticidal component. Its efficacy was checked on an array of hemipteran insects both in artificial diet based and in planta bioassay on ASAL transgenic plants. A unique receptor of ASAL was previously identified by the present group in brush border membrane vesicle (BBMV) of mustard aphid (Lipaphis erysimi) which was found to be GroEL related chaperonin protein- symbionin. GroEL has been reported to play role in persistent, circulative transmission of plant viruses. Both in silico and in planta experiments proved the plausible role of ASAL in interference with aphid mediated viral transmission. In the present work similar GroEL receptor of ASAL was identified through LC MS/ MS analysis in the BBMV of whitefly, the widely known vector for several economically important plant viruses including tomato leaf curl New Delhi virus (ToLCNDV). Ligand blot analyses demonstrates- when whiteflies are pre-fed with ASAL it completely blocks the binding of the ToLCNDV coat protein to GroEL. Co-localisation of ToLCNDV coat protein and GroEL was detected in whitefly through confocal microscopic analyses but ToLCNDV coat protein did not co-localise when insects were prior fed with ASAL. Immunoprecipitation followed by western analyses also showed that when whiteflies are pre-fed on ASAL supplemented diet it interferes with the binding of ToLCNDV coat protein to GroEL. These results confirm that ASAL’s interaction with GroEL receptor interferes with binding of ToLCNDV coat protein and subsequent whitefly mediated viral transmission.
PE260 (IPPC0535) Drought stress endures Arabidopsis thaliana plants to subsequent Pseudomonas syringae infection due to priming of basal defence responses Aarti Gupta*, Sandeep Dixit and Muthappa Senthil-Kumar National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India *Presenting author: [email protected]
The global climate change is predicted to cause frequent incidences of concurrent abiotic and biotic stresses. Recent studies indicate that plant responses are adaptively tailored for combating the concurrent stresses, which are otherwise not elicited under individual stress conditions. Drought potentially influences the occurrence and intensity of a particular disease by modulating the plant-pathogen interaction. Thus understanding the plant response towards concurrent drought and pathogen stressors is important. In order to facilitate such studies an accurate protocol for concurrent imposition of drought and pathogen is warranted. The prolonged and gradual nature of drought stress imposition protocol poses a hindrance in overlaying with pathogen infection in laboratory conditions to achieve combined stress imposition. Till date an accurate and convenient methodology is lacking, even in model plants like Arabidopsis thaliana. Here we comprehensively studied impact of drought stress on A. thaliana-Pseudomonas syringae pv. tomato DC3000 interaction. In this study we established a systematic protocol to study the impact of combined drought and bacterial pathogen stress. We also attempted to dissect the contribution of individual stress effect in this stress combination and showed that plant responses under combined stress are skewed towards the most prominent stressor, namely drought. We also observed that physiological responses under combined stress are tailored when compared to the plants exposed to individual stresses. Further, our results showed that drought stressed A. thaliana plants better defend oncoming pathogen infection. Both the developed protocol and results from the stress interaction are presented.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 246 Session E : Biotic Stress Management and Interactions
PE261 (IPPC0537) Identification of differentially expressed genes under combined drought and Ascochyta rabie infection in chickpea plants using meta-analysis and their functional validation Prachi Pandey*, Anjugam Paramanantham, Praveen Verma and Muthappa Senthil-Kumar National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India *Presenting author: [email protected]
Under field conditions, plants are simultaneously exposed to a number of abiotic and biotic stress conditions. Plants’ response to combined stress consists of a combination of shared and unique responses. Whereas shared responses are the responses common to the individual and combined stress conditions, unique responses are characteristically seen under combined stress conditions. The transcriptomic information available from the large number of individual abiotic and biotic stress related microarray datasets can provide valuable insights into the shared responses under the combined stress conditions. The present study aimed at understanding the shared molecular response of Cicer arietinum under combined drought and Ascochyta rabiei infection by meta-analysis of microarray data obtained from individual drought and A. rabiei infection experiments. The study identified a list of differentially expressed genes in chickpea plants which were common under drought and a fungal pathogen infection. Among all the commonly regulated genes, those encoding for pathogenesis related factor (PR4), carboxytransferase, linoleate 9 S-lipoxygenase like protein were some of the commonly upregulated genes. On the other hand, genes encoding for phosphate induced protein, proline oxidase and photosystem reaction centre proteins were found to be commonly downregulated under drought stress and A. rabiei infection. Data for the functional validation of few of these genes will be presented. Overall this study presents validation of the meta-analysis approach and conclude that re-analysis of the transcriptomic data of plants under individual stresses can provide insights into the shared mechanisms behind tolerance to combined drought and A. rabie infection in Cicer arietinum.
PE262 (IPPC0543) Identification and comparative analysis of RNAi suppression activity of different viral proteins Abhishek Anand*, Sudhanshu Sekhar, Veena Sinha and Neeti Sanan-Mishra International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
RNA silencing suppressors have emerged as a relatively new challenge to plant defense system against viruses. The viruses neutralize the plants defense strategy of RNA silencing by using the suppressor proteins. These proteins are already present inside the virus, performing their vital functions and have possibly developed the suppression activity during later stages of evolution.This explains the fact that they do not share any evolutionary homology or pattern. They are diverse not only in sequence but also in the ways they suppress the silencing machinery. So it has become difficult to predict any pattern based on their nucleotide or protein sequence or structure to identify and classify the suppressor proteins. Here we present a simple, quick and reliable strategy to identify a protein with suppressor function. This assay was used to identify RNAi suppressors from diverse viruses. It is now emerging that a single virus may contain multiple suppressors, each with a different mechanism of action. This assay can also quantitate the strength of suppression. The knowledge gained will complement the studies on their mechanism of suppressor action to understand the strategy adopted by the viruses to evade the host RNAi defense.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 247 Session E : Biotic Stress Management and Interactions
PE263 (IPPC0546) Molecular and physiological studies reveal complexities of rice Rhizoctonia solani interactions Poonam Kanwar*, Srayan Ghosh and Gopaljee Jha National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Rice, one of the world’s most important crops faces severe yield loss, due to sheath blight disease caused by a necrotrophic fungal pathogen Rhizoctonia solani. No source of complete disease resistance is known, till date. We adopted a multi- pronged approach to study the intricacies of plant pathogen interaction. The interaction transcriptome was explored to understand the dynamic interplay between host and pathogen genes and processes during infections. The alteration in photosynthesis, carbohydrate metabolism and induction of cell death were the most prominent host responses. This was further validated by physiological and biochemical analysis. Collectively, both transcriptome and functional assays revealed that R. solani causes extensive metabolic re-programming inside the susceptible host. In conclusion, the present study provides better understanding of this important patho-system and lays foundation for facilitating research towards developing strategies to combat the disease.
PE264 (IPPC0550) Artificial microRNA-mediated silencing of ecdysone receptor gene affects growth and development of Helicoverpa armigera Sneha Yogindran* and M.V. Rajam Department of Genetics, University of Delhi, South Campus, New Delhi-110021, India *Presenting author: [email protected]
Food production is seriously affected by various biotic stresses such as insect pests, nematode parasites, pathogens and weeds. Hence, it is necessary to increase the crop production to feed the ever growing population. Among the different biotic stresses, insect pests affect the crop productivity to a great extent. Helicoverpa armigera, popularly known as cotton bollworm is one of the serious agricultural pests which affect more than 360 plant species including 50 crop plants. This pest is a great threat to the farmers due to its host range, high mobility and fecundity, and the ability to adapt and develop resistance against all common groups of insecticides. Bt technology paved the way for this problem to a great extent, but reports came up with insects gaining resistance against Bt toxins. Therefore, there is an urgent need for an effective and long lasting pest control strategies. The present study focuses on the use of artificial microRNA (amiRNA) to target ecdysone receptor (EcR) gene of H. armigera. The insect steroid hormone ecdysone and its receptor play important roles during development and metamorphosis and regulate adult physiology and life span, making it a potential target for pest control. Insect feeding bioassays with amiRNA-HaEcR caused significant reduction in target gene transcript level and affected the growth and development of the insect when compared to control. These results suggest that the use of amiRNA technology may serve as a potential alternative to the current pest management practices.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 248 Session E : Biotic Stress Management and Interactions
PE265 (IPPC0557) Identification of ZFD1 interacting factors (ZIFs) which influence immune responses in Arabidopsis thaliana Anupriya Singh*, Nidhi Singh and Ashis Kumar Nandi School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Pathogens put a significant challenge on plant fitness and survival. To combat this challenge plants have evolved well developed immune system. Plant hormones such as salicylic acid (SA), ethylene (ET) and jasmonic acid (JA) play incredible role in plant defense. Infection with biotrophic pathogen results in accumulation of SA. SA binds to and activates non expressor of PR1 (NPR1). NPR1, the master regulator of SA signaling, in turn induces expression of large number of pathogenesis related (PR) gene. Many PR gene products are antimicrobial, which help in warding off the pathogens. Zinc Finger protein involved in disease Defense (ZFD1) another positive regulator of disease defense, recently identified in our lab (unpublished data). ZFD1 mostly influences SA pathway in Arabidopsis. By yeast two hybrid (Y2H) screening several ZFD1 interacting factors (ZIFs) were identified. Some of these ZIFs contain nuclear localization signal. Here we report two ZIFs, such as ZIF1 and ZIF2 which influence immune responses. Through bimolecular fluorescence complementation (BiFC) assay we showed that interaction of ZFD1 with ZIF1 takes place in the nucleus. Disease resistance studies using Pseudomonas syringae (Psm), we report here that ZIF1 is a negative regulator and ZIF2 is a positive regulator of immune response. In addition, we also report a third interacting partner ZIF3 which interacts with ZFD1 in ER. Role of ZIF3 in pathogenesis is under investigation.
PE266 (IPPC0586) Role of Arabidopsis Polycomb group repressor -2 complex in pathogenesis Priya Gupta*, Shweta Roy and Ashis K. Nandi School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Plants possess highly sophisticated immune response to combat pathogens. They have broader and basal defense response called pattern triggered immunity (PTI) as well as much more efficient and specific immunity called effector triggered immunity (ETI). Sometimes plants recruit housekeeping genes or development regulators for participating in defense. Through transcriptome analysis of WT Arabidopsis plants after HR induction, our lab identified a gene belonging to PRC2 complex, which supports bacterial growth. We named here as POLYCOMB REPRESSOR FOR DISEASE 1 (PRD1). In PRD1 overexpression plants, disease resistance gene Resistance to Pseudomonas syringae 2 (RPS2) expression is reduced. The amiRNA lines of PRD1 are resistant against Pseudomonas syringae pv tomato DC3000 carrying AvrRpt2 gene. We show here, by transient expression in Nicotiana benthamiana that PRD1 suppresses expression of RPS2:GUS. We have also analyzed the promoter of PRD1. Our results identified the region of PRD1 promoter that is pathogen inducible. Yeast-2-hybrid screening using PRD1 as bait identified a large number of putative interactors of PRD1. VOLTAGE DEPENDENT ANION CHANNEL 2 (VDAC2) and ABI FIVE BINDING PROTEIN 3 (AFP3) are two such putative interactors. Through BiFC we have reconfirmed and identified their sub-cellular location of interaction. Mechanism of PRD1 mediated disease susceptibility is under investigation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 249 Session E : Biotic Stress Management and Interactions
PE267 (IPPC0587) Overexpression of a rice A20/AN1 zinc-finger protein modulates defence response against pathogen infection in tobacco Shweta Jha1*, Meenakshi Sharma2, Jitender Giri2 and Akhilesh K. Tyagi2 1Biotechnology Unit, Centre of Advance Studies in Botany, Jai Narain Vyas University, Jodhpur-342001, Rajasthan, India 2National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India *Presenting author: [email protected]
Eukaryotic A20/AN1 zinc-finger domain containing proteins play an important role in the regulation of immune responses. In plants, Stress Associated Proteins (SAPs) are identified as novel A20/AN1 zinc-finger proteins, which are shown to be regulated by multiple abiotic and biotic stresses and emerging as potential candidates to confer abiotic stress tolerance in transgenic plants. A total of 18 genes were identified from rice genome encoding SAPs. Overexpression of SAPs from rice and other plants has been shown to confer abiotic stress tolerance in transgenic plants. In the present study, the functional role of OsSAP1 in plant defence response has been explored through its constitutive overexpression in transgenic tobacco plants. OsSAP1 overexpressing tobacco plants showed enhanced basal resistance against virulent bacterial pathogen, Pseudomonas syringae pv. tabaci, together with the upregulation of known defence-related biotic-stress responsive genes. Enhanced disease resistance is coupled with increased ROS production in OsSAP1 overexpressing plants. Although its mechanism of action is poorly understood, emerging evidences suggest that OsSAP1 may act as ubiquitin ligase, redox sensor and regulator of gene expression during stress. Thus OsSAP1 may be considered as an ideal candidate for protecting crop plants against multiple stresses and yield loss.
PE268 (IPPC0599) Responding rapidly to insect herbivores: calcium signalling in plant defense against herbivores Jyothilakshmi Vadassery1*, Sandra Scholz2, Victoria Kiep3, Edgar Pieter3 and Axel Mithöfer2 1National Institute of Plant Genome Research (NIPGR), Aruna Asaf Ali Marg, New Delhi-110067, India 2Department Bioorganic Chemistry, Max Planck Institute for Chemical Ecology, Hans-Knöll-Stra&szl 3Martin Luther University of Halle-Wittenberg, 06099 Halle (Saale), Germany *Presenting author: [email protected]
In the interaction between Arabidopsis and the generalist herbivorous insect, Spodoptera littoralis, little is known about early events in defense signaling and link to downstream phytohormone pathways. We demonstrate that cellular calcium (Ca2+) elevation is involved at multiple stages of interaction between S. littoralis and Arabidopsis in both local and systemic leaf. Upon insect herbivory S. littoralis oral secretions (OS), and bioactive jasmonic acid-isoleucine conjugate, induce cytosolic Ca2+ elevation in Arabidopsis. S. littoralis feeding also induces systemic [Ca2+]cyt elevations in Arabidopsis which are predominantly in leaves with direct vascular connections to the treated leaf and appeared with a delay of 1 to 2 minutes. Simulated herbivory by wounding always induced a local [Ca2+]cyt response, but a systemic one only when the midrib was wounded. This systemic [Ca2+]cyt response was suppressed by the presence of insect-derived oral secretions as well as in a mutant of the vacuolar cation channel TPC1. Using microarray, gene expression induced by Spodoptera oral secretion in Arabidopsis leaf was identified. Calmodulin like proteins (CML), CML42 and CML37, are highly up- regulated both by oral secretions and insect herbivory in local leaves. Functional analysis of CML42 and CML37 loss-of- function mutants revealed that they act in antagonistic manner during herbivory defense and modulate jasmonate pathway. The current study highlights that Ca2+ signaling in insect herbivory is an intricate and complex network with multiple components involving positive and negative regulators in response to local and systemic Ca2+ elevations and has a direct impact on jasmonate signaling.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 250 Session E : Biotic Stress Management and Interactions
PE269 (IPPC0642) Analysis of the genes involved in terpeneoid pathway during infection in Aquilaria malaccensis Md Rofiqul Islam* and Sofia Banu Bioengineering and Technology, Gauhati University, Guwahati-781014, Assam, India *Presenting author:[email protected]
Agarwood is a non-timber dark resinous wood that is found in the stems, branches, and roots of Aquilaria and Gyrinops trees after they have been wounded or exposed to biotic damage. Resin production is considered as one of the plant defence mechanisms resulting in accumulation of agarwood oil. Agarwood is widely used in traditional medicines, as a digestive, sedative, and anti-emetic and also as an incense and perfume, and this huge demand for the agarwood products has led all Aquilaria spp. being endangered and listed in the Appendix II of the CITES. To promote the production efficiency and protect the resource of Aquilaria malaccensis, it is critical to understand the regulation mechanisms of stress-induced agarwood formation. As the major components of agarwood are sesquiterpenes and phenylethylchromones, the present study aims to understand the molecular basis of wound-induced sesquiterpenes biosynthesis and agarwood formation. Since, the biosynthesis of sesquiterpenes occurs via the MVA and DXP pathways, involving the following enzymes in their synthesis, viz., 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR), DXP synthase (DXPS), farnesyl diphosphate synthase (FPS), and sesquiterpenes synthases (SS). The study presents the results of expression profiles of these four key enzymes as well as other terpenoid genes in different tissues (wood and leaf) of infected and non- infected A. malaccensis plants, analyzed using semi-quantitative PCR and real time PCR.
PE270 (IPPC0658) Comparing changes in defence-related gene expression upon root-knot nematode attack in susceptible and resistant rice cultivars Chanchal Kumari*, Tushar Kanti Dutta and Uma Rao Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Rice is one of the major staple food crops in the world and an excellent model system for studying monocotyledonous plants. Diseases caused by nematodes in rice are well documented, and among them root-knot nematode (RKN), Meloidogyne graminicola, causes extensive damage in yield. It is, therefore, necessary to identify novel sources of natural resistance to RKN in rice and to investigate the rice-RKN interaction in detail to understand the basal plant defence mechanisms and nematode manipulation of the host physiology. To this end, six different cultivars of rice were initially screened for RKN infection and development in pluronic gel medium, which resembles three-dimensional soil environment. Pusa 1121 was found to be most susceptible and Suraksha and Vandana were most tolerant to RKN infection. In order to investigate the role of major hormone-regulated plant defence pathways in compatible/incompatible rice-RKN interaction, some well-identified marker genes involved in each pathway were evaluated for their differential expression through qRT-PCR. Results suggested that, wound-responsive jasmonate pathway, WRKY transcription factors and PR-protein machinery are strongly induced in Vandana at early infection, while MAPK cascades, salicylate and jasmonate pathway-related defence genes are strongly elicited in Suraksha at later stage. However, a consistent down regulation of defence genes was recorded in Vandana during later stage of infection. As most information on the molecular interplay between plants and nematodes has been generated on dicotyledonous plants, the current study will strengthen our basic understanding of plant-nematode interaction in the monocot crops, which will aid in defining future strategies for best plant health measures.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 251 Session E : Biotic Stress Management and Interactions
PE271 (IPPC0727) Understanding the role of CIPK6 in plant defense response Atish Sardar* and Debasis Chattopadhyay National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Calcineurin B-like (CBL) protein family consists of proteins similar to both the regulatory subunit of calcineurin and the + neuronal Ca2 sensor in animals. Unlike Calmodulins, which interact with a large variety of target proteins, CBLs specifically target a family of protein kinases referred as CBL-interacting protein kinases (CIPKs). A database search of the Arabidopsis + thaliana genome sequence revealed 10 CBLs and 26 CIPKs homologues. Expression patterns of these Ca2 sensors and protein kinases suggest their diverse functions in different signaling processes, including light, hormone, sugar, and abiotic and biotic stress responses. So far, there is only one report in rice on the role of CBL-CIPK pairs in defense signaling. All the Arabidopsis CIPKs, so far studied, were shown to have roles in abiotic stresses such as high Na+ and low K+, cold and drought. From our laboratory, it was shown that CIPK6 is involved in growth and development along with abiotic stress tolerance of plants. Microarray studies showed that atcipk6 knock-down plants had altered expression of genes related to biotic stresses. Our primary investigation showed that cipk6 knock-out (cipk6) plants were less susceptible in response to hemibiotrophic pathogen Pseudomonas syringae pv tomato DC3000 (Pst DC3000) in comparison to the wild-type plants. The expression of several genes involved in the salicylic acid-mediated defense response pathway also justified and established this observation. Moreover, cipk6 plants were also more sensitive to bacterial elicitors.
PE272 (IPPC0739) Biochemical characterization of host and non-host herbivore induced proteinase inhibitors in leaves of black gram (Vigna mungo) Mariyamma Gujjarlapudi*, Vanka Swaroop, Vadthya Lokya and Kollipara Padmasree Department of Biotechnology and Bioinformatics, School of Life sciences, University of Hyderabad, Hyderabad-500046, India *Presenting author: [email protected]
Plants have evolved sophisticated defense responses through transcriptional activation of specific factors such as proteinase inhibitors (PIs) by a network of interconnecting signaling pathways in which oligosaccharides, peptide systemin, salicylic acid, jasmonic acid, ethylene, and reactive oxygen species (ROS) play an important role during various biotic and abiotic stresses. PIs are anti-metabolic proteins which interfere with the digestive process of insects. Black gram (Vigna mungo) contributes to about 10% of the national pulse production. It is a host plant to Helicoverpa armigera, a polyphagous pest and a non-host plant to Corcyra cephalonica. In the present study, black gram was chosen as a model plant to understand and characterize the PIs induced in leaves in response to infestation by host and non-host insects. The induction of PIs was estimated by measuring trypsin and chymotrypsin inhibitory units and SDS-gelatin-PAGE after infestation of leaves by larvae of H. armigera and C. cephalonica at different time intervals. Treatment of leaves with methyl jasmonate and wounding to leaves served as positive controls. Electrophoretic profile of induced PIs showed low molecular weight and possessed both trypsin and chymotrypsin inhibitory activity indicating the possibility of belonging to Bowman-Birk proteinase inhibitor family. Further, a significant increase in ROS and malondialdehyde content in leaves upon infestation with H. armigera and C. cephalonica, as well as wounding and methyl jasmonate treatment indicate the role of ROS and its cross talk with methyl jasmonates in the induction of PIs during both host and non-host insect attack.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 252 Session E : Biotic Stress Management and Interactions
PE273 (IPPC0750) Chitinase gene from Xenorhabdus nematophila imparts insect tolerance to plants Saquib Mahmood1*, Mukesh Kumar1, Punam Kumari1, Nirupama Banerjee2 and Neera Bhalla Sarin1 1Laboratory of Genetic Manipulation for Stress Alleviation and Value Addition in Plants, SLS, JNU, New Delhi-110067, India 2School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Plants express a wide variety of genes in response to plant pathogen or infection by pests. The best characterized genes belonging to PR protein family are those that encode the lytic enzymes such as chitinase, which play a vital role in plant defense against fungal pathogens due to their ability to hydrolyse the chitin, a major constituent of the lining of gut membrane of insects and the cell wall of a number of plant pathogenic fungi. Chitinase from different sources has been used as an important means of biological control for fungal diseases. In the present study, we have isolated a 1.9 kb of chitinase gene from Xenorhabdus nematophila, a Gram negative, entomopathogenic bacterium. The chitinase gene was cloned and a 76 kDa protein corresponding to the gene was expressed in E. coli. The purified chitinase protein was orally insecticidal to the major crop pest, Helicoverpa armigera when it was fed to the larvae of insects. The chitinase protein was also over expressed in tobacco plants via Agrobacterium mediated transformation. The transgenic lines were phenotypically normal and healthy. The insect assays showed high mortality of the larvae when fed on transgenic plant leaves over expressing the chitinase protein and also 60-70% reduction in leaves damage as compared to untransformed control plants. Our results demonstrate that the chitinase isolated from Xenorhabdus nematophila is orally toxic to Helicoverpa armigera and can prove to be a potent candidate for pest control in plants in future.
PE274 (IPPC0787) Exploitation of seed proteinase inhibitors as biopesticides and their biochemical characterization Kollipara Padmasree* Department of Biotechnology and Bioinformatics, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India *Presenting author: [email protected]
Proteinase inhibitors (PIs) are proteinaceous molecules which inhibit the catalytic activity of proteolytic enzymes. PIs are ubiquitously present in plant storage organs like seeds and tubers, since these are the vehicles for the propagation and survival of the species. They play a major role in regulating endogenous proteolysis, act as reserve storage of essential amino acids and expressed during biotic as well as abiotic stress tolerance. Seeds usually accumulate PIs during maturation and their concentration is varied between 1-10% of the total seed proteins. PIs may accumulate constitutively or induced after pathogen or herbivore attack. They are classified as serine, cysteine, aspartic and metallo PIs according to the type of proteases which they inhibit. According to the homology, molecular mass and cysteine content, serine PIs are further classified into Bowman-Birk inhibitor (BBI) type and Kunitz inhibitor families. The BBIs are small cysteine-rich proteins (6- 9 kDa) with two reactive sites typically against trypsin and chymotrypsin, while Kunitz inhibitors are ~20 kDa proteins, usually made up of two polypeptide chains linked with two disulphide bonds with single reactive site for trypsin or chymotrypsin. The PIs were known to inhibit digestive gut proteases and thereby affect the growth and development of insect pests. In the present study, the PIs from the seeds of three leguminous plants were examined for their potential as biopesticides against three lepidopteran insects Achaea janata, Spodoptera litura and Helicoverpa armigera. Further, the biochemical properties of the PIs with insecticidal activity would be discussed in detail.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 253 Session E : Biotic Stress Management and Interactions
PE275 (IPPC0815) The ZFD1 is a novel positive regulator of salicylic acid (SA) mediated defense in Arabidopsis thaliana Nidhi Singh* and Ashis Kumar Nandi School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Plants are capable of defending pathogen attack for immediate relief from the infection as well as to memorize the infection for inducing higher level of defense during subsequent infection. Based on the mode of recognition of pathogens, immune systems of plants are broadly classified as pattern triggered immunity (PTI) and effector triggered immunity (ETI). Plant hormone salicylic acid (SA) plays crucial roles in modulating plant defense. Transactivator protein NPR1 is considered as master regulator of SA mediated defense. Screening of NPR1 suppressor identified several genes that function as negative regulators of NPR1-independeny pathway. One such mutation identified previously in our lab is constitutive defense without growth defect 1 (cdd1). Through differential expression profiling of cdd1 and WT plants, we identified a novel plant defense regulator Zinc Finger protein for Defense (ZFD1). The loss-of-function mutant of ZFD1 is susceptible towards both virulent and avirulent pathogens. Extent of loss of resistance in zfd1 mutants is comparable to that of npr1 mutants. In agreement with the mutant phenotype, the ZFD1over-expression lines are hyper-resistant to bacterial pathogens and express PR genes at higher level. Expression of ZFD1 gene is induced by pathogen inoculation and exogenous SA application. Pathogen induced accumulation of SA and its glycosylated derivative SAG is largely unaffected in zfd1 mutant and ZFD1 over-expression lines. The zfd1npr1 double mutant plants were more susceptible than single mutants, zfd1 and npr1.The Y2H screening identified two defense regulators NHL3 and VDAC2. The results suggest that downstream to SA, ZFD1 positively influences NPR1-independent SA signaling.
PE276 (IPPC0835) Enhancement of glyphosate tolerance in tobacco by co-expressing CP4 EPSPS and glyhosate detoxifying enzymes (igrA: Glycine Oxidase) Amaranatha Vennapusa1*, Ramu S. Vemanna2, Chinta Sudhakar1 and M. Udayakumar3 1Department of Botany, SK University, Anantapur, Andhra Pradesh, India 2Plantbiology Division, The Samuel Roberts Noble foundation, Ardmore, OK, USA 3Department of Crop physiology, GKVK, UAS, Bangalore, Karnataka, India *Presenting author: [email protected]
Weeds are major biotic constraint for crops yield. The Glyphosate Resistance crops (GR crops) play a vital role in modern agriculture for weed control. The existing commercial GR crops are developed by either mutated or insensitive form of EPSPS. One of the disadvantages in these crops is glyphosate residues persist in the plant tissue, which is not desirable. Hence, expression of glyphosate degrading enzymes like mutated glycine oxidase (GO) and igrA (Increased glyphosate resistance Aldo keto reductase) are more relevant to achieve glyphosate tolerance besides decreasing residual levels. With this back ground in the present study co- expression of igrA: GO: CP4 EPSPS in tobacco plants has been studied for glyphosate bio efficacy and residual glyphosate levels by comparing with single gene (igrA, GO, EPSPS) and double gene (igrA: GO, igrA: EPSPS) expressing plants. The bio efficacy of transgenics plants assessed based on invitro regeneration of transgenic leaf explants on invitro glyphosate media and also by exposing leaf disc to different concentrations of glyphosate. The transgenics expressing either single or double and multiple genes showed improved tolerance to glyphosate. These events showed higher chlorophyll content, less membrane and oxidative damage compared to wild type. However the transgenics co-expressing igrA: Glycine oxidase: CP4EPSPS showed significantly enhanced glyphosate tolerance compared to other transgenic events. The efficacy of detoxifying enzymes was assessed by in vitro studies in crude leaf protein extracts. The efficacy of glyphosate degradation was more in tobacco transgenics co-expressing glyphosate degrading enzymes, compared to other transgenics and wild type.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 254 Session E : Biotic Stress Management and Interactions
PE277 (IPPC0841) Identification and expression analysis of a gene associated with defence responses to Sclerotium rolfsii in groundnut (Arachis hypogaea L.) Sujit Kumar Bishi1*, Mahesh Kumar Mahatma1 and Dinesh N. Vakharia2 1Directorate of Groundnut Research, Junagadh Agricultural University, Junagadh-362001, Gujarat, India 2Department of Biochemistry, Junagadh Agricultural University, Junagadh-362001, Gujarat, India *Presenting author: [email protected]
Groundnut (Arachis hypogaea L.), is an important legume crop and Sclerotium rolfsii is a major soil borne necrotrophic fungal pathogen causing stem rot leading to heavy economic losses worldwide. However the defense responses in groundnut to this pathogen are poorly understood so far. For implementation of successful molecular strategy to control stem rot, identification and characterization of important defense related gene(s) are required. Plants produce cell wall- associated polygalacturonase-inhibiting proteins (PGIPs); whose over expression improve the resistance to plant fungal and bacterial necrotrophs by inhibiting the polygalactoronases (PGs), the cell wall degrading enzyme produced by the pathogens. PGIPs are encoded by small gene families, identified in several plants including oil seed crops such as soybean, bean, and mustard; however no reports are available for groundnut. From an unidentified EST sequence (HO115743.1) derived from groundnut seed having sequence similarity with PGIP encoding gene (PGIP) of other crops, specific primers were designed. A partial but putative PGIP was isolated using cDNA as template prepared from stem rot infected groundnut leaf tissues and the sequence submitted to the NCBI database (Accession No. KP844637). The PGIP expression showed its higher induction in tolerant as compared to the susceptible groundnut genotypes upon S. rolfsii infection suggesting its possible role in stem rot stress tolerance in groundnut. This is identified gene will provide a basis for further cloning and functional validation of the members of PGIP family and also will aid in an understanding of the regulatory mechanism of groundnut tolerance to S. rolfsii.
PE278 (IPPC0914) Transient RNAi based gene silencing using CgCOM1 mediated by Agrobacterium confers disease resistance to chilli against anthracnose Nanditha Krishnan Vimalakumari*, Binod Kumar Mahto and Swatismita Dhar The Energy and Resources and Institute (TERI) University, Vasant Kunj, New Delhi-110070, India *Presenting author: [email protected] Chilli (Capsicum annuum L) is one of the most important crops in the tropics. A major constraint in chilli cultivation is the economic losses, often as high as 50%, resulting from anthracnose disease caused by Colletotrichum spp. The genus Capsicum is extremely recalcitrant to stable transformation, which results in poor efficiency and reproducibility. Transient transformation offers a viable alternative to stable transformation, as it is relatively easy, occur at high frequencies and helps in the quick analysis of transformants. We have standardized a highly efficient protocol for transient transformation in chilli (var: Pusa Jwala) mediated by Agrobacterium tumefaciens using pCAMBIA 2301 vector which harbours npt II and GUS genes. Maximum efficiency of transformation was obtained when microinjection, infiltration and application of vacuum were carried out sequentially to the leaf explants. Transient transformation was then successfully used to introduce RNAi vector having CgCOM1 gene, from Colletotrichum gloeosporioides, in chilli leaves. The transformed leaves were later challenged with the fungal pathogen. The transformed leaves showed differential resistance to Colletotrichum sp. challenge in comparison to wild-type leaves. The analyses of the transformants were carried out by phenotypic characterization, determining the hypersensitive response and by gene expression experiments. The present study provides a reliable tool for functional genomics by enabling rapid assessment of gene function prior to stable transformation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 255 Session E : Biotic Stress Management and Interactions
PE279 (IPPC0915) Modulation of host immunity and nutrient allocation by a biotrophic pathogen Divya Chandran* Regional Centre for Biotechnology, NCR Biotech Science Cluster, Faridabad-121001, Haryana, India *Presenting author: [email protected]
Obligate biotrophic pathogens like the powdery mildew fungi manipulate living plant hosts to acquire nutrients while limiting host defense responses. Powdery mildew is one of the most devastating diseases limiting productivity of food legumes in India and other developing countries. Chemical methods used to control the disease are neither economical nor environmentally sustainable. Therefore, there is a need to develop innovative strategies to combat this disease. To date, few powdery mildew resistant legume varieties have been identified, with the identity of the genes conferring resistance and the underlying molecular mechanisms not known. Our major goal is to identify and target novel legume host genes associated with defense that limit growth of the pathogen as well as host compatibility factors required for pathogen growth and reproduction. We envisage that targeting a combination of such factors would result in dramatically reduced pathogen proliferation and contribute to durable resistance that is less likely to be rapidly overcome by pathogen counter- evolution. To address our goal we are performing infection site-specific analyses via laser microdissection and next generation sequencing in the Medicago truncatula-Erysiphe pisi model pathosystem. We expect to uncover novel host components, cellular processes, and pathogen effectors impacting pathogen growth, previously hidden in whole-tissue global expression analyses. In parallel, we are studying the role of plant sugar transporters in mediating carbon partitioning at the host-pathogen interface, a process critical to both pathogen and host growth. Our long-term goal is to engineer durable powdery mildew resistance in food legumes of agronomic import.
PE280 (IPPC0918) Inositol phosphates and their role in regulation and trigger of effector triggered immunity Jewel Jameeta Noor, Hitika Gulabani* and Saikat bhattacharjee Regional Centre for Biotechnology, NCR Biotech Science Cluster, Faridabad-121001, Haryana, India *Presenting author: [email protected]
Plant immunity offers a classic example wherein each invaded cell is sophisticated and sufficiently equipped to possess mechanisms of surveillance, defense regulation and trigger only upon invasion with minimum side-effects. Although manifestation of most responses occur through step wise communication cascades of receivers and transducers, signalling in Effector-Triggered Immunity (ETI) is an exception. Signaling towards transcriptional reprogramming is regulated by intricate and interlinked connections between immune players / regulators at specific cellular locales. The resulting networks offer a robust and rapid responses characteristic of ETI. Several reports suggest that ETI responses impinge on hormonal signal transduction pathways. However, even after decades of research molecular processes underlying these signaling events remain largely unknown. As secondary messengers soluble Inositol-Phosphates (InsPs) influence mRNA export, apoptosis, and control of transcription. Inositol-modified lipids (phosphatidylinositols, PtdIns) determine architecture of most eukaryotic membranes. In our assays several tested mutants with unregulated immunity have differential transcripts for key inositol biosynthesis genes. In an effort to identify specific InsPs in ETI we are profiling their levels in inducible ETI systems and in mutants with altered defense capabilities. In parallel, we are trying to identify InsP–interacting proteins in plants. Molecular complexes of central immune regulators with resistance proteins (R) often occur at membranous localization likely involving PtdIns. Specific associations in this complex are perturbed by multiple unrelated effectors. We are further characterizing the this association to gain insights into how ETI is regulated from misfiring in absence of a pathogen threat and how effectors target these associations.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 256 Session E : Biotic Stress Management and Interactions
PE281 (IPPC0921) Effector-triggered immunity against a phytopathogenic bacterium: regulation, perturbation and signaling Saikat Bhattacharjee* Regional Centre for Biotechnology, NCR Biotech Science Cluster, Faridabad-121001, Haryana, India *Presenting author: [email protected]
Effector-triggered immune responses in plants are induced upon sensing specific effector activities (termed avirulence factors; Avr) on either a cognate resistance protein (R) directly or on a specific host target which the R protein guards. Although intended for suppressing defenses, in a resistant plant the same effector actions betray a pathogen presence by disrupting of protein-protein associations in a ‘resistasome’ complexes comprising of a negative modulator that regulate R proteins, and positive modulators that promote defense. Using Arabidopdis thaliana-Pseudomonas syringae pv tomato (DC3000) pathosystem our earlier work has identified one such regulatory association of immune players. Upon disruption activated R proteins and positive immune regulators that are resident of this complex signal through intricate cross-linked networks that cause massive transcriptional reprogramming that culminate pathogen threat. Whereas many newer resistance specificities are being identified, detailed molecular mechanism of how immune assemblies regulate defenses in an un-invaded plant and how perturbations upon pathogen attack effectively couple to downstream signaling routes remains unknown. To gain further insights into these processes we are characterizing interactions that aid the assembly of a resistasome complex. Our analysis have identified that extensive post-translational modifications define strategic deployment of defense regulators and a phytopathogenic effector often impinge on and alter these modifications through a competing/interfering post-translational activity. Interestingly the identification of unrelated effectors targeting a common target further support this model. Our studies also propose that production of secondary metabolites that signal immune pathways are also effectively coupled to changes in protein modifications thereby ensuing a robust link between trigger and signaling during plant innate immunity.
PE282 (IPPC0922) How SWEET is biotrophic life? Role of plant sugar transporters in pathogen nutrition and plant defense Megha Gupta* and Divya Chandran Regional Centre for Biotechnology, NCR Biotech Science Cluster, Faridabad-121001, Haryana, India *Presenting author: [email protected]
Obligate biotrophic pathogens like the powdery mildew fungi modulate plant host metabolism to divert nutrients, especially sugars, to fuel their own growth at the expense of plant productivity. They form specialized feeding structures (haustoria) within host epidermal cells and create an apoplastic interface through which host sugars, released primarily through the action of sugar transporters, are absorbed. Recently, a new family of plant sugar efflux transporters, known as SWEET, has been implicated in bacterial and fungal nutrition. For example, the bacterial blight pathogen Xanthomonas oryzae secretes transcription-activator like (TAL) effectors that directly induce the expression of specific SWEET genes triggering sugar efflux at the host-pathogen interface. Notably, when SWEET expression is blocked, so is the growth and virulence of the pathogen. To ascertain the role of SWEET transporters in powdery mildew nutrition we utilized the Medicago truncatula (legume host)-Erysiphe pisi (pathogen) interaction as a model pathosystem. Expression levels of 24 MtSWEET candidates (identified through bioinformatics) were quantified via real-time quantitative PCR at various time points post E. pisi inoculation. We found that a number of MtSWEETs exhibited enhanced expression in response to E. pisi in a susceptible host with peak expression observed just prior to the onset of fungal conidiation. In contrast, E. pisi-induced expression was either absent or reduced in a resistant host with peak expression detected at earlier time points coinciding with fungal growth arrest. Our results highlight the possibility of a dual role for plant SWEET transporters in powdery mildew nutrition as well as plant defense.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 257 Session E : Biotic Stress Management and Interactions
PE283 (IPPC0929) The ‘Trigger’ in effector-triggered immunity: deregulation via post-translational modification Kishor Ingole*, Bornali Gohain and Saikat Bhattacharjee Regional Centre for Biotechnology, NCR Biotech Science Cluster, Faridabad-121001, Haryana, India *Presenting author: [email protected]
Basal immunity (also called PAMP-triggered immunity, PTI) in plants, induced against conserved microbe determinants, is suppressed by adapted pathogens that secrete effectors and interfere with specific host defense signaling routes (termed virulence). As a counter-defense strategy resistant plants deploy specific resistant (R) proteins that either directly or indirectly sense the effector-induced modifications and mount effector-triggered immunity (ETI). Pathogens that trigger ETI are termed as avirulent. The outcome of ETI is a massive transcriptional reprogramming that culminates pathogen colonization and spread. In many instances unrelated effectors target common virulence target. The specific R protein that gets activated depends on the nature of post-translational modifications (PTMs) an effector causes on target. PTM by SUMOylation regulate key processes in normal growth and development such as protein stability, localization, and protein– protein interactions. Transcriptional regulation is also achieved through SUMO-dependent modification of histones. In plant defenses although SUMO and SUMOylation machinery components are implicated, their precise role remains unexplored. We are using mutants with altered defense responses to gain insights into specific roles of individual SUMO isoforms in plant defenses. Our preliminary observations suggest induction of a specific Arabidopsis SUMO member upon effector-triggered immunity. We also observe global upregulation of SUMOylated proteins in plants with uncontrolled defenses. We are focusing on further characterizing SUMO functions and identifying SUMOylated targets and elucidating their roles in ETI.
PE284 (IPPC0939) Functional characterization of a key regulator of innate immunity, CaRACK1, in chickpea in response to necrotrophic fungus Ascochyta rabiei Sandhya Verma*, Kamal Kumar and Praveen K. Verma Plant Immunity Laboratory, NIPGR, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Plants are constantly under threat of infection by myriads of potential pathogens. In order to combat pathogen attack, plants have developed various intricate mechanisms that results in the establishment of effective defense system. A first layer of this system is based on perception of PAMPs through pattern recognition receptors (PRRs) at the plant’s cell surface. Successful pathogens produce effectors to inhibit innate immunity, but plants, in turn, can perceive such effectors through additional receptors to mount induced immunity. Necrotrophic plant pathogenic fungi possess a battery of lytic and degradative enzymes. Moreover, recent studies have shown that they can also subtly manipulate the host defense during infection in a manner similar to biotrophic pathogens. In chickpea, the necrotrophic fungus Ascochyta rabiei causes blight disease. Our interest is to study signaling mechanisms required for induced resistance during pathogen interaction in chickpea. Till date, the role of signaling components against fungi has remained elusive. RACK1 (Receptor for Activated C-Kinase 1), that has been isolated in our laboratory, functions as a scaffold protein by physically interacting with many other proteins and facilitating their interactions. During chickpea-Ascochyta infection, CaRACK1 is highly expressed suggesting its role in plant defense. Moreover, this conserved WD40-repeat-containing protein exists in multi-meric form in vitro. Functional complementation of CaRACK1 in yeast ASC1 mutant rescued the yeast mutant phenotype. It suggests CaRACK1 is a functional homolog of S. cereviseae ASC1. The importance of CaRACK1 in various key signaling events also suggests that CaRACK1 might be a potential target of Ascochyta rabiei effectors.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 258 Session E : Biotic Stress Management and Interactions
PE285 (IPPC0974) Integrated analysis of miRNA and mRNA omics data from diseased tomato plants: possible application in stress management Deepti Sarkar, Jayanti Jodder, Sayani Dey, Ranjan Kumar Maji, Rohit Das, Arunava Mandal, Payel Bhattacharjee, Arijita Sarkar, Zhumur Ghosh, Pallob Kundu* Division of Plant Biology and Bioinformatics Center, Bose Institute, P1/12 CIT Scheme VII M, Kolkata, West Bengal, 700054, India *Presenting author: [email protected]
Gene expression cascade in a plant is altered in times of stress. Reprogramming of the expression profiles of genes are required for a robust and specific response. MicroRNA-mediated regulation of gene expression plays a significant role in this process as a miRNA is capable of regulating expression of multiple genes both at post transcriptional and translational level. Until now, tomato miRNAs responsive to Alternaria fungal stress were not known. We will present profile of miRNAs that are differentially expressed during Alternaria stress in local tomato cultivar (Pusa Ruby) and results of integrating the expression profile with transcriptome analysis of tomato. The knowledge gathered from these omics studies has been applied to get an insight into the miRNA-mediated regulation of ‘disease-resistance’ genes in tomato. Each miRNA plays multiple roles, thus deliberate manipulation of their expression profiles for disease management would require thorough knowledge about the regulation of their biogenesis. We will also present evidence of complex regulation of a miR-gene expression and suggest how its expression can be manipulated using genome manipulation tools.
PE286 (IPPC0994) Control of sugarcane mosaic disease using bacterially expressed viral coat protein specific double strand RNA - A RNA interference approach Yasir Umar Pattiwala* and Prabu Gajjeraman Department of Biotechnology, Karpagam Academy of Higher Education, Coimbatore-641021, Tamil Nadu, India *Presenting author: [email protected]
Sugarcane production has declined in recent years due to several adverse factors, including mosaic viral disease. Sugarcane Streak Mosaic Virus (SCSMV) is a major cause for mosaic disease alaongwith Sugarcane Mosaic Virus (SMV) and Sorghum Mosaic Virus (SrMV). RNA silencing plays a significant role in plant resistance against viruses, transposons and other microbes. The phenomenon of RNA interference (RNAi) is involved on sequence-specific gene regulation driven by the introduction of dsRNA resulting in inhibition of transcriptional or translational suppression. Coat protein (CP) encoded gene of Sugarcane Streak Mosaic Virus (SCSMV) was amplified in sense and anti-sense orientation and cloned into RNAi cloning vector to produce double strand RNA (dsRNA). The culture medium and conditions were optimized for high level bacterial expression of SCSMV_CP dsRNA. The sugarcane sett treatment method was optimized to induce SCSMV_CP dsRNA mediated RNA interference and resulted 70% reduction of virus titer than the untreated counterparts. These results provides a valuable means for controlling plant viral disease using bacterially expressed dsRNA and RNAi approach.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 259 Session E : Biotic Stress Management and Interactions
PE287 (IPPC1045) Biochemical changes in different cultivars of Aonla (Emblica officinalis G.) during storage at room temperature Jitender Kumar*, Poonam Kumari and Suman Bala Department of Botany and Plant Physiology, CCS Haryana Agricultural University, Hisar *Corresponding author: [email protected]
An experiment was conducted to study the biochemical changes in five cultivars of Aonla during storage at room temperature. Acidity content of fruits decreased during storage from 3rd to 18th day of storage in all the cultivars. More acidity content was observed in the fruits of cv. Krishna (1.825%) after 18th day of storage. Whereas minimum acidity was retained in the fruits of cv. Chakaiya (1.574%) after same period of storage. Ascorbic acid content of fruits also decreased with increasing period of storage in all the cultivars of aonla fruit. Maximum decrease was observed in the fruits of cv. Krishna (441.2%) after 18 days of storage whereas minimum decrease was observed in Chakaiya fruit (476.7%). TSS content of fruits also decreased with increasing period of storage in all the cultivars. Minimum TSS was retained in the fruits of Banarasi (7.70Brix) whereas maximum TSS was retained in Hathijhul fruits (90Brix).
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 260 Session F : Plant Nutrition Physiology and Genetics
PF288 (IPPC0012) Physiological characteristics and yield improvement by Arbuscular mycorrhizal fungus in wheat (Triticum aestivum L.) Kamal Dutt Sharma1*, Karmal Singh1, Rajesh Gera2 and Jagdev Singh1 1Crop Physiology Lab, Department of Agronomy, CCS Haryana Agricultural University, Hisar-125004, Haryana 2Department of Microbiology, CCS Haryana Agricultural University, Hisar-125004, Haryana *Presenting author:[email protected]
Arbuscular mycorrhizal (AM) fungi are ubiquitous root symbionts of more than 90% of vascular plants and are thought to contribute in plant nutrition, particularly phosphorus. A field experiment was conducted in split plot design, with wheat genotypes, WH 1021, WH 1105, WH 1123, WH 1124, WH 1158 in main plots and seed treatments practices, i.e., dry seeding, primed seed (seed soaking overnight in water), primed seed + AM fungi (Glomus mosseae), primed seed + Azotobacter in the sub plots with three replications under late sown situation. Physiological characters of flag leaves were measured after anthesis. Wheat genotypes sown with water primed, seed inoculated with AM fungi and Azotobector increased grain yields by 2.4, 8.1 and 3.8%, respectively over conventional practice (3998 kg ha-1) under late sown condition. The increase in gain yield was mainly due to early seedling emergence and vigour, more number of spikes per plant and higher biomass and improved physiological traits like higher membrane stability and photosynthetic efficiency of flag leaf during anthesis. Among the genotypes, WH 1105, followed by WH 1158 showed significantly higher yield (4315 and 4278 kg ha-1) over the other tested genotypes due to higher membrane stability, more tillering and test weight. The increase in seed yield by seed priming and AM fungi was positively associated with days to emergence and heading, membrane stability and photosynthetic rate.
PF289 (IPPC0142) Agriculture and nutritional security: Enhancement of lysine contents in rice through genetic manipulation of aspartate metabolic pathway enzymes Manjari Mishra1*, Sneh L. Singla Pareek2 and Ashwani Pareek 1Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India 2Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Malnutrition is a major problem in developing countries of the world. Due to lack of nutritious food, more than one third of children are dying every year, specially in developing countries. This scenario impose challenges to improve nutritional status and nutritional security in economically poor regions of the world. Inspite of being a staple food for a large part of world’s human population, rice is nutritionally poor for human and animal consumption because the proteins are deficient in certain essential amino acids, especially, lysine. A complex and tightly regulated metabolic pathway is involved in lysine synthesis in plants where the two enzymes DHDPS (Dihydrodipicolinate Synthase) and LKR/SDH (Lysine Ketoglutarate Reductase / Saccharopine Dehydrogenase) tightly regulate lysine biosynthesis and degradation, respectively. In the present work, we have genetically manipulated the lysine pathway enzymes towards metabolic engineering of lysine content in rice. We have generated overexpression construct, RNAi construct and lysine double construct and generated transgenic plants of Oryza sativa cv. IR-64. We got maximum increase in lysine content in Lysine Double Construct (LDC) lines i.e. 58% compared to the wild type. These transgenic lines will be further used for metabolic profiling to assess the effects of increased lysine content on other metabolites.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 261 Session F : Plant Nutrition Physiology and Genetics
PF290 (IPPC0152) Changes in growth, yield, juice quality and biochemical attributes of sugarcane in response to silica granules Anshu Singh1*, Smita Singh1, Neeru Jain2, P. Lal1, R.K Singh1, Asha Gaur1, S.K Shukla1, S.P. Singh1, A. Chandra1 and Radha Jain1 1Indian Institute of Sugarcane Research, Lucknow-226002, India 2Privi Life Sciences Pvt. Ltd. Navi Mumbai, India *Presenting author: [email protected]
Silicon (Si), the second most abundant element in the earth crust (27.7%) is usually found in the form of complex silicate minerals, and less often as silicon dioxide. It plays an important role in inducing resistance to various biotic and abiotic stresses in plants, helps in controlling Al, Mn and Fe toxicities, increases P availability, reduces lodging and improves rate of photosynthesis by effective use of sunlight as well as efficient management of plant water economy. Sugarcane, a typical Si- accumulating plant is known to absorb a large amount of silica from the soil. A field experiment was conducted to study the effect of Silica granules (obtained from Privi Life Sciences Pvt. Ltd., India) on growth, yield and juice quality attributes of sugarcane at IISR Experimental Farm, Lucknow during 2013-15 (autumn season). The treatments comprised of control (T1), silica granules @ 20 (T2), 40 (T3) and 80 kg/ha (T4), and calcium silicate too high (T5). Specific leaf weight, total dry matter and nitrate reductase activity increased in all treatments relative to control. Silica granules @ 40 kg/ ha (T3) showed highest increase in yield attributes; cane length, girth, NMC and cane yield. oBrix, juice purity, sucrose % juice, juice extraction, CCS per cent juice, S/R ratio and SPS were comparatively higher in treated plants. Highest soluble silica content was observed in T2 (leaf) and T5 (root) treatment. Findings obtained suggested that application of silica granules holds immense potential to improve yield and juice quality attributes of sugar cane.
PF291 (IPPC0158) Calcium regulates tuberization in potato through enhanced expression of CaM, StCDPK and LOX genes Devanshi Chandel Upadhyaya*, Abhishek Kumar, Deepak Singh Bagri and Chandrama Prakash Upadhyaya Department of Biotechnology, Dr Harisingh Gour Central University, Sagar-470003, M.P., India *Presenting author: [email protected]
Calcium plays an important role in plant physiology and various plant cell signaling pathways, and also alters many biochemical processes by activating specific enzymes. Cytosolic Ca2+ regulates the activity of Ca2+ dependant protein kinases, which phosphorylate various key metabolic enzymes. In the present study, expression of two Ca2+ dependant proteins, calmodulin (CaM1) and calcium dependant protein kinase (StCDPK), as well as the lipoxygenase (LOX; EC 1.13.11.12) were studied upon Ca2+ application to the single node segments of potato inoculated for tuberization. Calcium at higher levels (6 mM) significantly improved the tuber number, tuber growth and tuber yield under in vitro conditions. The expression levels of the CaM1 and StCDPK were significantly higher in stolons showing positive correlation with supplemental Ca2+ and tuberization response. Similar trends were observed with LOX enzyme activity, which increased by 18% and 25% with the addition of Ca2+ at 3 and 6 mM, respectively to the tuber induction medium, when compared to control. Thus, present study reports that the increase in tuberization, tuber growth and tuber yield with the supplementation of Ca2+ could be attributed to the increased expression of the Ca2+ dependant proteins and enhanced lipoxygenase activity.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 262 Session F : Plant Nutrition Physiology and Genetics
PF292 (IPPC0223) Influence of silicon solubilizers on yield and biotic stress tolerance in rice (Oryza sativa L.) genotypes Babita Joshi, Deepti Shankhdhar, Alok Shukla and Shailesh Chandra Shankhdhar* Department of Plant Physiology, College of Basic Sciences & Humanities, G.B. Pant University of Agriculture & Technology, Pantnagar- 263145, Uttarakhand, India *Presenting author: [email protected]
A field experiment was conducted in kharif season of 2014 to investigate the effect of silicon solubilizer on different rice genotypes namely DRRH-3, PA-6129, PA-6201, PA-6444, PHB-71 and BPT-5204. Soil application of solid silicon solubilizer (imidazole) was given at the time of maximum tillering and panicle initiation. Various growth, biochemical parameters and yield attributes were observed. Insect infestations were also recorded at two growth stages (panicle initiation and flowering). The leaf area index (LAI) was increased 27.55% as compared to control at maximum tillering, 7.05% increased at panicle initiation and 16.67% increased at flowering in DRRH-3. As compared to control 5.54% increase in economic yield was obtained in PA-6129 however 5.65% increase in harvest index was found in case of DRRH-3. Silicon solubilizers had a significant effect on iron content in different rice genotypes. Iron content was increased by 73.36% in grains of BPT-5204 with husk and 78.50% without husk in PA-6201. The zinc content was increased by 52.61% in grains of DRRH-3 with husk and 29.82% without husk in grains of PA-6129. Pest infestations were also significantly reduced in all genotypes of rice as compared to control. In BPT-5204, 60.0% decreased in dead heart infestation and 63.64% decreased in white ear head infestation was observed in PA-6201 as compared to control. Amongst the six genotypes PA-6444 was the most efficient in utilizing silicon under soil application of imidazole carrier molecule.
PF293 (IPPC0228) Functional characterization of OsPAP9 under P deficiency Bipin Pandey*, Poonam Mehra and Jitender Giri National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Phosphorus (P) is an essential macronutrient for plant growth and development. Most of the soil P exists in bound form as insoluble complexes, not readily available to the root system. Thus, P is one of the most limiting nutrients for plant growth. However, plants have evolved a set of adaptive morphological, physiological, biochemical and molecular responses that allow them to withstand low P stress in soils. Among these, intracellular purple acid phosphatases (PAPs) play important role in P recycling from cellular P reserves whereas extracellular PAPs are largely secreted into the rhizosphere to scavenge P from bound organic sources. In rice, 26 PAPs have been identified, out of which we found ten PAPs being significantly expressed under P deficiency. However, except OsPAP10a, no detailed study of any of the rice PAPs has been carried out so far. Therefore, we selected one of the differentially expressed PAPs, OsPAP9 to improve low P tolerance. Biochemical studies of OsPAP9 revealed that it acts as functional phosphatase at acidic pH and is highly thermostable. Interestingly, OsPAP9 showed wide spectrum of substrate specificity. Moreover, OsPAP9 requires a divalent cation Mg2++ as best cofactor for its optimal activity. Constitutive overexpression of OsPAP9 showed improved growth performance of rice plants as compared to WT under low P conditions. Interestingly, growth of overexpressed lines was relatively vigorous when low P starved plants were supplemented with ATP as organic P source. Thus, OsPAP9 transgenic rice will be a useful resource for alleviating P deficiency.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 263 Session F : Plant Nutrition Physiology and Genetics
PF294 (IPPC0241) Enhancing flavour and quality of tomatoes by expression of synthetic gene encoding sweet protein monellin Tanushri Kaul* and Chinreddy Subramanyam Reddy Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi-110067, India *Presenting author: [email protected]
Monellin a sweet-tasting protein exists naturally as a heterodimer of two non-covalently linked subunits chain A and B and it drops sweetness on denaturation. In this study, we validated the expression of a synthetic monellin gene encoding a single polypeptide chain covalently linking the two subunits under T7 and fruit-ripening specific promoters in E. coli and tomato fruits, respectively. Purified recombinant monellin protein retained its sweet flavor at 70 C and pH 2. We developed 15 transgenic T0 tomato plants overexpressing monellin that were devoid of any growth penalty or phenotypic abnormalities during greenhouse conditions. T-DNA integration and fruit-specific heterologous expression of monellin had occurred in these transgenic tomato lines. ELISA revealed expression of monellin was 4.5% of total soluble fruit protein. Functional analyses of transgenic tomatoes of T2-5 and T2-14 lines revealed distinctly strong sweetness as compared to wild type. Monellin a potential non-carbohydrate sweetener if expressed in high amounts in fruits and vegetables would enhance their flavour and quality.
PF295 (IPPC0287) Leaf and grain nitrogen partitioning of a dual-adapted rice variety grown under transplanted and directed seeded condition A. Amaregouda*, B.O. Kiran, Supriya Patil, Benoit Clerget and B.K. Desai Deparment of Crop Physiology, University of Agricultural Sciences, Raichur-584104, Karnataka, India *Presenting author: [email protected]
Fertilizer management practices need to be developed for rice production, a system in which rice is grown under transplanted and direct seeded rice cultivation. We evaluated the effects of graded nitrogen (N) levels application on the yield of transplanted and direct seeded rice under rainfed conditions in 2013 wet seasons at Main Agricultural Research Station, Raichur state, India. Yields increased with increasing N rate from 75-125% RDN (Recommended Dose of Nitrogen). Better translocation of N was observed with 185 kg N ha-1 as compared to 150 kg N ha-1 in both the methods of crop establishment. The grain N content was higher (1.52%) in JKPH 3333 under direct seeded condition at 185 kg N ha-1. At harvest significantly higher leaf nitrogen content (1.01%) was recorded in direct seeded rice compared to that of transplanted rice (0.66%). Although the yields were higher in transplanted rice, well developed root system enhanced better allocation of N at harvest in direct seeded rice cultivation. However, among different planting methods, significantly higher nitrogen content (0.89%) was recorded in transplanted rice as compared to that of direct seeded rice (0.54%). Cultivation of rice with direct seeded method (42.92 q/ha) resulted in 33.33% yield reduction compared with transplanted method (56.29 q/ ha). Among the yield attributes, number of spikelets/panicle was significantly affected under aerobic method, while test weight and harvest index were not influenced by the method of water management. The traditional method of rice cultivation resulted in higher total biomass production as compared to direct seeded rice cultivation. Our result suggested that that selection for JKPH 3333 N accumulation may have value in variety for nitrogen responsive and stable yields even under water deficit conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 264 Session F : Plant Nutrition Physiology and Genetics
PF296 (IPPC0307) Arbuscular mycorrhiza (Rhizophagus irregularis) influences As accumulation, growth and productivity in Cajanus cajan (L.) Millsp. (pigeonpea) genotypes exposed to AsIII and AsV stress Lakita Kashyap* and Neera Garg Department of Botany, Panjab University, Chandigarh-160014, India *Presenting author: [email protected]
As exists naturally in the form of arsenate (AsV) and arsenite (AsIII) in soil which affect crop growth and productivity. Arbuscular mycorrhizal (AM) associations impart abiotic stress tolerance by improving growth and nutrient status (P). However, reports on the relative role of AM in reducing AsIII and AsV toxicities in legumes are scanty. Thus, experiments were conducted to ascertain the modulatory role of AM in alleviating arsenate (AsV) (25 mg/kg and 50 mg/kg) and arsenite (AsIII) (5 mg/kg and 10 mg/kg) induced responses in terms of growth, water relations and productivity in pigeonpea genotypes [tolerant Pusa 2002 and sensitive Pusa 991]. Results indicated that at higher concentrations, total As was detected in roots, leaves and seeds in AsIII treated plants while in roots and leaves in case of AsV. Moreover, Pusa 991 accumulated higher As than Pusa 2002. A significant decline in mycorrhizal colonization (MC), chlorophyll a/b ratio, RWC, root to shoot ratio and ultimate productivity was recorded under both As species, with AsIII more toxic than AsV. However, AM reduced total As accumulation in both AsV and AsIII treated plants which resulted in significant improvement in plant biomass, reduced pigments degeneration and increased ultimate yield. Pusa 2002 was more responsive to AM, established more effective MC and was found to be significantly more tolerant to both AsV and AsIII stresses when compared with Pusa 991. Study indicated that although R. irregularis alleviated both AsV and AsIII toxicities, percent benefits of AM were relatively more significant in case of AsIII treated plants of pigeonpea.
PF297 (IPPC0308) Cumulative effects of Silicon supplementation and mycorrhization in restoring growth, yield and ionic homeostasis of chickpea genotypes subjected to long term salinity Purnima Bhandari * and Neera Garg Department of Botany, Panjab University, Chandigarh-160014, India *Presenting author: [email protected]
Silicon (Si) uptake varies among the plant species and has been reported to be low in legumes (<0.5%). Both, Si amendment to the soil and arbuscular mycorrhizal (AM) symbiosis stimulate plant tolerance to salinity. Few reports indicate that AM fungi increase Si uptake in mycorrhizal plants. However, reports on the comparative role of exogenous application of Si and AM inoculations, as well as their interactive effects through combined treatments, in improving plant resilience to salt stress are lacking. Therefore, pot trials were undertaken to ascertain the influence of AM fungal species - Funneliformis + + + mosseae and/or Si supplementation (@ 4mM) on Na uptake and translocation, nutrients acquisition (N, P, K , Ca2 ) and resultant yield in salt tolerant (HC 3) and sensitive (CSG 9505) Cicer arietinum L. (chickpea) genotypes under salt stress (0-100 mM). Results indicated that although both, Si and mycorrhiza improved plant biomass and enhanced yield under stressed conditions, Si-mediated benefits were elicited through significantly reduced Na+ uptake, coupled with enhanced + + + + + + K , Ca2 contents thereby, restoring K /Na , Ca2 /Na ratios, while mycorrhizal effects were directed towards improved plant growth, enhanced N, P acquisition and the resultant seed yield. Interestingly, under combined treatments, the benefits achieved were substantially higher than the individual Si as well as AM applications, suggesting that mycorrhiza- mediated Si absorption from the soil contributed towards enhanced salt tolerance and seed yield, more in HC 3 than CSG 9505 genotypes. Results strongly highlighted the usage of combined Si-mycorrhization as an effectual strategy in increasing chickpea productivity in salt contaminated areas.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 265 Session F : Plant Nutrition Physiology and Genetics
PF298 (IPPC0309) Silicon nutrition and Arbuscular mycorrhizal (AM) inoculation as an effective strategy to improve growth and nitrogen fixing efficiency of Cajanus cajan (pigeonpea) genotypes under cadmium stress Sandeep Singh * and Neera Garg Department of Botany, Panjab University, Chandigarh-160014, India *Presenting author: [email protected]
Cadmium (Cd) is amongst the most hazardous elements and its presence at elevated levels in the soils is threatening agricultural productivity. Role of silicon (Si) nutrition and AM in mitigating Cd stress has gained importance in recent years. The present study was aimed to evaluate individual and cumulative effects of Si (600 mg/kg) and AM (Rhizophagus intraradices) on Cd (CdSO4 – 25, 50 mg/kg) induced responses in pigeonpea genotypes (Pusa 2002 and Pusa 991). Cd stress caused significant reduction in dry matter, with Pusa 991 exhibiting higher decline than Pusa 2002. The impact of Cd was more pronounced in roots than shoots (decreased R/S ratio) and this decline was proportionate to Cd uptake and translocation. Decreased root biomass reduced nodulation potential and ultimate N accumulation under metal stress. Individual applications of both Si and AM fungal inoculations counteracted the negative effects of Cd stress and significantly improved growth and nitrogen fixing potential, with higher positive effects in Pusa 2002 than Pusa 991. AM was highly efficient in reducing Cd uptake thereby improving root biomass and N fixing efficiency (ARA) while Si was more effective in improving shoot biomass. Combined application of Si and AM ameliorated the negative effects of Cd25 with significant improvement under Cd50. Pusa 2002 was more efficient in establishing mycorrhizal colonization, Si uptake and was more tolerant to Cd stress than Pusa 991. Thus, the study suggested that combined treatments of Si and AM can be used as an effective strategy in combating Cd stress in pigeonpea.
PF299 (IPPC0321) Over expression of MusamiR398a in transgenic banana plants towards improving the iron content Prashanti Devendra Patel* and Thumballi Ganapathi Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumabi-400085, Maharashtra, India *Presenting author: [email protected]
Banana, an important food crop, though macronutrient-rich, is poor in iron. Hence it is imperative to understand the different mechanisms of Fe homeostasis in the plant so that banana can be enriched in iron to alleviate anaemia. Thus, the present study was undertaken to understand the roles of micro RNAs in iron homeostasis. The plant miRNAs 397, 398 and 408 are well-known copper-miRNAs that target various Cu- proteins under Cu deficiency. This diverts the precious metal to the essential Cu protein plastocyanin that can only use copper as a cofactor. However these miRNAs were also down regulated under Fe deficiency, in common bean and Arabidopsis, suggesting probable roles in the cross talk between Fe and Cu homeostasis. In this study, one of the two loci of banana miRNA 398- miR398a was retrieved from the Banana Genome Hub after homology searches. After sequencing, the stem loop sequence along with flanking nucleotides was cloned into the pCAMBIA 1301 binary vector, under the constitutive ZmUbi promoter. The plant expression vector was electroporated into Agrobacterium tumefaciens strain EHA 105 and used to transform banana embryogenic cells. The transgenic plants were regenerated on hygromycin selection medium. Initial screening of plants for transgenic nature, was done by GUS histochemical staining and subsequently the plants were hardened in the greenhouse. Further analysis by PCR using ZmUbi and gene specific primers was carried out to confirm their transgenic nature. The analysis of these transgenic plants would help understand how this miRNA influences iron homeostasis in banana.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 266 Session F : Plant Nutrition Physiology and Genetics
PF300 (IPPC0326) Evaluation of organic and inorganic fertilizers on neutral volatile aroma compounds of burley tobacco Siva Raju Kemisetti*, D. Damodar Reddy and P. Harishukumar Central Tobacco Research Institute, Rajahmundry-533105, Andhra Pradesh, India *Presenting author: [email protected]
A field experiment was conducted during 2011-13 at Burley Tobacco Research Station, Kalavacherla, East Godavari district, to study the effect of organic and inorganic manures with three formulations of organic manures (Neem cake, Pongamia cake, filter press cake, poultry manure, FYM and vermi compost in different ratios) and two levels of inorganic fertilizers (120 and 160 kg N/ha) on neutral volatile aroma compounds (NVAC) of burley tobacco. Cured leaf samples of top leaf position from different treatments were analysed for NVAC by GC-MS. The NVAC detected includes phenylalanine metabolites, carotenoids, thunberganoids, terpenoids and neophytadiene. Among the terpenoids, hydroxysolavetivone, solvitevone, caryophyllene oxide, nerolidiol epoxiacetate, phytol, globulol and epiglobulol are the major constituents whereas megastigmatrienone and dihydroactinidiolide are major constituents in carotenoid degradation compounds. As shown by their relative content in NVAC fractions, thunberganoids and cembrenoids content was maximum (310 to 505 ppm) followed by terpenoids (255 to347 ppm), neophytadiene (103 to 156 ppm), carotenoids (65 to 90 ppm) and phenylalanine compounds (11.14 to 20.82 ppm). The five isomers of megastigmatrienone were present in all the treatments except 160 kg N/ha and megastigmatrienone isomer-2 was in maximum content. Megastigmatrienone isomers content was significantly higher in manure C and B treatments when compared to inorganic treatments. With increase in nitrogen level from 120 to 160 kg N/ha, phenylalanine compounds, terpenoids, carotenoids, neophytadiene and thunberganoids contents were decreased by 41.81, 10.21, 8.14, 17.6 and 35.8% respectively. The results revealed that increased level of inorganic nitrogen significantly decreased the NVAC and application of 25% of nitrogen in the form of organic manure along with the inorganic nitrogen to the burley tobacco significantly increased the NVAC as compared to application of inorganic nitrogen alone.
PF301 (IPPC0349) Overexpression of Musa ferritin gene in transgenic banana plants for increasing the iron content Karuna Yadav* and Thumballi Ganapathi Plant Cell Culture Tech. Section, Nuclear Agric. & Biotechnology Division Bhabha Atomic Research Centre, Mumbai-400085, India *Presenting author: [email protected]
Banana is one of the most important fruit crop and is a cheap source of nutrients and vitamins. However, the fruit is deficient in the micronutrient iron and fails to meet the expected daily dietary requirement as per FDA. The iron deficiency manifests as anaemia in deprived sections of society. In the present study overexpression of native iron storage protein ferritin is undertaken to enhance the iron content in banana plants. Ferritin, an iron storage protein with a capacity to sequester up to 4500 Fe atoms, protects the plant from iron induced oxidative stress. Homology search in banana using rice sequences predicted two sequences in EST database. These two genes have been amplified using cDNA from banana leaves. The full length amplified fragments were cloned into pCAMBIA1301 plant expression vector under ZmUbi constitutive promoter using Pst1 and Kpn1 restriction sites. Agrobacterium tumefaciens mediated transformation of banana embryogenic cells was carried out. The transformed somatic embryos developed on selection medium containing hygromycin. Putatively transformed regenerated lines were identified by GUS histochemical staining. Genomic DNA PCR analysis for these lines was carried out to confirm the transgenic nature of the regenerated plants. A total of 22 transgenic lines were regenerated. Copies of each transformed lines are maintained on banana multiplication medium as multiple shoot cultures. One clonal copy was then rooted on MS media supplemented with NAA and hardened in the green house for further analysis. The results from this study would help in improving the iron content in transgenic banana plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 267 Session F : Plant Nutrition Physiology and Genetics
PF302 (IPPC0352) In silico analysis of EF-hand domain containing protein in five different plant species and analyse of competitive binding of divalent and monovalent ions for metal binding site Aditya Nath Jha, Tripathi Kalyani and Ram Singh Purty * University School of Biotechnology, Guru Gobind Singh Indraprastha University, Sector 16C, Dwarka, New Delhi-110078, India *Presenting author: [email protected]
Metallo-proteins are the proteins that contain a metal ion as a cofactor. Calcium binding proteins are very commonly found metallo-proteins, which play very significant role in signaling pathways, cell cycle and various metabolic pathways. There have been many findings that show the role of calcium binding proteins in abiotic stress tolerance like high salinity, drought and low temperature stress etc. Calcium binding proteins are one of the most important metallo-proteins known to play role during stress and also inhibits activity of other metallo-enzymes by competing for their binding site. In the present investigation, calcium binding protein containing EF-hand domain of five different species, i.e., Arabidopsis thaliana, Oryza sativa, Zea mays, Glycine max and Populus trichocarpa were analyzed. Percentage of calcium binding proteins was found to be highest in Oryza sativa and least in Populus tricocarpa. Binding sites were higher in Zea mays as compared to others. An assay was done to analyze the competition of metal for binding sites using alkaline phosphatase. It was observed that Ca2+ does not competes with the K+ and Na+ for the binding sites and activity of alkaline phosphatase increases in the presence of calcium only. Percentage decrease in root length of Oryza sativa and Zea mays, was also analyzed. It was noted that percentage decrease was higher in Zea mays in comparison to Oryza sativa. Also correlation between the number of binding sites and the level of abiotic stress tolerance was analyzed.
PF303 (IPPC0380) Morphological, physiological and biochemical responses of maize plants towards nano-micronutrient fertilization of Fe, Cu and Zn Rajamanickam Elanchezhian1*, Dameshwar Kumar2, Kulasekaran Ramesh1, Ashish Biswas1 and Ashok Patra1 1Indian Institute of Soil Science, Nabi Bagh, Berasia Road, Bhopal-462038, Madhya Pradesh, India 2Indira Gandhi Agricultural University, Raipur-492012, Chhattisgarh, India *Presenting author: [email protected]
Controlled release of fertilizer inputs for enhanced plant growth and improved nutrient utilization can be achieved through advanced nanotechnological tools. The impact of nano-micronutrient fertilization on growth and metabolism of maize plants was studied using Fe, Cu and Zn nanoparticles (NPs) based formulations. It was observed that there were differential plant responses towards the graded concentrations of NPs viz. Fe, Cu and Zn. The responses were evident on morphological, physiological and biochemical characteristics of maize. The nano-micronutrient fertilization of plants with lower concentration of Fe NP (27μM) had positively influenced photosynthetic leaf area, leaf area index, leaf area ratio and proline content of plants. However, plants treated with higher concentration of Fe NP (54μM) had exhibited improved growth parameters viz. plant height, shoot and root biomass, besides enhancement in relative growth rate, crop growth rate, net assimilation rate and catalase enzyme activity. In contrast to above, plants treated with Cu NP (0.5 & 0.25μM) had enhanced root length, membrane stability, total soluble sugar, non-structural carbohydrates, chlorophyll content while Zn NP (2 & 1μM) had accelerated the biochemical metabolism of plants. The above findings indicated that the effect of NPs including its dose was crop specific. Moreover it is also observed that NPs at reduced concentration may be useful for the crop and they may act as catalyst for growth and metabolism which can be utilized for higher yield of plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 268 Session F : Plant Nutrition Physiology and Genetics
PF304 (IPPC0384) Effect of salt on physiological responses of groundnut to iron deficiency Anita Mann1*, P.V. Zala2, A.L. Singh2, Vidhya Chaudhari2 and C.B. Patel2 1Division of Crop Improvement, ICAR-Central Soil Salinity Research Institute, Karnal-132001, India 2ICAR-Directorate of Groundnut Research, Junagadh-362001, Gujarat, India *Presenting author: [email protected]
Iron chlorosis is very common on alkaline soils such as calcareous ones, since iron availability is limited by high pH. Under these conditions of iron deficiency, graminaceous plant species induce special mechanisms for iron acquisition, involving enhanced release of iron chelators. On the other hand, it is known that most of salt soils have alkaline pH. So, plants growing on this kind of soils are often subjected simultaneously to salinity and iron deficiency. Thus, this work aimed at studying the physiological responses of groundnut (Arachis hypogea L.) to iron deficiency with salinity and evaluating the effect of salt on the iron nutrition and mineral nutrients. For this purpose, seedlings of two groundnut genotypes, GG-20 and TG-37 were cultivated under controlled conditions, either in a complete nutrient solution with or without NaCl, with +Fe and –Fe nutrient solutions. The plant morphological aspects, chlorophyll content of young leaves, iron status, water potential and mineral nutrient contents were assessed. Plants subjected to Fe deficiency exhibited a severe chlorosis, accompanied by a significant biomass reduction. Iron-deficient treatments (-Fe and –Fe/NaCl treatments generated Fe-deficiency chlorosis on young leaves. Visual Fe deficiency symptoms were observed after one week of growth under Fe starvation. However, the latter was greatly diminished when iron deficiency was associated to salinity. A depressive effect of salt on iron acquisition in plants subjected only to salt stress was observed which was further confirmed by significant decreased efficiency in iron acquisition. These results suggest that salinity may reduce capacity of plants to acquire iron from alkaline soils.
PF305 (IPPC0396) Comparative analysis of root proteome in soybean genotypes with contrasting exudation potential under phosphorus starvation Krishnapriya V. and Renu Pandey* Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Root exudation of carbon compounds is one of the mechanisms evolved by crop plants to adapt to low phosphorus (P) availability. High-molecular weight secretory proteins released by plant roots are involved in hydrolysing P from the organic pool, while low-molecular weight exudates such as carboxylic acids, phenols, free amino acids and sugars mobilise P from inorganic sources. This study aims to understand the physiological and molecular basis of root exudation in response to low P stress in soybean. Two soybean genotypes exhibiting contrasting root exudation potential (EC-232019 and EC-113396) were identified from 116 diverse genotypes. Higher P acquisition efficiency observed in EC-232019 compared to EC-113396 was attributed to greater release of root exudates and improved root architecture. EC-232019 exuded more carboxylates, proteins, phenols, sugars and free amino acids at low (4 μM) compared to sufficient (250 μM) P. Root morphological traits viz., root length, surface and volume were higher in EC-2302019 under low P. Further, when grown in soil with low (2 g P kg-1 soil) P, EC-232019 exhibited least reduction in leaf area, photosynthetic rate, total biomass, total P uptake and seed yield compared to sufficient (15 g P kg-1 soil) P. Comparative analyses of root proteome revealed over 10% of proteins detected on two-dimensional electrophoresis gels with two-fold or more altered expression between the genotypes. Out of approximately 225 protein spots detected, 28 differentially expressed proteins represented a large range of functional categories including carbon and energy metabolism, signal transduction and cell cycle regulation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 269 Session F : Plant Nutrition Physiology and Genetics
PF306 (IPPC0397) Regulation of iron deficiency tolerance by sulphur in wheat Vasundhara Sharma1* and Bhupinder Singh2 1Division of Plant Physiology, 2Nuclear Research Laboratory, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Deficiency of micronutrients in soil particularly, that of iron (Fe) is a major nutritional and production constraint worldwide, as these play significant role in plant metabolism. Present study elucidates the role and physiological basis of sulphur (S) mediated regulation of Fe uptake and use efficiency under deficient and sufficient availability of S (0, 1.2 and 2.5 mM) and Fe (1 and 100 μM) using nutrient solution culture in bread (HD-2967) and durum (HI-8713) wheat, which differ distinctly in their Fe deficiency tolerance response. Bread wheat, than durum wheat, exhibited a better plant growth under Fe deficiency which was related to the former’s ability to tolerate Fe deficiency through the production and release of a higher amount of phytosiderophores (PS) in the rhizosphere. Diurnal variation for the PS release under different treatments of S and Fe was similar in both wheat types. Further, the transcript expression of S transporter (SULTR1;1) and Fe transporter (YS1) revealed that SULTR1;1 transient expression requires not only low level of S but also sufficient Fe while the reverse was true for YS1 expression. Results clearly indicate a complex interplay of physiological, transcriptional and translational factors that governs the interaction between Fe and S metabolism and subsequently determine the regulatory role of S nutrition on Fe deficiency tolerance. Further, the requirement of sufficient Fe for the induction of high affinity S uptake transporter SULTR1;1 is important to gain insight into the regulation of S uptake by Fe.
PF307 (IPPC0414) Physiological and quality response of bartlett pear to calcium chloride sprays prior to harvest F.A. Khan*, Hafiza Ahsan, M.A. Dar and S. Narayan Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, Srinagar-190025, (J&K), India *Presenting author: [email protected]
Influence of foliar application of CaCl2 few days before harvest on post-harvest quality of Bartlett pear was investigated.
Plants were sprayed with CaCl2 0.0, 1.0 1.5 and 2.0% at 3, 2 and 1 weeks before harvest. The fruits were harvested at 110 days after full bloom. Bruise and disease free fruits were packed in corrugated fiber board boxes and stored at 1-2°C with 85–90% RH. Fruits showed continuous increase in PLW, TSS, sugars and spoilage with decreased firmness, ascorbic acid, phenol and acidity during storage and shelf life (20°C). Sprays of CaCl2 close to harvest proved advantageous in terms of increased fruit firmness, ascorbic acid, phenols and acidity with decrease in PLW, TSS and spoilage. The beneficial effects of CaCl2 also included reduction in respiration and ethylene production rates and retardation of skin yellowing. Spray of 1.5% CaCl2 before 14 days of harvest resulted in minimum PLW (9.12%), TSS (15.6%) and total -1 -1 -1 -1 sugars (12.1%), and respiration (7.3 ml CO2 kg hr ) and ethylene evolution rates (25.6 μl kg hr ) with maximum fruit firmness (11.6 lb inch-2), at 30 days of storage. Calcium treatments (1.5%) also caused an enhanced useful life of Bartlett pear by about 5-6 days as compared to control.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 270 Session F : Plant Nutrition Physiology and Genetics
PF308 (IPPC0432) Effect of putrescine (Put) supplementation and Arbuscular mycorrhizal (AM) fungi on growth, nutrient status and nodule symbiosis in Cajanus cajan (L.) Millsp (pigeonpea) under salinity stress Neera Garg and Amrita Sharma* Department of Botany, Panjab University, Chandigarh-160014, India *Presenting author: [email protected]
Salt stress is an important abiotic factor limiting plant growth and productivity in arid and semi-arid regions. Put, a member of polyamines plays an essential role in imparting salt tolerance in crop plants. In addition AM have the potential to reduce negative effects of salinity and alleviate ionic imbalance. Few reports suggest that Put enhance mycorrhizal root colonization. However, roles of Put in plant-fungal symbiosis are not well understood. Aim of this work was to verify the effects of pre- sowing seed treatment with putrescine and AM (Rhizophagus irregularis) on plant growth, mycorrhizal dependence, nutrient balance and nodulation potential in two genotypes of pigeonpea (Pusa 2001- salt tolerant and Pusa 991- salt sensitive) under NaCl salinity (0-100 mM). Salinity had significant negative effects on roots than shoots which disturbed root to shoot ratio (RSR). The impact of salinity was directly proportional to increased Na ion uptake and reduced AM percent root colonization. The decline in root growth affected the nodulation ability of Sinorhizobium fredii AR-4 in both genotypes, more in Pusa 991 than Pusa 2001. Individual application of Putrescine and AM improved plant growth, nutrient status and significantly reduced Na+ uptake. AM was found to be more efficient in improving nodulation ability of Pusa 2001 than Put. In addition Put was highly efficient in improving mycorrhizal colonization and imparted significant salt tolerance to pigeonpea genotypes under combined treatments (Put + AM). Our results demonstrated that the benefits of AM inoculation under salt stress can be significantly enhanced through Put treatment.
PF309 (IPPC0473) Effect of biodeg products on growth and biochemical attributes of sugarcane Smita Singh1*, Anshu Singh1, Abhishek Sharma2, M.K Srivastava2, R.K Singh1, Asha Gaur1, S.K. Shukla1, S.P. Singh1, A. Chandra1 and Radha Jain1 1Indian Institute of Sugarcane Research, Lucknow-226002, India 2Biodeg Chemicals & Allied Industry & Kemtech Polymers, New Delhi, India *Presenting author: [email protected]
In sub-tropical India, cane and sugar productivity are undesirably limited by factors like low germination (35-40%) consuming more time (about 45 days), low tillering (main cane / tiller ratio1:2) leading to lower NMC, finally culminating in cane and sugar yield of about 55-60 t/ha and 6-7 t/ha, respectively. In present investigation, a field experiment was conducted at IISR farm to study the effect of Biodeg products on growth and biochemical attributes of sugarcane. These two products are enzyme based organic fertilizers obtained by Biodeg Chemical & Allied Industry & Kemtech Polymers, New Delhi. Treatments comprises of T1 (Biodeg 310, @ 4 kg/acre) and T2 (Bioreg 311, @ 3 kg/acre) with 50% NPK, T3 (Biodeg 310, @ 4 kg/acre) and T4 ( Bioreg 311, @ 3 kg/acre) with 100% NPK, T5 (Biodeg + Bioreg, @ 4+3 kg/acre) 50% NPK, T6 (Biodeg + Bioreg, @ 4+3 kg/acre), with 100% NPK and T7 and T8 (control) with 50% and 100% NPK respectively. Results obtained indicated improvement in bud sprouting, shoot population, leaf number, nitrate reductase (NR) activity, chlorophyll, carotenoids, SPAD reading, amino acids and IAA contents by application of both the products. T1 treatment showed highest bud sprouting (29.44%) and shoot population (333/plot), however, fresh weight and leaf number were maximum in T5 at grand growth stage. NR activity ranged from 4.92 to 18.22 ìg nitrite formed per 100 mg fwt, T4 showed highest activity. IAA and amino acids contents of shoot tip were highest in T4 (75.54 ìg IAA/100 mg fwt) and T6 (168 ìg aa/100 mg fwt) treatments, respectively.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 271 Session F : Plant Nutrition Physiology and Genetics
PF310 (IPPC0488) Assessment of genetic variation for nodulation potential in chickpea genotypes under field conditions Karivaradharajan Swarnalakshmi1*, Subramaniam Gopalakrishnan2, Mohan Singh3, Sushil Kumar Chaturvedi3, Preeti Chopra1, Vijay Poonia1, Hari Deo Upadhyaya2 and Partha Basu3 1Indian Agricultural Research Institute, New Delhi-110012, India, 2International Crops Research Institute for the Semi-Arid-Tropics, Patancheru-5020324, Andhra Pradesh 3Indian Institute of Pulses Research, Kanpur-208024, India *Presenting author: [email protected]
Chickpea is an important rainfed crop covering about 9-9.5 m ha area and is largely grown on sandy loam to heavy clay soil with seldom use of irrigation, fertilizers and agrochemicals. The crop has the inherent potential to assimilate atmospheric nitrogen due to symbiotic association with Mesorhizobium ciceri. The contribution of biological nitrogen fixation in chickpea crop is reported to be around 40-80 kg/ha depending on the plant genotype and soil type. Lower grain yields at field conditions have also observed due to poor nodulation and nitrogen fixation. Identification of high nodulating genotypes for breeding cultivars with greater nitrogen fixing capacity will be useful for sustenance of chickpea productivity under field conditions. Therefore, evaluating the extent of genotypic variation in nodulation potential and plant growth in available chickpea minicore collection is required for selection of parental line for high BNF. Efforts were made to phenotype chickpea minicore lines to identify genetic source for high nodulation potential under field conditions. Nodulation, growth and yield at different physiological stages were recorded. Large variation in nodule weight ranged from 2 to 67.5 mg/plant was observed at early vegetative stage. Chickpea lines ICC13816 (67.5 mg/plant) and ICC4973 (60 mg/plant) showed highest nodulation compared to the IG7087 and ICC2737 (2 mg/plant). Similarly shoot weight of these lines were 840 (ICC13816) and 505 mg/plant (ICC 4973) and 150 mg/plant (IG7087) and 365 mg/plant (ICC2737). Plant genotype dependent variations in nodulation potential of chickpea mini core accessions provide a scope for selecting efficient nodulating lines and using them in varietal improvement programme.
PF311 (IPPC0519) An efficient element-contamination-free hydroponic system for dissecting phosphate deficiency-mediated response of the root system and molecular traits in rice Manisha Negi1*, Raghavendrarao Sanagala1, Vandna Rai1, Shivendra Sahi2 and Ajay Jain1 1National Research Centre on Plant Biotechnology, IARI, New Delhi-110012, India 2Western Kentucky University, 1906 College Heights Blvd. Bowling Green, KY 42101, USA *Presenting author: [email protected]
Phosphorus (P) is pivotal for plant growth and development, and absorbed by root system in the form of inorganic phosphate (Pi). However, due to slow diffusion rates in rhizospheres it is often a limiting nutrient leading to loss of crop productivity. Root system often attracted the attention because it exhibits extensive plasticity to various environmental cues including Pi and other nutrient deficiencies. However, unlike Arabidopsis, rice is largely been grown in pot culture and /or hydroponics but growth conditions currently being employed are often prone to elemental contamination, and not to facilitate easy documentation of the different traits of roots. Here in this study an element-contamination free aerated hydroponic system was developed for detailed temporal (2 d, 4 d and 7 d) analysis of the effects of Pi deficiency on different traits of primary, seminal and adventitious roots, of rice seedlings. After 2 d treatment, no significant effects were observed on any of these root traits. However 4 d and 7 d treatments, exerted differential effect on these root traits. Further, ICP-MS analysis revealed noticeable influence on ionomic profiles of macro-(P, K, S and Mg) and micro-(Fe, Zn, Mn and Cu) elements during growth under P+ and P- conditions. Further, Pi deficiency trigger significant increase on several genes that play a key role in Pi acquisition and or sensing and signaling cascade. Overall, the study provided the empirical evidence towards the fidelity of the system for detail analysis of Pi deficiency on morphophysiological and molecular responses of rice.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 272 Session F : Plant Nutrition Physiology and Genetics
PF312 (IPPC0592) Integrated approach for maximization of seed yield in mustard [Brassica juncea (L.) Czern & Coss] R.S. Sain*, S.S. Bareth, H.S. Cheema and N.K. Gupta Seed Technology Research, Rajasthan Agricultural Research Institute (Sri Karan Narendra Agriculture University, Jobner) Durgapura, Jaipur-302018, Rajasthan, India *Presenting author: [email protected]
Quality seed is important for realizing the potential of all other factors without which the improvement on account of fertilizer, irrigation and pesticides will not pay the desired dividends to the farmers. To study the effect of integrated yield maximization practices in mustard crop an experiment was conducted at RARI, Durgapura, Jaipur from rabi 2012-13 to 2014-15. The experiment was laid down in a split plot design in four replications with two main treatments based on date of sowing and sub treatments based on nutrient management through soil application viz., recommended dose (RD) of
NPK; RDNPK + gypsum (RD); RDNPK + gypsum (RD) + Zn (ZnSO4 @ 25 kg/ha); RDNPK + gypsum (RD) Zn (ZnSO4 @
25 kg/ha + Fe (FeSO4 @ 25 kg/ha. Results showed that seed yield and per cent germination were affected only by nutrient management and days to maturity by date of sowing significantly. All other characters viz., days to flowering, plant height, 1000-seed weight, seedling length and seed vigour index were affected by both date of sowing (main effect) and nutrient management (sub effect) significantly. Seed yield was maximum (23.68 q/ha) in treatment RDNPK + Gypsum
+ Zn (ZnSO4 @ 25 kg/ha) + Fe (FeSO4 @ 25 kg/ha) being at par with treatment RDNPK + Gypsum + Zn (ZnSO4 @ 25 kg/ ha)). Thus, timely sowing and optimized nutrient management including critical micronutrients serves to enhances seed yield and improves quality parameters such as 1000- seed weight, germination per-cent, seedling length and seed vigour index.
PF313 (IPPC0600) Effect of abiotic stresses on expression of genes involved in nitrogen uptake and assimilation pathways in Brassica juncea L. Parul Goel* and A.K. Singh Department of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061, India *Presenting author: [email protected]
Nitrogen (N) is primary plant nutrient that plays a crucial role in determining plant growth and productivity. Abiotic stresses like salinity, drought and extreme temperature affects the process of nitrogen uptake and assimilation in plants. However, little is known about the regulation of N pathway genes at transcriptional level under abiotic stress conditions in Brassica juncea. We have identified and cloned genes encoding nitrate transporters (NRT), ammonium transporters (AMT), nitrate reductase (NIA), nitrite reductase (NIR), glutamine synthetase (GLN), glutamate dehydrogenase (GDH), asparagines synthetase (ASN) and glutamate synthase (GLU) from Brassica juncea L. var. Varuna. Expression of these genes was studied under Salinity (150 mM NaCl), Cold (4°C), Heat (42°C), and Osmotic (250 mM Mannitol) stress after 1hr and 24hr of treatments. Most of the genes encoding N transporters and enzymes of assimilation pathway were found to be downregulated under abiotic stress treatments. The expression of genes encoding BjNRT1.1 and BjNRT2.1, main members of low and high affinity nitrate transport systems, respectively and genes encoding enzymes of nitrate assimilation are severely downregulated after 24h of stress treatments. The expression of BjAMT1.1, BjGDH1.1 and BjASN2 was downregulated at both, 1hr and 24hr of all the stresses treatments. While, expression of BjGS1.1 and BjGDH1.2 genes was downregulated after 1hr and expression of BjNRT2.1, BjNR2, and BjNIR1, genes was downregulated after 24h of all the stress treatments. However, expression of BjNRT1.1, BjNRT1.5 and BjGDH2 was upregulated after 1h of all stress treatments, while no gene was found to be upregulated after 24h of stress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 273 Session F : Plant Nutrition Physiology and Genetics
PF314 (IPPC0645)
Effect of high (CO2) on membrane lipid remodeling and phosphorus uptake efficiency in wheat under phosphorus stress with different sulphur levels Milan Kumar Lal*, Krishnapriya Vengavasi, Lakshmi Raj, Sangeeta Khetrapal and Renu Pandey Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Phosphorus (P), a non-renewable resource is deficient in most of the agricultural systems while continued rise in atmospheric
(CO2) helps the plants, particularly C3 in more efficient acquisition and utilization of P. We studied the effect of high (CO2) and sulphur (S) levels on alteration of membrane lipid composition under P stress. Different levels of P (5 μM, low-P and
500 μM, sufficient-P), (CO2) (380 ppm, aCO2 and 700 ppm eCO2) and S (10 μM, low-S and 2000 μM, sufficient-S) were used on bread wheat (Triticum aestivum cv. PBW-396) and durum (T. durum cv. PDW-233). The total P uptake increased in plants grown under e(CO2) with sufficient-P and S (control) by 32% while in low-S it was 46% higher compared to a(CO2). In membranes, the amount of phosphalidylglycerol (PG) decreased while there were 2- to 2.5-fold increase in sulfoquinovosyldiacylglycerol (SQDG) due to interactive effects of low-P and e(CO2). In plants grown at low-S, the level of SQDG decreased in both varieties but the interactive effects of e(CO2) and low-S resulted in two-fold increase in PG. In response to nutrient deprived condition, transcript levels of TaNPC4 increased in both root and shoot, increase being 2.6- folds in low-P, 1.5-fold in low-P/S and only 1.3-fold in low-S. The transcripts of TaPLD4 were highly induced in both root and shoot tissues in response to e(CO2). Our finding in turn suggests that under high atmospheric (CO2) plants will respond to P deficiency by altering its membrane lipid composition provided there is sufficient S fertilization.
PF315 (IPPC0672) Genome-wide expression of transcriptomes under iron and zinc stresses revealed candidate genes for kernel iron and zinc accumulation in maize (Zea mays L.) Mallana Gowdra Mallikarjuna1*, Nepolean Thirunavukkarasu1, Rinku Sharma1, Kaliyugam Shiriga1, Shikha Mittal1, Firoz Hossain1 and Hari Shanker Gupta2 1Maize Research Lab, Division of Genetics, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India 2Borlaug Institute for South Asia, CG Block, NASC Complex, New Delhi-110012, India *Presenting author: [email protected]
Deficiency of iron (Fe) and zinc (Zn) causes micronutrient-malnutrition or hidden-hunger affecting millions of people globally including sub-Saharan Africa, South Asia and Latin America where, maize is one of the major sources of food and feed. Among several available approaches, genetic-biofortification is a sustainable and cost-effective approach in addressing hidden-hunger. Identification of key genes involved in Fe and Zn homeostasis is an important prerequisite to breed micronutrient-rich hybrids. A transcriptome assay was performed, with ~18,000 transcripts distributed across the maize genome, in a SKV-616 inbred after exposing the seedlings to three Fe and Zn stress treatments (+Fe –Zn, –Fe +Zn, –Fe –Zn) along with control (+Fe+Zn). A total of 306 and 715 genes expressed in root and shoot, respectively were found common across stress treatments, suggesting presence of common transporters controlling both minerals in root and shoot. Differential expression of genes under stress treatments revealed that Zn association in many pathways was more pronounced over Fe pathways and the physiological demand for these minerals in shoot was much higher than in root. Significant expression of genes involved in mugineic acid pathway, metal-transporters, and phyto-hormone metabolisms revealed the role of those pathways in understanding the physiological and molecular mechanisms of Fe and Zn homeostasis. Co-mapping of DEGs with known QTLs for kernel Fe and Zn concentration identified several candidate genes such as OPT, metal-binding-protein, nramp3 and NAS. The candidate genes identified in the present investigation could be used for breeding kernel Fe- and Zn-rich maize hybrids.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 274 Session F : Plant Nutrition Physiology and Genetics
PF316 (IPPC0799) Effect of Treated distillery spentwash on fodder maize growth and yield M.B. Doddamani1*, Edna Antony2 and S.K. Gali3 1Department of Environmental Science, College of Agriculture, University of Agricultural Sciences, Dhrwad-580005, Karnataka, India 2Indian Grassland and Fodder Institute, Regional Research Institute, Dharwad-580005, Karnataka, India 3Dean (Student Welfare), University of Agricultural Sciences, Dhrwad-580005, Karnataka, India *Presenting author: [email protected]
A field experiment on the utility of treated spentwash as source of plant nutrients on fodder maize (African Tall) was conducted in the year 2014 at MARS, University of Agricultural Sciences, Dharwad. The soil of the experimental field was Vertisol with clay texture, alkaline in reaction and low electrical conductivity. Graded doses of treated spentwash was applied alone and incombintaion with fertilizer, based on the nitrogen requirement of the crop. Forty per cent of the calculated required nitrogen through spentwash was applied 15 days before sowing and remaining 60% was applied after sowing. Treated spentwash treatment @ 1½ N through spentwash recorded higher plant height, stem diameter, number of leaves, leaf area, leaf area index, Number of days to 50% tasseling, dry forage yield, seed yield, crude protein and crude fiber and was on par with 1½ N through fertilizer treatment. The highest plant height (289.6 cm) was recorded with 1½ N through spent-wash followed by 1½ N through fertilizer (286.7 cm). Seed yield (12.5 q/ha), dry fodder (17.0 t/ha), crude protein and crude fibre content were also higher with 1½ N through spent-wash treatments. Similar trend was also noticed with respect to stem thickness and leaf to stem ratio and maximum numbers of leaves (13.8) as compared to local farmers practice. This study is centered on the concept that treated distillery spent wash could be used for one time soil application as a nutrient source without adversely affecting soil and crop health.
PF317 (IPPC0805) Cloning and characterization of salt and nutrient stress responsive TaCBL4 gene from wheat Lekshmy S.*, S.K. Jha2, C. Viswanathan1 and R.K. Sairam1 1Division of Plant Physiology, 2Division of Genetics, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Worldwide wheat production is impacted by various abiotic stresses, like water deficit, salt, high temperature and nutrient deficiencies. Genetic improvement of abiotic stress tolerance of wheat can be greatly aided by knowledge of abiotic stress signaling. Calcium is a ubiquitous signaling molecule mediating signal transduction pathways of diverse environmental stresses. Calcineurin B-like proteins (CBLs) are related to neuronal calcium sensors of animals. CBL- interacting protein kinases (CIPKs) act as targets of CBL proteins and mediate stress response pathways in many plants. However, little is known about CIPKs in wheat and very few CBL-CIPK components are characterized in wheat. CBL4 (Salt Overly Sensitive 3) gene is involved in modulating salinity tolerance of Arabidopsis and Rice. TaCBL4 gene homologous to salt stress responsive OsCBL4 was cloned from wheat and sequence characterized. Expression of TaCBL4 gene was up regulated by salt stress, nitrogen, phosphorus and potassium deficiency albeit the expression was higher under salt stress. It is possible that TaCBL4 might mediate crosstalk among different signaling pathways. This work may facilitate functional studies of TaCBL4 gene and is beneficial for further molecular breeding in wheat.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 275 Session F : Plant Nutrition Physiology and Genetics
PF318 (IPPC0849) Potassium induced changes in antioxidant system and growth performance of Trigonella foenum-graecum L. cultivar RMT Madhup Shrivastava*, Mohammad A. Ahanger, Megha Tittal, Surendra Argal and R.M. Agarwal School of Studies in Botany, Jiwaji University, Gwalior-474011, India *Presenting author: [email protected]
Potassium is important for plants because of its active involvement in many functions such as enzyme activation, osmotic adjustment, maintaining transmembrane gradient and uptake of deleterious ions like Na. Pot experiments were conducted to find out the effectivity of different potassium salts on certain components of nitrogen metabolism, antioxidant system and ultimately on growth and yield of Trigonella foenum-graecum L. Varying doses (20, 40, 60 kg ha-1) of three potassium salts i.e., K2O, KCl and KNO3 were applied and analysis was done at two different developmental stages i.e., vegetative (25 DAS) and pre flowering (40 DAS). Obvious increase in growth and chlorophyll content was recorded. Supplementation of potassium salts enhanced the activity of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in Trigonella foenum-graecum L. Added potassium enhanced contents of total phenols and tannins, thereby probably strengthening both the enzymatic as well as non enzymatic antioxidant system. Potassium application resulted in considerable increase in synthesis/accumulation of osmolytes like free amino acids, proline and free sugars which assumes special significance in growth under water stress conditions. Enhancement in the activities of nitrate reductase and aminotransferases was also noticed indicating better nitrogen assimilation in potassium supplied plants thereby ensuring better plant growth. Hence, the result indicate that potassium supplementation may contribute towards qualitative changes in Trigonella foenum-graecum L beneficial from pharmacognostic/ medicinal viewpoint.
PF319 (IPPC0876) Yield and water production functions of wheat (Triticuma eastivum L.) cultivars and response to exogenous application of thiourea and ortho-silicic acid Ratna Kumar Pasala*, P.P. Deokate, G.C. Wakchaure and R.L. Choudhary National Institute of Abiotic Stress Management (NIAM), ICAR, Malegaon, Baramati-413115, Pune, Maharashtra, India *Presenting author: [email protected]
Supplemental irrigation, drought tolerant cultivars and use of plant bioregulators are now being proposed as the key strategy to unlock the yield potential and stabilize the yield of wheat grown in rainfed areas. Experiments were therefore conducted during 2013-2015 using line source sprinklers (LSS) to determine the interactive effects of quantities of supplemental irrigation and exogenous foliar sprays of plant bioregulators- 10 mM thiourea (TU) and 32 ppm of ortho- silicic acid (OSA) on wheat (Triticuma estivum L.) varieties viz., HD 2189, LOK 1, NIAW 301, NIAW 34 and PBW 550. The irrigation quantities were: fully irrigated (I1: 31.8 cm); mild (I2: 28.9 cm); medium (I3: 25.9 cm and I4: 22.7 cm); and severe
(I5: 19.9 cm and I6: 17.2 cm) water stress conditions. The two year field experiments demonstrated that wheat cultivars varied in their response to water deficits and those e.g. NIAW-301 with higher water productivity under limited irrigation. Response to TU, OSA also varied across water regimes and was higher under moderate to severe stress. Foliar application of TU and OSA at root crown index, flag leaf and seed filling stages improved grain yield by 9 and 6 % at fully irrigated; 19 and 18% at medium stress; 17 and 12% at severe water stress conditions and water productivity 0.12- 0.10 mg h-1 with TU, 0.13-0.09 mg h-1 with OSA at fully irrigated and medium stress; 0.11-0.03 mg h-1 with TU, 0.12-0.03 mg h-1 with OSA at severe stress conditions. Our major finding was that TU and OSA has various degrees of potential efficacy in alleviation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 276 Session F : Plant Nutrition Physiology and Genetics
PF320 (IPPC0877) Physiological approaches for yield improvement in pigeonpea in Karanataka Mahalingappa Dhanoji* and Mukesh Kumar Meena University of Agricultural Sciences, Raichur-584104, Karnataka, India *Presenting author: [email protected]
A field experiment was conducted during kharif-2014-15 to know the effect of hardening and foliar nutrition of essential elements and PGR’s with a view to mitigate terminal drought and realise yield potential in pigeonpea. Research results data revealed significant (6.4%) increase in the seed yield in pigeonpea with CaCl2 @ 2.0% seed hardening over control. Further there was also statistical improvement in the seed yield due to foliar nutrition of (all 19 nutrients including micronutrients) as compare to control. On the other hand, among the PGRs spray, NAA @ 20 ppm recorded significant improvement in the seed yield followed by nitrobenzene, BA-6 and CCC over control and tricontinol and water spray. The Interaction effect due to hardening, foliar spray of essential nutrients and PGRs, revealed significant improvement in the yield with CaCl2 @ 2.0% seed hardening + foliar spray of nutrients (all 19 nutrients including micronutrients) + foliar spray NAA @ 20 ppm over control and water spray. The improvement in the seed yield due to hardening, foliar spray of essential elements and PGRs might be attributed to improvement in the mean value of ancillary growth and physiological parameters, viz., plant height, number of branches, test weight, pod weight per plant, Dry matter accumulation, light interception, NDVI and SPAD values.
PF321 (IPPC1008) Impact of elevated atmospheric carbon dioxide level on uptake of iron and zinc in rice crop A. Raj1, B Chakrabarti2, H. Pathak3, U. Mina4, R.C. Harit5* and V. Kumar6 Centre for Environment Science and Climate Resilient Agriculture, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Climate change associated with rise in atmospheric greenhouse gases (GHGs) concentration is likely to affect global food production and agricultural sustainability. Rice crop is also affected by this rise in carbon dioxide (CO2) in terms of growth, yield and nutrient uptake. An experiment was conducted during the kharif season of year 2013 inside and outside the Free Air Carbon dioxide Enrichment (FACE) facility at the farm of Indian Agricultural Research Institute, New Delhi to study the impact of elevated CO2 on uptake of iron (Fe) and zinc (Zn) in rice crop. Two N doses were applied in both -1 ambient (395 ppm) and elevated CO2 (550 ± 20 ppm) conditions i.e., N0: no N (control), N1: 0.8 g N pot (100 % recommended dose). Results showed that high CO2 significantly enhanced both grain and biomass yields of rice crop.
Uptake of Fe and Zn increased significantly with rise in CO2 level and with N application. With recommended dose of N, uptake of Fe and Zn in rice grain increased by 64.2% and 19.2% under elevated CO2 condition as compared to ambient
CO2 level. Total uptake of Fe and Zn also followed the same trend with 31.9% and 25.9% increase in elevated CO2 treatment. Increased yield at higher CO2 level has resulted in higher micronutrient uptake of rice crop.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 277 Session F : Plant Nutrition Physiology and Genetics
PF322 (IPPC1009) Ammonia volatilization under different air temperature regimes in wheat crop Renu Singh1*, Vijay Kumar1, Arti Bhatia1, Renu Pandey2, Vinod Kumar1, Monika Srivastava1 and Himanshu Pathak1 1Centre for Environmental Science and Climate Resilient Agriculture, 2Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Nitrogen (N) is an essential nutrient for crop production and its efficient use is important for the economic sustainability of cropping systems. Nitrogen use efficiency (NUE) is one of the major issues in agriculture as it is directly associated with crop yield, fertilizer costs and N-pollution. In the present scenario of global warming, the increasing air temperature influences NUE and ultimately crop yield. Ammonia volatilization is the major process leading to loss of N from soil under increasing temperatures. Wheat being major cereal crop, farmers is bound to apply excessive nitrogenous fertilizer to increase crop productivity which leads to low NUE as well as lower net returns. An experiment was conducted to evaluate the effect of high atmospheric temperature on ammonia volatilization and yield under different N levels in wheat crop.
Four different levels of temperature (ambient temperature, temperature gradient tunnel (TGTc) control, TGT+2.5°C and TGT+3.5°C) and N (0, 90, 120 and 150 kg ha-1) were taken in combinations to grow wheat. The ammonia volatilization loss ranged from 4.2-20.4 kg ha-1 season-1 while the highest loss was observed in treatment with 150 kg ha-1 N and TGT+3.5°C. This treatment accounted for 13.6% loss of applied N. The study revealed that the rate of volatilization significantly increased with rise in air temperature which may lead to decrease in NUE in winter crops like wheat thus reducing productivity.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 278 Session G : Plant Growth Regulators and Applications
PG323 (IPPC0097) Utilization of fungicide for better seedling growth responses in mung bean [Vigna radiata (L.) Wilczek] Mukul Kumar* and Prakash Chandra Gupta Bihar Agricultural University, Sabour, Bhagalpur-813210, Bihar, India *Presenting author: [email protected]
Fungicides are commonly used for seed treatment to prevent seed borne diseases, and thus to establish a good field stand for high productivity. Non-systemic fungicide thiram (N,N-dimethylcarbamodithioate) and systemic fungicide bavistin (methyl-N-1H-benzimidazol-2-yl-carbamate) are widely used for seed treatment. However, these fungicides may influence the plant physiological process and thus may regulate seedling vigour and crop growth. Previous study on wheat showed that seeds treated with these fungicide delayed germination and decreased percentage germination. In contrast, seed dressing with thiram has been shown to increase the seedling vigor of sorghum. Hence, we conducted a lab experiment to study the effect of thiram and bavistin seed treatment on the seedling growth of mung bean, a major rice fallow pulse in India. A lab experiment in CRD design with three replications was conducted under controlled temperature and relative humidity in lab conditions during kharif season. The experiment consisted of three treatments, viz., non-treated seeds (control), thiram and Bavistin treated seeds of mung bean. Morphological characters such as shoot length, root length, leaf area and dry weight of seedling showed significant differences between treatments. Thiram fungicide treatment- induced enhancement of the shoot length, root length, leaf area and dry weight of seedlings was higher than that of Bavistin treatment and control. The results suggest that species specific response of plants to these fungicides. Treatment with thiram may help enhance yield of rice-fallow mung bean in India.
PG324 (IPPC0111) Role of ethylene in heavy metal tolerance: Modulation of ascorbate-glutathione pathway and proline metabolism in mustard plants M. Iqbal R. Khan* and Nafees A. Khan Department of Botany, Aligarh Muslim University, Aligarh-202002, U.P., India *Presenting author: [email protected]
Heavy metal contamination of agricultural soil is primarily related to the abrupt anthropogenic activities. Plants frequently exposed to high concentration of HMs shows adverse effects. In the present study we investigated the role of ethylene significance in nickel (Ni) and zinc (Zn) stress tolerance in mustard (Brassica juncea) plants. Application of Ni and Zn at the rate of 200 mg kg-1 soil induced oxidative stress along with increased ethylene production. Under Ni and Zn stress, photosynthetic attributes and growth characteristics of mustard plants was reduced. Application of ethephon significantly increased ascorbate-glutathione (AsA-GSH) pathway and proline metabolism and maintained cellular homeostasis and water relation of mustard plants. Up-regulation of AsA-GSH and proline metabolism resulted in protection of photosynthesis and growth of the plants under Ni and Zn stress. The result was substantiated by using ethylene action inhibitor, norbornadiene (NBD), which significantly inhibits both AsA-GSH cycle and proline metabolism resulted in less protection of photosynthesis and growth of plants. The results of the present study could be exploited in situation where plants are grown under higher level of Ni and Zn conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 279 Session G : Plant Growth Regulators and Applications
PG325 (IPPC0129) Potential uses of homobrassinollide plant growth regulator in increasing berry growth and yield of grape var. Thompson seedless Ajeet Singh Tomar*, Burhanuddin Bohra and Girish Dalal Godrej Agrovet Ltd., R&D Department, Vikhroli-East, Mumbai- 400079, India *Presenting author: [email protected]
Plant growth regulators impart important role towards higher yield of crops. Among different strategies, exogenous applications of plant growth regulators are involved in promoting plant growth and development under normal and stressful conditions. Plant growth regulator containing growth hormone homobrassinolide (HBR) enables crop plants to overcome inefficiency barriers by triggering vital physiological process ensuring higher yields of agricultural and horticultural crops. In this study, possible role of exogenously application of HBR on berry growth, bunch weight and quality parameters of table grapes were evaluated. The applications of HBR alone and in combination with Gibberellic acid (GA) in the form of two dips were given at 2-3 mm and 5-6 mm berry size stages. Clusters were harvested when those of the control vines reached 16-17 °Brix. Yield and quality parameters were analyzed. It was revealed that first dip with HBR (1 ppm) + GA ( 50 ppm) at 2-3 mm berry size stage followed by second dip with HBR (1 ppm) + GA ( 40 ppm) at 5-6 mm berry size stage provides significant increase in berry weight, berry size and bunch weight over control and GA treatment. The study revealed that the use of HBR in combination with GA has a great significance due to its synergistic effects in quality production berries of grape var. Thompson seedless which has huge potential in export markets.
PG326 (IPPC0144) Multiple Interactions between glucose and brassinosteroid signalling in regulating plant growth and development Aditi Gupta*, Manjul Singh and Ashverya Laxmi National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India *Presenting author: [email protected]
Brassinosteroids (BRs) and sugars/glucose (Glc) are fundamental to plants and regulate similar processes. There are few evidences for BR and Glc signaling interactions in plants but its molecular basis is not explored. We hypothesize that there may be an extensive overlap between BR- and Glc- signaling pathways. Whole genome transcriptome analysis was performed to find out the interdependence/overlap between BR and Glc signal transduction pathways in light grown as well as etiolated seedlings of Arabidopsis. Glc alone was able to regulate 55% and 71% of BR-regulated genes under light and dark growth conditions respectively. BR and Glc commonly regulate a large number of genes involved in growth and development. Physiologically; both Glc and BR could regulate many aspects of early seedling growth and development such as; hypocotyl and root elongation, root and shoot directional growth, lateral root emergence, root hair development etc. We utilized both genetic as well as pharmacological approaches to establish hierarchy of multiple signaling components such as Glc, BR, auxin, cytokinin ethylene etc. in regulating different parameters of seedling development. Briefly, BR signaling via BRI1 receptor interacts with HXK1-dependent Glc signaling pathway. Intact BR response pathway is essential for Glc regulation of seedling growth and development. Auxin transport and signaling components mainly act as nodal points in regulating Glc and BR mediated growth responses. Our results, altogether, suggest that Glc and BR signaling pathways interact extensively to regulate various growth parameters required for maintaining optimal seedling fitness in nature.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 280 Session G : Plant Growth Regulators and Applications
PG327 (IPPC0159) Protection of growth via antioxidants and endogenous hormones maintenance in response to 24- epibrassinolide in tomato (Lycopersicon esculentum L.) growing under salt, drought and heavy metal stress Deepak Singh Bagri* and Chandrama Prakash Upadhyay Department of Biotechnology, Dr H.S. Gour Central University, Sagar-470003, Madhya Pradesh, India *Presenting author: [email protected]
Brassinosteroids (BRs) are a group of plant growth steroidal hormones present in very low concentrations in seeds, pollen, young plant tissues and are reported to be involved in plant development as well as in protection of plants against various biotic and abiotic stresses. In this study, we report the effect of epibrassinolide (EBR) on in vitro and ex vitro grown tomato (Lycopersicon esculentum L. cv. Pusa Ruby) seedlings under induced salt (NaCl), drought (mannitol) and heavy metal (ZnCl2) stresses. These stresses hampered the plant development and plant showing decreased shoot length, total leaf area and plant dry matter. However, these parameters changed with application of EBR and enhanced values with the same parameters were observed under both in vitro and ex vitro conditions. Significant increases in total leaf chlorophyll, proline content and nitrate reductase (EC 1.6.6.1) activities were observed in leaves of tomato plants treated with EBR when compared to their respective stressed controls. The activities of antioxidant enzymes such as ascorbate peroxidase (EC 1.11.1.11), catalase (EC 1.11.1.6), dehydroascorbate reductase (EC 1.8.5.1), glutathione reductase (EC 1.6.4.2) and superoxide dismutase (EC 1. 15.1.1) enhanced in tomato plants under these stresses and their activities further enhanced with the application of EBR. The endogenous levels of indole-3-acetic acid and abscisic acid elevated significantly in the leaves of tomato plants treated with EBR compared to stressed controls. This is the first comprehensive report confirming the protective role of EBR in imparting tomato plant tolerance to abiotic stresses under both in vitro and ex vitro conditions.
PG328 (IPPC0186) Silencing of OsCKX2 enhances panicle branching resulting in improved grain yield in rice under both normal and salinity stress conditions Rohit Joshi1*, Khirod Kumar Sahoo1, Amit Kumar Tripathi1, Ashwani Pareek2 and Sneh Lata Singla-Pareek1 1Plant Molecular Biology, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India 2Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Plants maintain their cytokinin levels through cytokinin oxidases which catalyze irreversible degradation of cytokinins. Rice cytokinin oxidase 2 (OsCKX2) gene has earlier been identified as a major QTL playing crucial role in determining grain number. In the present study, we have carried out in-depth studies to unravel the physiological and developmental function of OsCKX2. Via expression analysis using qRT-PCR, we found that OsCKX2 expression is regulated in a developmental-stage specific manner. The highest level of OsCKX2 transcript was found between 10-13 days after panicle initiation. To further study the effect of reduced OsCKX2 levels, stable RNA interference mechanism was utilized for silencing OsCKX2 gene in rice. Ultra-performance liquid chromatography (UPLC)-based estimation of CKs revealed a significant increase in CKs in OsCKX2-knockdown plants. Enhanced CK levels correlated with enhanced grain yield in the OsCKX2-knockdown plants. To check if the OsCKX2-knockdown plants maintain their yield even under salinity stress conditions, we assessed their growth, physiology and grain yield under these conditions. OsCKX2-knockdown plants showed better relative water content, higher photosynthetic efficiency and reduced electrolyte leakage as compared to the wild-type plants under 200 mM NaCl stress. Importantly, we found a negative correlation between OsCKX2 expression and plant productivity as the OsCKX2-knockdown plants showed enhanced panicle branching, more filled grains per plant, higher 1000 grain weight and better harvest index. Together, the results of our study suggest that OsCKX2 may be explored as a unique regulator of floral primordial activity for enhancing grain production under normal as well as abiotic stress conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 281 Session G : Plant Growth Regulators and Applications
PG329 (IPPC0291) Auxin plays a critical role in spikelet fertility of rice under drought and high temperature stresses Laxmi Sharma1*, Rakesh Verma2, Santosh Vinjamuri2, S.R. Kushwaha2, Monika Dalal3 and Viswanathan Chinnusamy2 1ICAR-Vivekanand Parvatiya Krishi Anusandhan Sansthan, Almora-263601, Uttarakhand, India 2Division of Plant Physiology, 3NRC on Plant Biotechnology, ICAR-Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Reproductive processes of plants are highly vulnerable to the environmental stresses, which are increasing due to global climate change. Drought and high temperature (HT) are the key stress factors limiting the yield potential in rice. Auxin is considered to be the master regulator of development and patterning. Increasing evidences indicate that auxin plays critical role in adapting the abiotic stress response. Reports show that HT-induced reduction in the endogenous auxin content was responsible for loss in pollen viability in barley and Arabidopsis. In this study, we analysed the role of auxin (indole-3-acetic acid, IAA) in regulating the spikelet fertility of rice under drought and HT stresses. Both these stresses reduced endogenous IAA levels and the expression of genes coding for auxin biosynthesis and signalling. In addition, these stresses significantly reduced pollen viability and spikelet fertility in most of the rice genotypes. Application of exogenous IAA before exposing the plants to drought or HT stress mitigated adverse effect of drought and heat through restriction of oxidative stress. This was reflected in better pollen viability and spikelet fertility percentage as well as lower ROS accumulation and membrane injury in the stressed plants. These results suggest that stress induced decline in endogenous IAA levels is one of the causes of reduced spikelet fertility in rice. Exogenous application of auxin and/or genetic improvement in the ability of rice to maintain auxin homeostasis under these stresses could help maintain the spikelet fertility and thus combat the global warming-mediated yield loss in rice.
PG330 (IPPC0345) Physiological effects of strobilurin compounds in chilli (Capsicum annum L.) A. Senthil*, S. Punitha and P. Jeyakumar Department of Crop Physiology, Tamil Nadu Agricultural University, Coimbatore-641003, Tamil Nadu, India *Presenting author: [email protected]
Strobilurins, a new class of fungicides, have shown to be effective in altering the physiological and biochemical traits of crop plants. Experiment was conducted to study the influence of strobilurin fungicides on morpho-physiological, biochemical parameters and yield of chilli. The experiment consisted of seven treatments with strobilurin such as trifloxystrobin combined with tebuconazole as Nativo @ 300, 350 and 400 g/ha, pyraclostrobin combined with metiram as Cabriotop @ 1500 g/ha and azoxystrobin as Amistar @ 500 ml/ha, along with carbendazim @ 500 g/ha and untreated control. The treatments were given as foliar spray at 45 and 60 DAT of chilli crop. The results revealed that application of Nativo @ 400 g/ha recorded the highest root length, root shoot ratio and more number of branches per plant among all the treatments. Maximum leaf area, LAI and SLW were recorded in Cabriotop @ 1500 g/ha followed by Nativo @ 400 g/ha. The CGR and TDMA were significantly influenced by Nativo @ 400 g/ha. Among the treatments, Nativo @ 400 g/ha recorded the highest chlorophyll index, photosynthetic rate and recorded the maximum green chilli yield of 20.44 t/ha which was followed by Cabriotop @ 1500 g/ha (19.40 t/ha). Hence, it is concluded that foliar application of trifloxystrobin combined with tebuconazole as Nativo @ 400 g/ha at 45 and 60 DAT was found optimum for increased yield with high economic returns in chilli crop.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 282 Session G : Plant Growth Regulators and Applications
PG331 (IPPC0372) Effect of foliar application of plant growth regulators on photosynthetic rate, stomatal density and grain yield in relation to nitrous oxide (N2O) emission from field grown wheat Nirmali Bordoloi* and K.K. Baruah Department of Environmental Science, Tezpur University, Tezpur-784028, Assam, India *Presenting author: [email protected]
Agricultural fields are the major anthropogenic source of greenhouse gases nitrous oxide (N2O) and methane (CH4) emission to the atmosphere. The N2O emission from wheat grown field was studied for two years to investigate the impact of different plant growth regulators (PGR) on photosynthesis, grain yield and N2O emission reduction. The growth regulators namely Abscisic acid (ABA) and Cytozyme (20 ppm), Kinetin (10 ppm and 20 ppm) and wet tea extract (1:20 w/w) along with distilled water as control were sprayed on wheat canopy at tillering and panicle initiation stages and N2O emission was estimated from the treated plants. Cytozyme and tea extract markedly enhanced the plant dry weight in term of biomass over control, while abscisic acid and kinetin increased the plant photosynthetic rate at panicle development and flowering stages. The increased rate of photosynthesis in response to ABA and kinetin indicate efficient photosynthate partitioning towards the developing grain. ABA (20 ppm) and Kinetin (10 ppm) also decreased the N2O emission over control along with better grain production. Leaf area index, stomatal conductance, stomatal frequency and xylem vessel size are found to be associated with N2O transport and emission to the atmosphere. Use of selective PGR like ABA and
Kinetin can enhance the grain productivity of wheat and also reduce the N2O emission from wheat cropping system.
Approaches to increase wheat productivity and reduce N2O emission may be particularly beneficial in a future climate with rising temperature.
PG332 (IPPC0387) Seasonal effects of explant collection on bud break and culture establishment from mature nodes of Tecomella undulata (Sm.) Seem. Sidhika Chhajer, R. Sharma, R.K. Bhatt and Rajwant K. Kalia* Central Arid Zone Research Institute, Jodhpur-342003, Rajasthan, India *Presenting author: [email protected]
Tecomella undulata (Family Bignoniaceae), a medicinally and economically important timber tree of the dry regions of India, Pakistan and Arabia, has been considered as endangered due to over exploitation and negligible conservation efforts. Micropropagation can produce large number of disease free clones, therefore, to study the seasonal effect of explant collection on bud break and shoot length, nodal segments were collected from 31 year old trees every month and cultured on MS medium supplemented with BAP (10 uM), ascorbic acid (50 mg/l), arginine (25 mg/l) and citric acid (25 mg/l). Highest percentage of explants (73%) showing response was recorded in explants collected during the months of August-September. Fresh flush of shoots sprouted after rains in July are best for in vitro establishment of cultures. Decline in response from November to February could be due to transition of actively growing spring buds to winter buds during which cytokinin supply to shoot system falls down. Decreased response in the months of March-May could be due to sinking of nutrients in pods for filling. Explants collected during rainy season were more prone to contamination while it was not a major problem in other months. Shoots collected during dormant months showed increased callusing at the explant base with lesser bud induction. A progressive reduction in callusing at explant base and increase in number and length of sprouted buds was observed in explants collected during months of active growth. Therefore, the study concluded that explants collected from actively growing shoots were best for culture establishment.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 283 Session G : Plant Growth Regulators and Applications
PG333 (IPPC0390) Influence of different 1 methyl cyclopropene doses and storage conditions on quality and organoleptic evaluation of alphonso mangoes Nitin Gokhale1*, Arun Mane1, M.B. Chetti2, Sandip Mahadik1, Murad Burondkar1 and Sanjay Bhave1 1Dr. B.S. Konkan Krishi Vidyapeeth, Dapoli-415712, Maharashtra, India 2Indian Council of Agriculture Research, Krishi Anusandhan Bhawan-II, New Delhi-110012, India *Presenting author: [email protected]
Investigations were carried out, for two consecutive years (2011 and 2012), with an objective to test the effect of 1-methyl cyclopropene (1-MCP) under different storage conditions on physico-chemical properties and organoleptic evaluation of Alphonso mango fruit observed after prolonged shelf life during transportation. After harvesting at physiological maturity, followed by pre-cooling at 130C, fruit were exposed to vapors of 1-MCP @ 70 mg/m3 (1000 ppb) for 18 hrs in air tight (plastic) treatment chamber. The fruit quality assessed at different intervals viz. after 7, 14, 21 and 26 days of storage and shelf life behavior. Treatment combination of 1-MCP at 2000 ppb concentration and cold storage at 13 ±10C found effective in maintaining the quality of fruits even after 26 days storage (17.90 0B, 0.38%. 12.50%, 4.63% respectively) as compared to control in 21 DAT (12.62 0B, 0.22%, 12.95, 3.875% respectively). The fruits treated with higher concentrations of 1-MCP and kept under cold storage yielded slow rise in vitamin A content with further decline after reaching the peak values till the end of storage whereas the ascorbic acid content untreated ambient control fruits was significantly decreasing as compared to other treatment combinations. The organoleptic evaluation obtained at optimum ripening shown no significant difference in taste, aroma and flavor amongst all the treatment combinations, whereas 1-MCP treated fruits with cold storage shown less occurrence of spongy tissue (physiological disorder) with better colour visibility when fruits were treated with 2000 ppb 1-MCP.
PG334 (IPPC0391) Ripening and storage behavior of alphonso mangoes as influenced by 1 methyl cyclopropene doses under different storage conditions Arun Mane1*, M.B. Chetti2, Sanjay Bhave1, Murad Burondkar1 and Nitin Gokhale1 1Dr. B.S. Konkan Krishi Vidyapeeth, Dapoli-415712, Maharashtra, India 2Indian Council of Agriculture Research, Krishi Anusandhan Bhawan-II, New Delhi-110012, India *Presenting author: [email protected]
Investigations were carried out for two consecutive years (2011 and 2012), with an objective to test the efficacy of different doses of 1-methyl cyclopropene (1-MCP) under different storage conditions, in reducing the ripening rate and prolonging the storage and shelf life of Alphonso mango fruit to suit export by sea transportation from India. After harvesting at physiological maturity, followed by pre-cooling at 130C, fruit were exposed to vapors of 1-MCP @ 70 mg m-3 (1000 ppb) for 18 hrs in air tight (plastic) treatment chamber. The treated and untreated fruits were then transferred to different storage conditions for different intervals (7, 14, 21, and 26 days). Effect of 1-MCP on ripening pattern of fruits examined at six difference stages (viz. green, turning yellow, ½ ripe, ripe, diseased and shrivelled) indicated that 1-MCP treatment significantly arrested the ripening till 8 days and prolonged the subsequent stages from 12, 16, 20 and 24 days, over untreated control fruit. The physical changes evaluated at different intervals shown the reduction in loss of colour, fruit firmness and physiological weight under cold storage with an effective dose of 1-MCP at 2000 ppb, ultimately increasing the shelf life of Alphonso mangoes. Whereas, physiological changes such as chlorophyll content, total carotenoids, rate of respiration and release of ethylene recorded at different intervals viz., after 7, 14, 21 and 26 days of storage found to sustain the minimum values till the end of storage period in case of cold storage treatments with 1-MCP at 2000 ppb concentration.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 284 Session G : Plant Growth Regulators and Applications
PG335 (IPPC0401) Juglone promotes shooting and inhibits rooting in leaf explants of in vitro raised tomato (Lycopersicon esculentum Miller var. Pusa Ruby) seedlings Kiran Bamel1*, Rajendra Gupta2 and Shrish C. Gupta2 1Department of Botany, Shivaji College (University of Delhi), Raja Garden, Delhi-110007, India 2Department of Botany, University of Delhi, Delhi-110007, India *Presenting author: [email protected]
The effect of juglone was evaluated on morphogenesis of leaf explants excised from in vitro raised 30-d-old seedlings of tomato, Lycopersicon esculentum var. Pusa Ruby. To test the dose response different levels (10-9-10-3 M) of juglone was added to the controls. Juglone caused significant inhibition of rhizogenesis as reflected by the reduced rooting potential of the leaf explants and transition from rooting to shoot formation at 10-7-10-6 M. Juglone increased the caulogenesis and callusing when added to a shoot regeneration medium. The optimum response was observed at 10-5 M. The results of the present investigation indicated that juglone acts as a stimulator of callusing and shoot formation.
PG336 (IPPC0409) Effect of JA and Me-JA on chlorophyll fluorescence and photosynthetic pigments of three cultivars of Brassica oleracea L. Geetika Sirhindi* and Ruqia Mushtaq Department of Botany, Punjabi University, Patiala-147002, India *Presenting author: [email protected]
The present study focuses on the effect of Me-JA and JA on chlorophyll fluorescence and photosynthetic pigments in three cultivars of Brassica oleracea L. cv. Italica, Capitata and Botrytis. In cv. Botrytis maximum yield of PSII (Fv/Fm) was enhanced by 17%, 13% and 12% in 1μM Me-JA, 1 nM Me-JA and JA treated seedlings, respectively. Effective quantum yield [Φ PSII (Y)] enhanced in all treatments of Me-JA and JA in Botrytis. No effective enhancement of Me-JA and JA treatment was experiential in Italica and Capitata on Fv/Fm and Φ PSII (Y). JA and Me-JA augmented photochemical quenching (qP) in Italica to 281% in 1 nM JA. In Capitata 1μM JA and 1 nM Me-JA showed 14% and 21% enhancement in qP. Application of JA and MeJA decreased non photochemical quenching (NPQ) in Capitata and Botrytis, but in Italica 1 pM JA and MeJA increased NPQ by 111% and 67% respectively. Total chlorophyll, chl a, chl b and carotenoids showed significant enhancement in Italica in presence of JA and MeJA whereas in Capitata and Botrytis this enhancement was in 1 nM JA. From the above results it can be suggested that MeJA and JA have a potential to protect the plant from photoinhibition but in dose dependent manner and also depends on cultivar.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 285 Session G : Plant Growth Regulators and Applications
PG337 (IPPC0428) Isolation and characterization of phosphate solubilising bacteria from sugarcane rhizospheric soil for screening of plant growth promoting activities Deepmala Katiyar1*, A. Hemantaranjan1 and Bharti Singh2 1Department of Plant Physiology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi- 221005, Uttar Pradesh, India 2Department of Microbiology, Institute of Medical Sciences, Banaras Hindu University, Varanasi-221005, Uttar Pradesh, India *Presenting author: [email protected]
Now a days new practices using beneficial microbes are being developed and employed for sustainable agriculture. The present study is limited to plant growth promoting rhizobacteria (PGPR). PGPRs are a group of beneficial soil bacteria that can be found in the rhizosphere, in association with roots which can enhance the growth by a wide variety of mechanisms. The use of PGPR is gradually increasing in agriculture and offers an attractive way to replace chemical fertilizers. A large number of bacteria including species of Pseudomonas, Azospirillum, Azotobacter, Klebsiella, Enterobacter, Bacillus and Rhizobium have reported to enhance plant growth. The study was conducted to isolate phosphate solubilising bacteria (PSB) from sugarcane rhizosphere and investigate their potential for plant growth promotion. Twenty nine PSB were isolated by serial dilution in selective media from three places of Uttar Pradesh. Isolated rhizobacteria were characterized by morphological, physiological and biochemical method. The study was further preceded for molecular identification of bacteria by 16S rRNA sequencing, and in vitro screening of plant growth promoting traits viz., production of Indole acetic acid (IAA), Hydrogen cyanide (HCN) Ammonia (NH3) production and antifungal activity against Colletotrichum falcatum. Seven isolates (PSC1, PSC2, PSC3, KSC1, KSC2, P13, P35) containing Enterobacter sp., Pseudomonas sp. and Bacillus sp. have showed high ability in phosphate solubilising and plant growth promoting activities out of 19 isolates. Enterobacter sp. (PSC3) showed highest plant growth promoting activities among three rhizobacteria. This study, therefore, suggest that the use of PGPR isolates as inoculants might be beneficial for sugarcane cultivation.
PG338 (IPPC0434) Intrinsic salicylic acid mediated delay in postharvest life of tomato: ROS metabolism Tushita Shukla*, Chunoti Changwal, Ajay Arora and V.P. Singh Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Postharvest issues are common around the world. The extent of the loss of horticultural products after harvest can vary in different countries. A remarkable association between exogenous salicylic acid (SA) and fruit ripening has been reported in various fruits including tomato. SA is known to affect various physiological and biochemical activities of plants and may play a key role in regulating their growth and productivity. It behaves in a concentration and time dependant manner. Tomato is a good model system for studying the ripening process. The endogenous SA is directly correlated with postharvest life of tomato, this hypothesis, has to be tested in 55 tomato germplasm at different stages of fruit ripening vis-á-vis shelf life. We screened 15 contrasting tomato germplasm for further validation of the hypothesis in association with ROS metabolism which has been reported previously that external application of SA can alleviate the oxidative stress and enhance the antioxidative capacity. We observed that oxidative stress increased from green to red ripen stage of tomato and antioxidative defence rise up to certain level then it decline in later stages of tomato. We will discuss the potent ROS species, antioxidants (enzymatic or nonenzymatic) and osmoprotectants that play key role in process of ripening in comparison to SA and shelf life.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 286 Session G : Plant Growth Regulators and Applications
PG339 (IPPC0435) Effect of salicylic acid on physiological traits, antioxidant enzymes and grain yield of wheat genotypes under water deficit stress conditions D.C. Saxena1*, S.V. Sai Prasad2, Iti Rathi2 and Renu Parashar2 1Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India 2IARI-Regional Station, Indore-452001, M.P., India *Presenting author: [email protected]
Drought or water deficit stress is the main constraint limiting crop production globally. An experiment was conducted to study the effect of exogenous foliar application of salicylic acid (SA) (0.5 mM concentration) applied at anthesis stage of five contrasting wheat genotypes viz. HI 1581, C 306, Raj 4083, HI 8731 and NI 5439 under water deficit stress conditions. Physiological traits viz. relative water content, membrane stability index and chlorophyll content were decreased under water deficit conditions, but the effect was mitigated in salicylic acid treated plants and there was increase in these characteristics. The activities of antioxidant enzymes viz. superoxide dismutase, catalase, peroxidase and ascorbate peroxidase was upregulated by water deficit stress and further enhanced by SA treatment which in turn protected plants against ROS generation and membrane injury. The yield attributes also showed significant reduction under water deficit stress whereas application of SA enhanced the yield and its components. Hence, salicylic acid can ameliorate the deleterious effect of water deficit stress by increasing the physiological characteristics and antioxidant enzymes.
PG340 (IPPC0448) Endogenous salicylic acid associated regulation of fruit ripening in tomato: ethylene and softening enzymes Chunoti Changwal*, Tushita Shukla and Ajay Arora Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Salicylic acid (SA) is involved in regulation of many processes in plant growth and development such as ripening, abscission, senescence etc. There are many reports in favour of exogenous application of SA acting as a plant growth regulator for enhancing the shelf life of perishables. But, so far no literature is available on endogenous SA in relation to postharvest life. Fruit ripening is a complex process that lead to changes in fruit, increased activity of cell wall-degrading enzymes, increased respiration and ethylene production. Climacteric fruits, harvested at full maturity, can be ripened even after detachment from parent plant. The respiration rate and ethylene formation though minimal at maturity, rise at the onset of ripening, after which it declines. The most studied model of climacteric fruit ripening is tomato. Present study has been carried out on 55 tomato germplasm at different developmental stages. These were further narrowed down to 15 germplasm on the basis of physiological traits. Cell wall degrading enzymes, ethylene evolution and ethylene metabolism associated enzymes were assayed and compared with endogenous SA at various stages of tomato fruit development. Results from this and earlier studies from our lab. suggest that SA play an important role in fruit ripening. Further, molecular mechanism of SA regulating postharvest life in model plant will be elucidated.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 287 Session G : Plant Growth Regulators and Applications
PG341 (IPPC0486) Salicylic acid induced salt tolerance in chickpea (Cicer arietinum L.) R. Dhandapani*, H.P. Vijayakumar, G. Somasundaram, S. Natarajan and S.R. Prasad Plant Physiology, Indian Council of Agricultural Research-Directorate of Seed Research, Mau-275101, India *Presenting author: [email protected]
Salinity is the major abiotic stress affecting germination and early seedling establishment of chickpea. We demonstrated the differential performance of saline sensitive (PG186) and saline tolerant (Karnal chenna) chickpea cultivar under 150 mM Nacl salt stress. Salt stress adversely affected the germination of PG 186 cultivar. Sand matrix priming along with Salicylic Acid (SA) proved to enhance the germination up to 40%. To identify the cellular mechanism of SA-induced salt tolerance, various antioxidant enzymes (AOE) were estimated from cotyledon during germination. Results indicated that, unprimed PG 186 seeds accumulated more AOE than SA-primed seeds. Like SA-primed seeds, unprimed seeds of cultivar Karnal chenna accumulated less AOE in cotyledon and in term balanced the accumulates of more H2O2 signal for better germination. The enhanced H2O2 signal activated several processes including reserve protein mobilisation during germination. Cotyledons of both unprimed and SA-primed seeds were found to contain same level of protein 8 days after were sowing. The initial protein content of SA-primed seeds was more than the unprimed seeds before sowing. These results showed that, SA negatively influenced AOE and in term improved H2O2 signal under high salt stress. SA also induced stress tolerant protein synthesis during first phase of germination and mobilised them during the final phase of germination.
PG342 (IPPC0509) In vitro production of microtuber of potato (Solanum tuberosum L.): Effect of cultivar, sucrose and inorganic nitrogen nutrition Sunil J. Macwan1*, Yogesh M. Shukla2, Natvarlal V. Upadhyay3 and Piyush R. Vaishnav4 1Department of Genetics & Plant Breeding; 2Department of Biochemistry; 3Plant Physiology; 4Department of Agricultural Statistics; B.A. College of Agriculture, Anand Agricultural University, Anand-388110, Gujarat, India *Presenting author: [email protected]
The potato (Solanum tuberosum L.) belongs to the family solanaceae, is a crop of worldwide importance. Two-stage in vitro tuberization process comprising first micro propagation via nodal explants and then tuber induction in the resultant in vitro plantlets was studied using three cultivars of potato, Kufri Chipsona-1, Kufri Badshah and Kufri Pukhrajs. In vitro cultures were established with MS (1962) in absence of growth regulators, prior to surface sterilization with 0.1% HgCl2 for 8 minitues. Axenic cultures were evaluated for maintaining and proliferation of in vitro shoots and concluded that plant growth regulators and nutrients in the medium are key factors for in vitro multiplication of potato micro propagation. Effect of sucrose concentration for induction and in vitro development of micro tuber were studied and concluded that optimum level of sucrose (8%) found to be best for free growth regulators. Further it was noted that stimulation and induction of microtuber initially with full light was essential. Effect of inorganic nitrogen nutrition on the induction and development of microtubers was studied with an objective to investigate whether a reduction in total nitrogen level in the Murashige & Skoog medium (MS, 1962) would improve tuberization rate. It was found that lower level of inorganic nitrogen by nitrate concentration in media gave better response for the microtubrisation. Interaction between cultivar and other factors were found to be significant indicating the importance of developing variety specific protocol to optimize microtubrisation.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 288 Session G : Plant Growth Regulators and Applications
PG343 (IPPC0531) Regeneration of shoot and induction of flowers in Saffron plant (Crocus sativus L.) under in vitro and ex vitro conditions Khirod Sahoo*, Chhavi Sharma and Madhu Sharma Department of Botany, Ravenshaw University, College Sqaure, Cuttack-753003, India *Presenting author: [email protected]
The trifid stigma of Crocus sativus L. flower is valued for the costliest spice of the world-Saffron. Principally, it is used in medicine, colouring and flavouring confectionary items. It is a summer dormant crop and conventionally propagated through corms. The total yield of saffron reduces up to 70% due to various biotic stresses. To develop the genetic transformation method for improvement of saffron quality and quantity, an efficient shoot regeneration protocol is needed. To overcome these issues, we have successfully standardized protocols for breaking the dormancy of corm, multiple bud-sprouting, regeneration of multiple shoots, production of sterile corm and cormlets under in-vitro condition. Importantly, we have standardized a universal protocol to develop multiple shoots from different explants through somatic embryogenesis. This will help to produce pathogen-free germplasms for field level cultivation as well as to develop a genetic transformation method in saffron in future. We have also established methods to induce flower from corms under in-vitro and ex-vitro conditions to understand the hormonal regulation in saffron plants. We found that hormonal regulation for flowering in saffron plant under in-vitro and ex-vitro conditions is partially correlated with each other. The reason for minimal uncorrelation could be due to the difference in environmental conditions. Hence, in-depth molecular study will be needed to comment on the exact hormonal regulation in flower development of saffron plants.
PG344 (IPPC0542) RNAi-mediated knockdown of tobacco ODC gene results in altered plant regeneration, development and stress responses Ami Choubey* and M.V. Rajam Polyamine, Transgenic and RNAi Research Lab, Department of Genetics University of Delhi, South Campus Benito Juarez Marg, New Delhi-110021, India *Presenting author: [email protected]
Polyamines (PAs) are ubiquitously present polycationic compounds that play a critical role in various biological processes. In addition to participating in various growth and developmental processes, PAs are known to elicit stress responses in plants. Yet their specific functions and mode of action remain largely unknown. In plants, complexities intensify due to the existence of two parallel biosynthetic pathways for putrescine (Put) production; one mediated by the universally present ornithine decarboxylase (ODC) and another by arginine decarboxylase (ADC). In the present study, we have targeted ODC by RNA interference to modulate PA levels and study the effects at different developmental time points. Significant physiological and morphological alterations seen in response to ODC knockdown include poor regeneration response from leaf explants, reduced leaf size, reduced chlorophyll and carotene content, increased vulnerability to abiotic stress conditions, delayed flowering, pollen sterility, reduced seed setting, reduced seed viability and delayed seed germination. Also, for the first time, microarray analysis has been attempted in an ODC knockdown line to study genome-wide gene expression changes in response to ODC downregulation and changes in PA titres. On the whole, it presents a detailed study of trying to decipher functions of PAs in tobacco growth and development.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 289 Session G : Plant Growth Regulators and Applications
PG345 (IPPC0573) Tiller dynamics of rice in relation to physiological control of panicle grain yield Kariali Ekamber* School of Life Sciences, Sambalpur University, Jyotivihar, Sambalpur-768019, Odisha, India *Presenting author: [email protected]
Tiller development in rice is asynchronous. During the early growth phase, rice plant produces a large number of tillers in an acropetal fashion form the base upwards. The tillers which originate later are highly unproductive and do not contribute much to the overall grain yield. Leaf longevity, growth duration and pattern of senescence of the flag leaf in a tiller are indicative of competence of the organ for survival and provision of assimilates for grain filling in rice. However, the mechanism by which tiller senescence is induced in the plant for termination of the life cycle is not known hitherto. Therefore, the objective of the present work was to analyze the pattern of senescence, photosynthetic efficiency and lipid peroxidation of the flag leaf in different tiller groups and their impact on grain yield. The results revealed that origin of tillers in the stem node occurred in a temporal fashion. The late-formed tillers had short vegetative period, reduced growth duration, low photosynthetic pigment concentration and higher membrane lipid peroxidation compared to early-formed tillers; the former had limited source activity compared to the latter. Hence, it can be concluded that asynchronous tiller origin might be responsible for production of smaller source area for short growth duration in the late–tillers leading to maintenance of reduced photosynthetic pigments level. High disintegration of membrane lipids leads to the restriction in the primary production of the source to the detriment of grain filling and gain yield in rice.
PG346 (IPPC0595) Improving photosynthetic responses during recovery from heat treatments with brassinosteroid and calcium chloride in Indian bread wheat cultivars Suboot Hairat* and Paramjit Khurana Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India *Presenting author: [email protected]
Climate change is expected to unleash severe and frequent heat waves in future, adversely affecting crop productivity. The aim of this study was to examine the effect of two separate episodes of heat stress, mimicking heat wave conditions on the physiology of two contrasting Indian bread wheat cultivars based on their level of thermotolerance and to study the ameliorating effects of epibrassinolide (BR) and calcium chloride on the recovery of these cultivars. The thermo-tolerant cultivars C306 suffered less inhibition of photosystem II efficiency as compared to the thermo-susceptible cultivars HD2329. Application of BR and calcium chloride resulted in faster recovery in both the cultivars. Measurement of the minimum fluorescence (Fo) versus temperature curves revealed a higher inflection temperature of Fo (Ti) for the tolerant cultivar as compared to the susceptible cultivar, emphasizing greater thermo-stability of the photosynthetic apparatus. The thermo- tolerant cultivar showed higher photochemistry (PSII) relative to the susceptible cultivar. An increase in the steady state fluorescence was observed in the susceptible cultivar HD2329 as compared to the tolerant cultivar C306. Expression analysis revealed faster recovery of the transcripts involved in photosynthesis in tolerant cultivar as compared to the susceptible cultivar. Exogenous application of the ameliorating compounds resulted in faster recovery of transcripts in both the cultivars. The result suggest that under severe stress conditions tolerant cultivar C306 showed faster recovery and a better thermo-stability of its photosynthetic apparatus as compared to susceptible cultivar HD2329 and application of epibrassinolide and calcium chloride can ameliorate the damaging effect of severe temperature.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 290 Session G : Plant Growth Regulators and Applications
PG347 (IPPC0596) Radiation-processed chitosan enhances the crop productivity, essential oil and citral production in lemongrass [Cymbopogon flexuosus (Steud.) Wats.] M. Masroor A. Khan1*, Hassan Jaleel1, Moin Uddin1 and Lalit Varshney2 1Department of Botany, Aligarh Muslim University, Aligarh-202002, India 2 Radiation Technology Development Division, BARC, Mumbai- 400 085, India *Presenting author: [email protected]
Lemongrass is an important medicinal and aromatic plant. Its essential oil (EO) is an excellent source of natural citral, which is used as a raw material for β-ionone synthesis. β-ionone is used for the synthesis of vitamin-A and other compounds. Hence, increased production of lemongrass EO is highly desirable. Recently, a new agro-technique has been evolved in which it has been proved that if natural polysaccharides like alginate, carrageenan and chitosan are exposed to gamma radiation, they are depolymerized into various oligomers. When aqueous solution of these oligomers is applied to the plants, it proved as potent plant growth promoter. Keeping this in mind, a field trial was conducted on lemongrass [Cymbopogon flexuosus (Steud.) Wats.] to study the effect of different concentrations (0, 40, 60, 80 and 100 mg L-1) of irradiated chitosan (ICH) on the growth, biochemical, quality and yield attributes. The foliar sprays were made five times during the crop tenure. The sampling was done at 150 days after planting. The EO was extracted using Clevenger’s apparatus and analyzed using gas chromatography. The effect of ICH was significant on most of the parameters studied, with 80 mg L-1 (ICH-80) proving the best concentration for growth parameters, chlorophyll content, leaf-N, -P, -K cocentrations, carbonic anhydrase activity, nitrate reductase activity as well as EO content and EO yield as compared to control. However, effect of ICH on citral content was insignificant. Conclusively, application of ICH-80 could improve the performance of the crop including EO production of the lemongrass.
PG348 (IPPC0680) High throughput somatic embryogenesis system and stage specific expression of SERK1 gene during somatic embryogenesis in cultures of greengram, Vigna radiata (L.) Wilczek Kumariah Manoharan1*, Vajravel Sindhuja1, Muniraj Gnanaraj1, Nagan Udhayakumar1, Maluventhen Viji2, Sellakumar Maheswari1 and Thirupathi Karuppanapandian3 1Department of Plant Morphology and Algology, School of Biological Sciences, Madurai Kamaraj Univ., Madurai-625021, TN, India 2Department of Botany, Thiagarajar College, Madurai-625009, Tamil Nadu, India 3Department of Experimental Biology, Faculty of Science, Masaryk University, Brno-625 00, Czech Repub *Presenting author: [email protected]
A protocol for attaining high frequency somatic embryogenesis (SE) in green gram (Vigna radiata (L.) Wilczek) was worked out. Type of explants and age of source seedlings for obtaining explants were found to influence the formation of embryogenic callus. Various combinations and concentrations of 2,4-dichlorophenoxyacetic acid and indole-3-acetic acid with kinetin were optimized for developing embryogenic callus. Embryogenic callus when exposed to osmotic stress created by D-mannitol and sorbitol and dehydration stress imposed by polyethylene glycol were found to produce somatic embryos. Callus incubated for 6 h in specified hormone free nutrient medium supplemented with 4% polyethylene glycol was found optimal for the induction of high frequency SE. Subsequent to stress incubation, the cultures formed only early stage somatic embryos with the absence of cotyledonary stage somatic embryos. Supplementation of proline (150 mM) was found essential for the maturation of somatic embryos. The cotyledonary stage somatic embryos were converted into plantlets and subsequently established in garden soil. Semi-quantitative Reverse Transcription-PCR based transcript level analysis of SERK1 gene expression was carried out employing embryogenic callus, callus containing different stages of somatic embryo. Expression of SERK1 was specifically associated with the embryogenic callus and callus enriched with globular stage somatic embryos.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 291 Session G : Plant Growth Regulators and Applications
PG349 (IPPC0708) The role of ethylene biosynthesis inhibitors in modulating in vitro growth of Cicer arietinum seedlings induced by thidiazuron Rachana Dinesh* Department of Botany, Jai Narain Vyas University, Jodhpur-342001, Rajasthan, India *Presenting author: [email protected]
The study aims at critically analyzing the role of ethylene biosynthesis inhibitors in modulating in vitro growth of chickpea seedlings induced by thidiazuron. Since, TDZ has also been implicated in the formation of ethylene hence present investigation was undertaken to evaluate the role of ethylene biosynthesis inhibitors (aminooxy acetate-AOA; aminoethoxyvinyl glycine- AVG; cobalt-CO++) in modulation of the effect of TDZ on growth and hyperhydricity in chickpea seedlings. TDZ inhibited root and shoot elongation and the expansion of leaflets in in-vitro grown Cicer arietinum seedlings. The roots became thick and green, and developed extensive nodular callus. TDZ also promoted prolific formation of shoots from the cotyledonary node, and induced hyperhydricity. Simultaneous addition of inhibitors of ethylene synthesis (AOA, AVG, Co++) together with TDZ resulted in a significant modulation of the effects of the latter chemical on the formation of shoots from the cotyledonary nodes, development of callus on the roots and the induction of hyperhydricity in the seedlings. These results indicate that, to some extent, ethylene may be involved in the morphogenetic responses ilicited by chickpea seedlings in response to TDZ.
PG350 (IPPC0719) Effect of plant growth regulators and nutrients on alleviation of salinity stress effect in tomato physiology and yield R. Sivakumar*, K. Nandhitha and P. Jeyakumar Crop Physiology, Tamil Nadu Agricultural University, Maruthamalai Road, Coimbatore- 641003, Tamil Nadu, India *Presenting author: [email protected]
Agricultural production and soil health is agitated by the accumulation of diverse substances to soil mainly high levels of salts. The main factors that contribute to this problem are the salt load in the water used for irrigation, lack of sufficient rainfall and the use of vast and versatile fertilizers. The soil salinity may cause several deleterious effects on physiological and biochemical parameters, growth and development of crop ultimately yield. Pot culture experiment was conducted to study the physiological changes in tomato genotypes sprayed with PGRs and nutrients under salinity condition (100 mM) in tomato genotypes (PKM 1 and TNAU THCO 3). Foliar application of PGRs like brassinolide (0.5 ppm), salicylic acid
(100 ppm), benzyl amino purine (50 ppm), ascorbic acid (100 ppm), gultathione (50 ppm), KNO3 (0.5%) + FeSO4 (0.3%)
+ Borax (0.2%) and nutrient PGR concoction (K2SO4 (0.5%) + CaSO4 (0.5%) + Borax (0.2%) + NAA (20 ppm) were used at 20 and 40 DAT. Brassinolide recorded highest leaf water potential (-1.51 MPa), maximum relative water content -1 -1 -1 (76.50%), highest soluble protein content (13.53 mg g ), maximum catalase activity (5.48 μg H2O2 g min ), minimum Na/ K ratio (0.43), increasing the leaf magnesium content (0.69) at 60 DAT and superior fruit yield (574.55 g plant-1) followed by salicylic acid and ascorbic acid compared to control. However, brassinolide performed better in yield, salicylic acid recorded highest BC ratio of 3.2 and 2.5 in TNAU THCO 3 and PKM 1 respectively followed by brassinolide (3.1 and 2.3) under salinity.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 292 Session G : Plant Growth Regulators and Applications
PG351 (IPPC0724) De-hulling induced ethylene breaks seed dormancy in wild rice Jayanta Barik*, Ekamber Kariali and Pravat Mohapatra School of Life Sciences, Sambalpur University, Jyoti Vihar, Sambalpur-768019, Odisha, India *Presenting author: [email protected]
Pre-harvest sprouting or viviparous seed germination in rice is one of the major factors causing massive yield losses to rice farmers in many rice growing nations of the world, when climate change-induced erratic rains coincide with grain maturation period. However, many species of wild rice such as Oryza rufipogon and Oryza nivara including some traditional varieties exhibit immunity against viviparous germination in spite of the availability of most suitable environmental conditions. These recalcitrant rice genotypes maintain a fixed dormancy period and germinate only afterwards with the commencement of suitable environmental conditions. This versatile trait is of prime importance for introgression into the non-dormant high yielding rice cultivars. The objective of the present investigation was to induce germination in three different (fully-dormant, semi-dormant and non-dormant) classes of rice seeds by de-hulling their endosperm including the latent embryo to assess the responsible physiological mechanisms. The treatment imposed promoted germination, seed vigour index, ethylene production and mobilisation of soluble assimilates to the culminating sprout. Soluble sugars and starch concentrations were positively correlated with ethylene production. However, ethylene production was negatively correlated with the lag period for seed germination. The results obtained provided evidence for the existence of lemma and palea as a strong physical barrier for germination. Their removal from the seed caryopsis promoted high ethylene production for the access of germination process. De-hulling effects on the genesis of ethylene and its possible role in breaking seed dormancy and related activities has been discussed.
PG352 (IPPC0743) Induction of parthenocarpy in jackfruit (Artocarpus heterophyllus Lam) T.D. Nikam* Department of Botany, Savitribai Phule Pune University, Pune-411007, Maharashtra, India *Presenting author: [email protected]
The jackfruit (Artocarpus heterophyllus Lam) is the largest tree-borne fruit in the world and are excellent source of carbohydrates, minerals, vitamins, fibres, antioxidant and low fat. It is a widely cultivated and popular food item throughout the tropical regions of the world. It is one of the principal fruit trees that offers potential for fruit marketing and income generation in the sector of agroforestry and agriculture.The Jackfruit tree can produce about 100 to 200 fruits in a year. In the present investigation, the female inflorescence at different stages of developement were treated with various concentrations (00, 10, 100, 1000, 10000 ppm) of gibberellic acid, auxins (IAA, NAA, IBA and 2,4-D) and cytokinins (kinetin and BAP). Iduction of parthenocrpic fruit development was observed in 57 treated samples. The morphological, anatomical and biochemical investgations were carried out.Morphological differences were not distinct in parthenocarpic and control fruits but parthenocarpic fruits were lighter in weight. Maturation and ripening period for both remain in the range of 90 to 140 days. Dissection of parthenocarpic fruit revealed that, compared to control, the wall of the fruit was not thick and firm. The pedicel was normal but in parthenocarpic fruits the parianth remains thin. Both types of fruits were characteized for carbohydrate, protein, lipid, minerals, vitamins, and fibre content.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 293 Session G : Plant Growth Regulators and Applications
PG353 (IPPC018)
Elevated CO2 ameliorates drought and high temperature stress effects in rice Madan Pal*, Ashish K. Chaturvedi and Divya Shah Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Apart from rising CO2, high temperature and drought stress are important climate change factors, which are predicted to increase in frequency and magnitude under changing climate. These factors often interact with each other under field conditions and their resulting impacts may be different compared to individual exposure. This work is focused on physiological impact of combinations of these factors compared to independent exposure. The experiments were conducted –1 to analyse the role of elevated CO2 (eCO2, 550±34 μmol mol ) in combination with drought and high temperature stress on growth, gaseous exchange, carbon metabolism, grain yield and quality attributes in Nerica L-44 and Pusa 1121 rice genotypes, raised inside FACE and open top chambers. Rate of photosynthesis, non-structural carbohydrates and
ADPGase and starch synthase enzyme activity was stimulated under eCO2, decreased under drought and high temperature stress and reduction was less under combined exposure of eCO2 with drought or high temperature. Similar response was recorded for dry matter production and grain yield. Chalkiness in grains was more under eCO2 individually and further enhanced under combined exposure of drought and high temperature. Grain protein concentration decreased under eCO2 individually but reduction was less under combined exposure of drought and high temperature with eCO2. Amylose concentration in grains was less under combined exposure of drought and high temperature with eCO2 compared to exposure to eCO2 individually. These findings suggest that negative impacts of drought and high temperature stress on grain yield and quality in rice can be lowered if combined with eCO2 exposure.
PG354 (IPPC0777) Employing some PGRs and elicitors for improved artemisinin production in Artemisia annua L. under abiotic stress M. Masroor Khan, M. Naeem*, Tariq Aftab and Moin Uddin Aligarh Muslim University, Aligarh-202002, Uttar Pradesh, India *Presenting author: [email protected] Artemisia annua L. (family Asteraceae), is the only source of a potent anti-malarial drug, artemisinin. Artemisinin, a sesquiterpene lactone containing an endoperoxide bridge, has been recognized as an effective and safe remedy against malaria parasite Plasmodium falciparum. However, the concentration of artemisinin in the plant is very low, ranging from 0.01 to 0.8%. Exposure of A. annua plants to abiotic stresses, such as of light, temperature, salinity, heavy metals and UV light, have been reported to enhance ROS generation, which augment artemisinin yield by facilitating rapid conversion of artemisinic acid to artemisinin. In fact, generation rate of H2O2 increased consistently under abiotic stress conditions, suggesting a direct relationship between generation of H2O2 and artemisinin biosynthesis in A. annua. Various scientific approaches have been attempted to increase artemisinin production including chemical synthesis and genetic engineering of the genes involved in artemisinin biosynthesis pathway in A. annua; but not much success has been accomplished as yet because of the high cost of procedure or complex nature of regulation and expression of the genes responsible for artemisinin biosynthesis. The PGRs exhibit a broad spectrum of physiological effects in plants, may influence the inter- relationships between primary and secondary metabolism leading to increased biosynthesis of secondary metabolites. A pot and field study employing PGRs (gibberellic acid, salicylic acid and methyl jasmonate) and radiation-processed polysaccharides (sodium alginate and carrageenan) proved as non-conventional elicitors that enhanced the overall yield of artemisinin under normal and abiotic stress conditions. Artemisinin content was analyzed though HPLC. In order to increase the desired production of artemisinin and utilize the land unsuitable for food crops, cultivation of A. annua might be employed with new scientific approaches and convenient strategies.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 294 Session G : Plant Growth Regulators and Applications
PG355 (IPPC0824) Elucidating the role of strobilurin in combination with tebuconazole on physiology of crops Jeyakumar Prabhakaran* and Jidhu Vaishnavi Sivaprakasam Crop Physiology, Tamil Nadu Agricultural University, Maruthamalai Road, Coimbatore- 641003, Tamil Nadu, India *Presenting author: [email protected]
The new generation fungicides have been found to have phytotonic effects in many crops. Strobilurin fungicides can supplement nitrogen demand of plants by strongly activating nitrate reductase and in turn nitrate assimilation. In the present study, strobilurin (trifloxystrobin) in combination with triazole is reported to exhibit significant physiological and biochemical changes in groundnut, cotton and chillies. Foliar spray of Nativo (trifloxystrobin + tebuconazole) at different concentrations was taken up at different growth stages of crops. Chlorophyll index, photosynthetic rate, nitrate reductase activity, nitrogen uptake by the plants and their content in soil were recorded at selected stages up to harvest. Nativo @ 300-400 g ha-1 was found effective in enhancing the yield of crops due to changes in physiological characters studied. The combined application of trifloxystrobin and tebuconazole enhanced the ‘greenness’ of the crop by showing higher chlorophyll index with an increase of more than nine per cent compared to untreated control. The improvement in ‘greenness’ can be ascribed to higher uptake of nitrogen by the crops and efficient nitrate reductase activity. The nitrogen uptake and nitrate reductase activity were found higher by more than 10 and 14% compared to untreated control. The better nitrogen assimilation favoured for enhanced photosynthetic efficiency. The photosynthetic rate was significantly higher by recording -2 -1 -1 39.79 and 35.92 μmol CO2 m s in groundnut and chillies. In cotton, higher soluble protein content (10.57 mg g ) favoured for enhanced photosynthesis. The combined effect of strobilurin and triazole thus enhanced physiological efficiency and in turn.
PG356 (IPPC0827) Efficacy of plantozyme on physiological parameters and yield components in thompson seedless grapes under pune condition S.D. Ramteke* ICAR-National Research Centre for Grapes, Pune-412307, Maharashtra, India *Presenting author: [email protected]
An investigation was carried out to study the efficacy of Plantozyme on physiological parameters, quality and yield components in Thompson Seedless grape at NRC Grapes, Pune during 2012-13, 2013-14 and 2014-15. Eleven treatments were applied in replicated RBD experiment. Morphological and biochemical parameters were recorded at 90 days after pruning and yield parameters were recorded at harvest. The results revealed greater values of morphological characters, i.e., shoot length (138.8 cm) and leaf area (243.8 cm2) and quality parameters like berry diameter (17.62 mm), berry skin thickness (30.94 μ), and berry length (21.62 mm) in treatment T8, while cane thickness (8 mm), leaf thickness (0.22 mm), biochemical parameters such as reducing sugar (3.75 mg/g), total protein (2.25 mg/g) and total phenol (2.25mg/g) were higher in T9. The highest yield per vine (16.01 kg) and BC ratio (3.24) were obtained in T10. This study concluded that plantozyme increased berry characters, quality parameters and biochemical parameters as compared to control. Photosynthetic parameters were also influenced with the application of Plantozyme. Application of Plantozyme through soil @ 50 g/ vine, drip @ 2.5 ml/ vine and spray @ 2 ml/ l improved overall quality and yield of Thompson Seedless grapes under Pune conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 295 Session G : Plant Growth Regulators and Applications
PG357 (IPPC0859) Effect of micronutrient and growth regulators through foliar application on chickpea and wheat under late sown conditions S.R. Kushwaha*, C. Viswanathan, Chandrapal, Rakesh Verma, Suchitra Puskar and D.V. Singh Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Foliar nutrition is known to correct the nutrient deficiencies usually observed at seed development in crops. A field trials was conducted at Indian Agricultural Research Institute, New Delhi to find out effectiveness and feasibility of different nutrient combination and growth regulators applied through foliar spary at 50% flowering in chickpea and wheat. Chickpea cultivar Pusa 362 and wheat HD 2967 were grown in field condition under late condition during 2013-14 and 2014-15. Observation was recorded on physiological traits like chlorophyll content, membrane stability index (MSI), yield and yield attributes. Results showed that physiological parameters i.e. RWC, MSI, chlorophyll content were significantly enhanced (12-22%) by the foliar application of micronutrient combination. The combination of micronutrient increased yield over control by 39.34% while application of growth regulators the yield was improved by 18 to 22% under late planting conditions. The yield attributes of chickpea and wheat were also increased by micronutrient combination and growth regulators over the control under late sown conditions.
PG358 (IPPC0870) Role of glucose in regulating directional growth responses in Arabidopsis Manjul Singh*, Aditi Gupta and Ashverya Laxmi National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Phytohormones and sugars/glucose (Glc) affect a number of developmental, physiological and metabolic processes in plants. Here, we try to understand the role of glucose and its interaction with phytohormones in controlling root directional responses. Presence of Glc in the medium could extensively modulate seedling root growth direction. Whole genome transcription profiling revealed that in Arabidopsis roots growing in sugar free medium, gravity reorientation could differentially regulate a significant number of genes. Whereas, a completely different set of genes were found to be affected upon gravity reorientation in presence of Glc. Interestingly, Glc alone could also affect a large number of these gravity reorientation regulated genes under unstimulated condition. The effects of Glc upon transcript levels of gravity- reorientation specific genes were largely antagonistic. These results suggested the existence of a tightly regulated genetic network of Glc modulated gravitropic response in roots. Glc could also regulate several phytohormone related genes previously known to be involved in gravitropic responses even in unstimulated condition indicating Glc and hormone interplay during gravitropic growth. BR signaling works downstream to Glc in regulating root directional growth. Glc could also induce BR receptor endocytosis. Mutant phenotypes as well as pharmacological assays suggested an important role for protein dephosphorylation during Glc-BR induced root-deviation response. Auxin signaling and polar transport components are also involved in Glc induced changes in root directional growth. Altogether, Glc mediated change in root direction is an adaptive feature which is a result of a collaborative effort integrating phytohormonal signaling cues.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 296 Session G : Plant Growth Regulators and Applications
PG359 (IPPC0882) Hormonal impacts on physiological processes of rice in relation to methane emission Leena Borah* and K.K. Baruah Department of Environmental Science, Tezpur University, Tezpur-784028, Assam, India *Presenting author: [email protected]
Rice paddies are major sources of the non-CO2 greenhouse gas methane (CH4), which contributes significantly to global warming and climate change. The rice plant mediates the transport of CH4 from soil to the atmosphere. Therefore, physiological and anatomical characteristics of the rice plant play a regulatory role in CH4 transport. Impact of exogenous application of growth hormones viz. gibberellic acid (GA), indole acetic acid (IAA), indole butyric acid (IBA) and kinetin -1 (KIN) (@ 20 mg L ) on physiological and anatomical characteristics in relation to CH4 emission from rice paddy (variety Luit) was worked out in a two-year field study. IAA and KIN application significantly enhanced the rate of flag leaf photosynthesis in the rice plants over untreated (control) plants. The rate of flag leaf transpiration was lower in IAA and KIN treated plants in comparison to untreated plants which might be attributed to the lower stomatal frequency in the flag leaf and smaller xylem vessel diameter of the topmost node observed in hormone treated plants. IAA and KIN applications were found to bring about a reduction in cumulative CH4 emission by ~10% over control. Remarkable improvement of grain yield and yield attributing characteristics were observed due to application of auxin (IAA) and kinetin. Physiological manipulations in rice plant by growth hormone application can serve as an effective strategy for improvement of rice productivity and simultaneous mitigation of CH4 emissions.
PG360 (IPPC0933) Physiological and biochemical effects of 24-epibrassinolide on heat stress adaptation in maize (Zea mays L.) Jyoti Kaushal1, Anuradha Gautam2, Hemangini Parmar2, Ishwar Singh2 and Pranjal Yadava2* 1Sam Higginbottom Institute of Agriculture, Technology & Sciences, Allahabad-211007, Uttar Pradesh, India 2ICAR-Indian Institute of Maize Research, Pusa Campus, New Delhi-110012, India *Presenting author: [email protected]
Climate change and associated extreme weather events have made high temperature a major threat to crop production worldwide. Brassinosteroids (BRs) are a new type of polyhydroxy steroidal phytohormones that regulate a number of physiological processes in plants. Among these, Brassinolide (BL), 28-homobrassinolide (28-homoBL) and 24- epibrassinolide (24-epiBL) are commonly used. The present study aims at studying the usefulness of 24-epiBL in ameliorating the negative impacts of heat-stress in maize along with its role in regulating cellular antioxidant defence system. PMH 3 maize hybrids were grown in pots in green house maintained at 14 hours day (25oC) /10 hours night (17oC). 24-epiBL solution (1 μM) was applied externally at V4 stage. Leaf tissues were sampled from both treated and control plants. Subsequently, both the groups of pots were placed in plant growth chamber maintained at high temperature (48oC; RH 50%). Plants were sampled for biochemical analysis after 3, 6, 9, 24 and 48 hours of high temperature exposure. Exogenous application of 24-epiBL arrested protein degradation and enhanced membrane stability, as compared to the control. The activity of antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT) was also found to be increased post 24-epiBL treatment. Thus, the study supports the role of BRs as anti-stress agents. However, investigations are required to gain further insights into the molecular-genetic mechanisms of BRs action in modulating the plant defence system under different stresses.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 297 Session G : Plant Growth Regulators and Applications
PG361 (IPPC0971) Depolymerised sodium alginate and methyl jasmonate improve the growth, physiological attributes and alkaloids production in Catharanthus roseus (L.) G. Don. Moin Uddin*, M. Masroor A. Khan and Lalit Varshney Aligarh Muslim University, Aligarh-202002, Uttar Pradesh, India *Presenting author: [email protected]
Irradiated sodium alginate (ISA) as well as methyl jasmonate (MeJ) acts as endogenous growth elicitors, triggering the synthesis of different enzymes and modulating various plant responses by exploiting the gene expression. Two pot experiments were conducted to test whether the foliar application of ISA and that of ISA+MeJ could augment the active constituents as well as growth, physiological and yield attributes of Catharanthus roseus, which carries anticancer alkaloids (vincristine and vinblastine) in its leaves in addition to various other useful alkaloids. Totally, 5 spray treatments, comprising various aqueous solutions of ISA [20, 40, 80 and 160 mg L-1 (Experiment 1)], and those of ISA+MeJ [40+20, 40+30, 80+20, 80+30, 160+20 and 160+30 mg L-1 (Experiment 2)], were applied at seven days interval. Leaf-alkaloids content and growth, physiological and yield parameters, evaluated at 120 days after sowing, were significantly improved by ISA application. ISA application could not increase the leaf-content of vincristine and vinblastine; however, it significantly augmented the yield of these alkaloids because of enhancing leaf dry matter. Compared to the control (water-spray treatment), collective application of ISA (80 mg L-1) and MeJ (20 mg L-1) resulted in the highest values of most parameters studied. However, 80 mg L-1 of ISA applied with 30 mg L-1 of MeJ maximally improved the content and yield of total as well as anticancer leaf-alkaloids. Comparing the control, it increased the content and yield of total leaf-alkaloids (30 and 73%, respectively) and those of vincristine (55 and 134%, respectively) and vinblastine (19 and 79%, respectively).
PG362 (IPPC1013) Artificial seed production of Gentiana kurroo Royle Payal Kotvi*, EneshVashist, Shivam Sharma and Hemant Sood Department of Biotechnology and Bioinformatics, Jaypee University of Information and Tech. Waknaghat, Solan-173234, HP, India *Presenting author: [email protected]
Gentiana kurroo royle is a critically endangered plant, which is known to be a bitter drug plant in Indian medicine. It has been widely distributed in places like Jammu and Kashmir, Himachal Pradesh, in northern western Himalayas between the altitude ranges of 1500 and 3500 m, Nepal and Pakistan. It has been confirmed to be the major producer of secondary metabolites belonging to the family Gentianaceae. In this study somatic embryogenesis was conducted as a source of artificial seed production of Gentiana kurroo royle. Artificial seed production has led to increase in the viability rates and germination capabilities of Gentiana kurroo royle seeds in soil, thereby providing efficient means of germplasm conservation, easy transportation and large scale propagation of endangered species. Morphogenetic capabilities of leaf explants have been optimised on MS media containing different concentrations and combination of KN+2, 4-D and out of these KN 1 mg l-1 and 2,4-D 0.5 mg l-1 gave best results for callus initiation and proliferation. Once the callus was induced torpedo shaped somatic embryos were selected for artificial seed production. The artificial seed were produced using the sodium alginate (2%) and calcium chloride solution (5%), which led to calcium alginate bead formation, which were tested for germination under in vitro conditions for direct shoot formation. Hence this technique would contribute in protecting the endangered plant species of Gentiana kurroo royle in short duration of time and provide platform for production of important medicinal compounds.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 298 Session G : Plant Growth Regulators and Applications
PG363 (IPPC1025) Transcriptomic analysis of two differentially ripening mango varieties reveals differences in gene expression of major pathways Smriti Srivastava*, Garima Pathak, Ridhi Goel, M.H. Asif and V.A. Sane Plant Gene Expression Lab, CSIR-National Botanical Research Institute, Lucknow-226001, India *Presenting author: [email protected]
Ripening of fruit, known to be governed by plant hormone ethylene (in case of climacteric fruits), involves a complex interplay of several ripening related genes known to be involved in ethylene biosynthesis and its signaling along with genes related to cell wall hydrolysis. Mango, a typical climacteric fruit ripens with a burst of ethylene. There are more than 1000 varieties of mango differing in their ripening pattern. Dashehari (and many other varieties) shows a characteristic pattern, wherein ripening and softening is initiated from the stone and moves towards the periphery. In contrast, ripening in Banganpalli (south Indian variety) is initiated at the periphery and moves towards the stone. In an attempt to decipher the molecular basis of these differences in ripening pattern between the two varieties along with complex spatio-temporal control of ripening, transcriptomic studies were carried out using hybrid assembly of 454 pyrosequencing and Illumina. From this we obtained 86,083 transcripts. Out of these approx 200 were ripening related. Detailed analysis revealed large scale changes in gene expression during the course of ripening between the two varieties. Which includes differential expression patterns of several ACSs and ACOs along with cell wall hydrolases and some aroma and flavour related genes. Detailed studies are in progress to know regulators which cause these changes.
PG364 (IPPC1026) Understanding the role of gluconokinase in regulating developmental flower senescence Sadaf Khan*, Abdul Azeez, Pravendra Nath and A.P. Sane Plant Gene Expression Lab, CSIR-National Botanical Research Institute, Lucknow-226001, India *Presenting author: [email protected]
Gladiolus is a popular garden ornamental, however, the individual flowers of a gladiolus spike suffer from a short vase life, which is undesirable in floriculture industry. We had previously identified a gluconokinase gene that was up-regulated during floral senescence in Gladiolus. The enzyme gluconokinase is a component of the pentose phosphate pathway, where it catalyzes the phosphorylation of gluconate. But there is very little information regarding the role of gluconokinase in plants. This study was carried out to investigate the role of gluconokinase in floral senescence. Gluconokinase activity was found to progressively increase during the course of floral senescence starting from stage 0 (open flower) onwards, concomitantly with increase in transcript level. Maximum activity was observed during late senescence at stage 2. The gluconate level exhibited a peak at stage 1, whereas the 6-phosphogluconate level showed no such stage dependent variation. Feeding gluconate to excised leaves and flowers induced petal and leaf wilting and loss of structure within 36 h. A decrease in Fv/Fm values was observed which could be attributed to the decrease in chlorophyll content upon gluconate treatment. No such change in Fv/Fm and chlorophyll content was observed in flowers and leaves treated with water or with an equivalent concentration of glucose. These studies provide evidence for a hitherto unknown role for gluconokinase-gluconate in senescence in plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 299 Session G : Plant Growth Regulators and Applications
PG365 (IPPC1027) Hormonal and genetic control of root development in tomato Vinod Kumar*, Deepika Singh, A.P. Sane, V.A. Sane and C.S. Nautiyal Plant Gene Expression Lab, CSIR-National Botanical Research Institute, Lucknow-226001, India *Presenting author: [email protected]
Root development is a complex process governed by a fine balance of different hormones that regulate primary and lateral roots differently. To obtain an insight into the factors that govern root development in tomato, a differential transcriptome from tissues such as root, stem, leaves, fruit and flowers was first developed to identify genes that are primarily expressed in root tissues. Through bioinformatic analysis 279 genes that were predominantly expressed in root samples at levels 3-10 fold higher than the rest of the tissues were identified. These included about 92 genes that encoded transcription factors and regulatory proteins. Some of the genes encoding members of the ERF family, Myb domain family, START domain family and WRKY family were chosen for further studies. Expression analysis at different stages of root development confirmed the high expression of these genes primarily in the root tissue. Full length cDNAs and promoters of these have been isolated for further studies. Simultaneously, a hydroponic system for root analysis was developed. The effect of different hormones such as auxin, ABA, ethylene, cytokinin and some of their inhibitors was studied on primary and lateral root development.
PG366 (IPPC1028) Petal abscission in fragrant and non-fragrant roses is associated with differential expression of ethylene biosynthesis and signalling genes Priya Singh*, Amar Pal Singh and A.P. Sane Plant Gene Expression Lab, CSIR-National Botanical Research Institute (CSIR), Lucknow-226001, India *Presenting author: [email protected]
Organ abscission is an important developmental process governed by ethylene. In rose, abscission in the fragrant variety R. bourboniana is highly sensitive to ethylene, while the hybrid variety, R. hybrida shows greatly reduced sensitivity. To gain an insight into ethylene responses during petal abscission, the expression of several ethylene biosynthesis/receptor/ signalling genes was studied during the course of abscission in fragrant and non-fragrant roses. Transcript accumulation of most ethylene biosynthesis genes increased rapidly in petal abscission zones of R. bourboniana within 4-8 h of ethylene treatment, followed by a decrease. A similar kinetics of transcript accumulation was also observed for most receptor and signalling genes. Under field conditions where abscission is delayed, a delay in peak transcript accumulation could be observed in most ethylene biosynthesis/signal transduction genes. Surprisingly, transcript accumulation of most ethylene biosynthesis/signalling genes decreased considerably in ethylene treated petal abscission zones of R. hybrid, with the exception of ACS1 and EIN3. These results suggest that the activation of the ethylene biosynthesis /signal pathway is most likely responsible for the rapid abscission of petals seen in Rosa bourboniana. In contrast its suppression in R. hybrida (except for ETR2, ACS1 and EIN3) through a hitherto unknown mechanism might be responsible for the delayed abscission in this species.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 300 Session G : Plant Growth Regulators and Applications
PG367 (IPPC1032) A negative regulator of ABA responses governs fruit ripening in tomato Asmita Gupta*, Rakesh Upadhyay, Rashmi Garg, Pravendra Nath and A.P. Sane Plant Gene Expression Lab, CSIR-National Botanical Research Institute, Lucknow-226001, India *Presenting author: [email protected]
Abscisic acid (ABA) is a plant hormone, which regulates seed dormancy, plant growth, and responses to environmental stresses. In recent years, a role for ABA in regulating fruit ripening has also been indicated. We have previously identified SlERF6 as a ripening related AP2/ERF domain gene in tomato. In order to get an insight into its function lines over- expressing and under expressing SlERF6 were generated. SlERF6 over-expressors showed early seed germination, reduced ABA inhibition of root growth and reduced stomatal closure in response to water stress, suggesting reduced ABA sensitivity conferred by SlERF6. Expression of SlERF6 is suppressed by ABA. Manipulation of its expression also affected fruit ripening and fruit size, with delayed ripening and reduced fruit size being observed in over-expressors and early ripening being seen in antisense lines. The shift in ripening by about 5 days was most prominent in the breaker stages of sense and antisense lines. The change in expression of SlERF6 was associated with changes in expression of some signaling related genes as well as SlCYP707A3. Analysis of expression of SlERF6 in ABA biosynthesis mutants like notabilis and flacca revealed much higher transcript levels at the immature green, mature green and breaker stages of their fruit compared to Ailsa Craig fruits. The results suggest a crosstalk between ABA and ethylene during fruit ripening that might be mediated by SlERF6.
PG368 (IPPC1036) Early wound induction responses in chickpea are partly controlled by JA dependent and JA independent manner S.P. Pandey, Shruti Srivastava* and A.P. Sane Plant Gene Expression Lab, CSIR-National Botanical Research Institute, Lucknow-226001, India *Presenting author: [email protected]
Chickpea (Cicer arietinum) is an important pulse crop in India and other Asian countries that suffers from severe yield losses due to insect attack from pod borers such as H. armigera. To prevent losses from insects, a detailed understanding about early wound responses is required. Towards this aim, a comparative analysis of transcriptomes of unwounded and insect saliva treated and mechanically wounded chickpea leaves was performed. About 10% of the transcriptome was found to be differentially expressed upon wounding. In silico analysis showed the differential expression of several kinases, phosphatases and transcription factors belonging to the WRKY, MYB, ERFs, bZIP families etc. Genes encoding components of jasmonic acid and ethylene pathways were up-regulated, while growth associated hormone pathways like auxin were suppressed. Validation of expression of several genes was performed and revealed a similar trend of transcript abundance over a detailed time course study as in transcriptome sequencing analyses, indicating that changes determined by transcriptome sequencing were true to nature. The expression of these genes was also tested in response to JA and SA treatments and revealed that not all genes were activated by JA and SA. Moreover, even those that were activated showed a time delay as well as reduced intensity at peak activation compared to mechanical wounding. These results suggest that early wound responses are partly dependent but largely independent of jasmonic acid.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 301 Session G : Plant Growth Regulators and Applications
PG369 (IPPC1041) Biochemical basis of resistance to Alternaria leaf spot in groundnut breeding lines Narendra Kumar*, Nidhi Radadiya, M.K. Mahatma, K.S. Jadon, A.L. Rathanakumar M.C. Dagla, S.K. Bishi, Lokesh Kumar and B.M. Chikani ICAR-Directorate of Groundnut Research, P.B.No.5, Ivnagar road, Junagadh-362 001, Gujarat, India *Presenting author: [email protected]
Groundnut (Arachis hypogaea L.) is an important food legume crop. Several biotic stresses limit groundnut productivity, among which the Alternaria leaf spot (ALS) is becoming a major disease of groundnut in rabi-summer. To understand the biochemical basis of resistance, few advanced groundnut breeding lines showing resistance and susceptible to ALS were screened during summer-2015. Disease severity was assessed using a scale of 1 to 9, where 1 (no symptoms) represents resistant and 9 is 81% to 100% defoliation represents susceptible. The identified lines were characterized by assaying the induction of polyphenol oxidase (PPO), peroxidase (POX) and phenol profiles. Leaf samples were collected at 75 days after sowing from field. Breeding lines showing higher disease incidence (PBS-18057) had minimum activities of PPO and POX enzymes and levels of coumaric and salicylic acid. On the contrary, breeding lines with less disease incidence had higher activities of PPO and POX enzymes along with higher contents of coumaric and salicylic acid (about 2-3 fold). As higher induction of PPO, POX, coumaric and salicylic acid were negatively correlated with disease severity. Hence these parameters can be used as biochemical markers for screening groundnut genotypes resistant to Alternaria leaf spot in future breeding programmes.
PG370 (IPPC1043) Application of paclobutrazol to ameliorate water deficit stress effects in chickpea Soumya P.R.*, Pramod Kumar and Madan Pal Singh Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Chickpea is an important winter season pulse crop of arid and semi arid regions and is sensitive to water deficit stress. A study was conducted to analyse role of Paclobutrzol to ameliorate water deficit stress effects in chickpea. Experiment was conducted using two contrasting chickpea varieties viz. desi (Pusa 362) and kabuli (Pusa1108). There were two main treatments (i.e. well watered and water deficit condition) and four sub-treatments (PBZ foliar spray (60 ppm), PBZ drenching (120 ppm), water spray and water drenching). Optimum dose of PBZ for foliar and drenching was identified based on preliminary study. The results showed decreased RWC, MSI, rate of photosynthesis, Total chlorophylls and carotenoids, grain yield and disrupted chloroplast grana under imposed water deficit stress. However, application of PBZ in general, maintained higher values of all above parameters and maintained the integrity of chloroplast structure under imposed water stress and also stimulated faster recovery after stress termination in both the varieties. PBZ reduced the lipid peroxidation and membrane leakage. It enhanced the activity of antioxidant enzymes and improved stress tolerance ability in both chickpea varieties. In general PBZ drenching was found better over the foliar application. The findings depict that PBZ enhanced the yield by maintaining RWC, MSI, photosynthetic activity, photosynthetic pigments and protected the photosynthetic machinery by restricting lipid peroxidation, increasing antioxidant activity and ABA level, thereby enhanced water deficit stress tolerance.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 302 Session H : Photosynthesis
PH371 (IPPC0132) Chlorophyll a fluorescence changes in response to short and long term high light stress in rice seedlings P. Faseela and Jos Puthur* Department of Botany, University of Calicut, Calicut-673635, Kerala, India *Presenting author: [email protected]
An investigations was carried out to study the high light stress effects on photosystem (PS) II efficiency in intact leaves of four rice (Oryza sativa L.) varieties viz. Jyothi, Aiswarya, Swarnaprabha and Kanchana. Rice seedlings were exposed to high light stress [2,000 μmol (photon) m-2 s-1] and the photochemistry of the four rice varieties was studied by the application of qualitative and quantitative analysis of chlorophyll (Chl) a fluorescence at 0 (control), 2 and 8 h. The results showed that some Chl a fluorescence parameters were highly affected under high light stress. It was found that Fv/Fo, performance index, area above the ûourescence curve, φo/(1- φo), φDo, φEo and Vj are indicators found to be more reliable for exploring the effect of changes in PSII activity after high light stress application for short term (2 h), which can aid in distinguishing the tolerant and sensitive rice varieties. Some Chl a fluorescence parameters were modified only after exposing the plants to high light stress for 8 h. The decline in photochemistry was more pronounced in Kanchana and lower in Jyothi as compared to other varieties and the order of high light stress tolerance of these varieties on the basis of Chl a fluorescence parameters was Jyothi > Aiswarya > Swarnaprabha > Kanchana.
PH372 (IPPC0347) Enhanced photosynthesis is associated with efficient source-sink interaction and nitrogen use in Jatropha curcas L. under elevated CO2 Sumit Kumar* and Attipalli Reddy Department of Plant Sciences, University of Hyderabad, Hyderabad- 500046, Andhra Pradesh, India *Presenting author: [email protected]
The present work demonstrates leaf gas exchange characteristics and related morphological and biochemical parameters in Jatropha curcas on a potential biofuel plant exposed to elevated [CO2] (~550 ppm). Light saturated net photosynthetic rates of upper-canopy leaves were 45% higher under elevated [CO2] compared with ambient [CO2] over the 3-yr period in spite of reduced stomatal conductance (gs) and decreased transpiration rate (E). There were no significant CO2 treatment effects on maximum photosynthetic (Amax) or biochemical capacity (in vivo rate of RuBP carboxylation, Vcmax or in vivo rate of electron transport driving RuBP regeneration, Jmax) of J. curcas leaves as evidenced from A/Ci curve analysis. Chlorophyll a fluorescence measurements, indicated by Fv/Fm and ÄF/Fm ratios and photosystem-II associated electron transport rate (ETRmax), positively correlated with enhanced photosynthesis and growth of the plants in elevated [CO2] conditions. Despite increased area-based leaf sugar (13%) and starch content (25%), and reduced mass and area-based leaf nitrogen concentration (NM, 10%; NA, 6%), the plants showed better photosynthetic nitrogen use efficiency (PNUE) in elevated CO2 environment. The study also revealed that CO2 enrichment significantly influenced female to male flower ratio, fruit and seed yield, above ground biomass and carbon sequestration potential (24 kg carbon per tree per year) in
Jatropha. Our data demonstrate that J. curcas was able to sustain enhanced rate of photosynthesis in elevated CO2 conditions as it had sufficient sink strength to balance the increased biomass yields.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 303 Session H : Photosynthesis
PH373 (IPPC0373) The comparative analysis of PSII protein profiles of some plants, grown under enhanced solar UV-B (280-320 nm) radiation N. Shanthi1* and S. Jannita Nithia 1Department of Botany, Pachaiyappa’s College, Chennai, Tamil Nadu, India 2Department of Botany, Sri Meenakshi Government College for Women, Madurai, Tamil Nadu, India *Presenting author: [email protected] The ozone layer depletion and UV-B radiation fluctuation in the earth’s surface is make great attention today world. The plants like Black gram, Green gram .Radish, Carrot was growing in field condition under enhanced UV-B radiation. Like tropical plants the temperate plants also successfully grown in tropical region. The UVB radiation affects growth, Development, photosynthetic activity of plants. In photosynthetic protein the PSII is the main target for UV-B radiation. Enhanced UV-B radiation leads to a change in the organization of PSII complex. It specifically damaged the D1 polypeptide. The UVB radiation damage and affect PSII protein expression in sensitive plant. In Black gram and green gram the UVB radiation did not produce any significant difference in the PSII polypeptides as compared to other plant. Where as in Radish the UV-B radiation increase accumulation of 33-28kDa protein and affect express the 68kDa protein. In the case of Carrot the large degradation of 32 and 33 kDa, respectively D1 and D2 protein were observed. This severe sensitivity of the D1 protein is undoubtedly a major contributor to the loss of PSII function on exposure to UV-B radiation. From this study, we confirmed the tropical plant naturally resistant to UVB radiation and restore its activity from damage. Whereas Carrot plant more sensitive to UVB radiation the fail to restore it activity from damage. In Radish moderately resistant to UVB radiation they try to stabilize its activity from damage.
PH374 (IPPC0375) Drought reduces photochemical efficiency and gene expression of photosynthesis JoãoAraújo Jr, Thayssa Schley* and Luiz Fernando Almeida State University of São Paulo - Campus Botucatu, Brazil *Presenting author: [email protected]
Current climate changes have resulted in water scarcity in agriculture. Drought interferes in crops productivity. It affects food demand for human population. Therefore, our objective was to evaluate drought effects on physiological and molecular responses of sorghum (Sorghum bicolor L.). For this, one group of plants were subjected to daily watering (control), while in other group watering was withheld. We determined last day of experiment when photochemical process and stomatal conductance (gs) variables showed severe stress. During water deficit stress (29 days), plants showed decline in gs and leaf water potential as compared to control. It indicates sorghum has stomatal closure and osmotic adjustment to prevent water loss. Stress also reduced photochemical dissipation by 50.4% and heat dissipation in reaction centers by 37.1 %, and increased heat dissipation in antenna by 40.2%. It indicates prejudice in photochemical phase and activation in photo-protection process. We also compared mRNA differentially expressed between dry and irrigated conditions. In stressed group, preliminary results of transcriptome analysis indicate increased expression of genes ERD10, HSP70 and Tpt_PEP_trans_dom related to cell protection under dehydration. Stressed group also had greatly reduced expression in genes involved in photosynthesis, viz., LHB1B2 and LHCB4 genes involved in antenna structure and RBC51A (rubisco enzyme). Detected physiological changes are validated by differential expression of genes related mainly to photosynthetic process and drought tolerance. Drought decreased plant productivity not only by stomatal restrictions but also by inducing changes in gene associated with photosynthetic activity. These results help to understand how sorghum tolerates drought and damages that stress can cause in species.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 304 Session H : Photosynthesis
PH375 (IPPC0379)
Differential sensitivity to nitric oxide (NO) and hydrogen peroxide (H2O2) of photosynthesis and respiration in mesophyll cell protoplasts of pea Sunil Bobba* and A.S. Raghavendra Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, Andhra Pradesh, India *Presenting author: [email protected]
The direct effects of nitric oxide (NO) and reactive oxygen species (H2O2) on photosynthesis and respiration in leaf tissues are debatable. We studied the consequences of exogenous application of NO (in the form of SNP) and H2O2 on the patterns of photosynthesis and respiration in mesophyll protoplasts of pea plants. Increased levels of NO severely inhibited the photosynthesis of mesophyll protoplasts, while exhibiting marginal or no effect on dark respiration. The inhibition of photosynthesis by SNP was completely reversed by cPTIO, a NO specific scavenger. In contrast, externally added H2O2 resulted in drastic decrease in the rates of respiration, with only a marginal effect on photosynthesis. At the concentration used in our study, H2O2 didn’t affect the electron transport activities of PS I or PS II. However, both these photochemical reactions showed marked sensitivity to NO in cells. Our observations suggest that chloroplasts could be primary targets of NO and mitochondria for ROS in plant cells. We also attempted to assess the role of crosstalk between chloroplast and mitochondria, during the effects of NO and H2O2. Modulation of NO did not alter much the sensitivity of photosynthesis in mesophyll protoplasts, to antimycin A or salicylhydroxamic acid (SHAM). On the other hand, the presence of H2O2 decreased the sensitivity of photosynthesis to mitochondrial inhibitors, particularly SHAM, suggesting the role of alternative oxidase pathway (AOX) during H2O2 exposure. It appears that H2O2 could mediate the crosstalk between chloroplasts and mitochondria.
PH376 (IPPC0410) Photosynthesis, dry matter partitioning and grain yield in relation to methane emission in a rice (Oryza sativa) ecosystem Ashmita Bharali*, Kushal Kumar Baruah and Sunitee Gohain Baruah Department of Environmental Science, Tezpur Central University, Napaam-784028, Assam, India *Presenting author: [email protected]
Methane (CH4) is second most potent green house gas in the atmosphere after carbon dioxide (CO2). Rice paddies are the largest anthropogenic methane source and produce 7–17% of atmospheric CH4. A field experiment was conducted at Tezpur University during monsoon rice ecosystem (from June-November, 2014), to assess the integrated effects of organic amendments on methane emission and its relationship with photosynthesis, dry matter partitioning and grain productivity. Soil organic amendments resulted in differential methane emission from the rice field and had significant impact on leaf photosynthesis in rice. Flag leaf photosynthesis revealed a strong association with yield and had an inverse relationship with CH4 flux. There was a good correlation between the photosynthesis and sucrose-phosphate synthase (SPS) activity of the flag leaves. Leaf SPS activity increased with the proportion of dry matter in leaves and was inversely related with the dry matter of the roots and cumulative CH4 emission. Application of Azolla compost resulted in superior yield attributing parameters along with higher flag leaf photosynthesis over other treatments. SPS activity in the leaves plays a pivotal role in carbon partitioning and therefore high SPS levels boosted photosynthetic rates under favorable conditions and may indirectly contribute to reduction in CH4 emission.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 305 Session H : Photosynthesis
PH377 (IPPC0421) Screening of mango (Mangifera indica L.) varieties for photosynthetic light response curve in Konkan region of India Mahesh Kulkarni*, Murad Burondkar, Parag Haldankar, Kuruva Mallikarjuna and Kaustubh Kelkar Department of Horticulture, Dr. Balasaheb Sawant Konkan Agriculture University, Dapoli-415712, Maharashtra, India *Presenting author: [email protected]
Konkan region on west coast of India, is one of the biggest (0.182 million ha.) mango growing belts in India. Heavy rainfall (3500-4000 mm), low sun light during monsoon period (June to September) are considered as the major environmental factors, limiting the carbon assimilation and yield. An experiment was conducted during 2014 to screen the commercially important nine mango cultivars of the Konkan for their photosynthetic response at different levels of light intensity i.e. 100, 250, 400, 500, 600, 700, 800, 1000, 1250, 1500, 1750, 2000 μmol mol-1 PAR. Study revealed that irrespective of variety, -1 2 mean value of light saturation point (LSP) was 711 μmol mol , with mean net photosynthetic rate (Pn) of 8.8 μmol CO2 m s-1. Among the nine varieties, Amrapali and Tomy Atkins showed maximum LSP and highest Pn. of 13.28 and 11.53 μmol 2 -1 -1 CO2 m s , respectively, while cv. Kesar and Hybrid Ratna had lower LSP of 400 and 500 μmol mol , Pn. of 5.15 and 5.11, 2 -1 -1 μmol CO2 m s , respectively. Under low light intensity of 100 μmol mol , cv. Amrapali and Tomy Atkins showed maximum 2 -1 Pn. (3.51 and 2.82 μmol CO2 m s ) hence found more suitable for sustaining diffuse sunlight conditions during rainy season of Konkan.
PH378 (IPPC0451) Modulation of phosphoenolpyruvate carboxylase by abscisic acid and fusicoccin in the leaf discs of Amaranthus hypochondriacus Bindu Prasuna Aloor*, Uday Kiran Avasthi and A.S. Raghavendra Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad-500046, Andhra Pradesh, India *Presenting author: [email protected]
The kinetic and regulatory properties of phosphoenolpyruvate carboxylase (PEPC) are modulated by dark/light transitions and variation in temperature. Since the reports on hormonal modulation are limited, we examined the activity/regulation of PEPC by abscisic acid (ABA, a plant hormone) and fusicoccin (FC, an antagonist of ABA action) in the leaf discs of Amaranthus hypochondriacus. Incubation with 20 μM ABA for 1 h increased the activity of PEPC in leaf discs by > 40% in light and by 30% in dark. Butyric acid decreased PEPC activity and restricted the stimulation by ABA, while methylamine increased the PEPC activity and stimulated the effect of ABA. ABA enhanced the levels of PEPC-protein as well as its mRNA, both in dark and in light. Butyric acid/methylamine modulated the changes induced by ABA of PEPC-protein and mRNA levels, indicating that acidification/alkalization of leaf discs was very important. Since FC is known to antagonise/ reverse the effects of ABA, we have tested the effect of FC with or without ABA on PEPC properties. FC (10 μM) inhibited PEPC activity by 50% in dark as well as under illumination. The responses of PEPC to FC were subdued in the presence of 20 μM ABA. Significant increase in protein levels were observed in ABA treated leaf discs, while there was a decrease in PEPC protein levels in FC and butyric acid treated ones. We conclude that the enhancement by ABA of PEPC activity was due to alkalization and an increased protein levels.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 306 Session H : Photosynthesis
PH379 (IPPC0468) Variations in photosynthesis and light induced respiration in rice under different nitrogen treatments and in temperature gradient tunnel Rajesh Kondamudi*, Narasimha Swamy Konduri, Sailaja Bhogireddy, Veerendra Jaldhani, Suchandranath B Munnam, Satendra K Mangrauthia, Raghuveer Rao Puskur, Subrahmanyam Desiraju, Sarla Neelamraju and Sirapati Rao Voleti Indian Institute of Rice Research, Rajendranagar, Hyderabad-500030, Telangana, India *Presenting author: [email protected]
Rice crop was observed to be vulnerable to the climate change. Breeding for climate resilience in rice should include morpho-physiological characterization of existing germplasm. Five heat tolerant rice varieties were identified earlier (Somaly- 2-023-3-5-1-2-1, IR 55178, GQ-25, SG-26-120, IR-82310-B-B-67-2) along with a tolerant check Nagina 22 and a susceptible check Vandana were studied for seven consecutive seasons in four nitrogen treatments with elevated temperature (5±1°C above ambient) to understand the impact of elevated temperature and nitrogen fertilization. Leaf characteristics, morphological features, yield and yield attributes were recorded. Photosynthesis (A) and light induced respiration (LIR) were also studied using oxygraph. The A decreased with time in leaf samples, 14.57 μmoles m-2 s-1 (1d), 14.37 μmoles m-2 s-1 (2d) and 10.03 μmoles m-2 s-1 (3d) and A and total carbon were found to be in positive association. Conversely, the LIR varied greatly with 10.44 (1d), 13.11 (2d) and 9.60 μmoles min-1 s-1 (3d). The C/N ratio was decreased in leaf with time in almost all the treatments and varieties and increased with increased nitrogen doses but not under elevated temperature. Under heat stress, gene expression was also studied. Heat shock transcription factor (OsHsfA2a) cell wall integrity protein (CWIP) and sucrose phosphate synthase (SPS) genes showed increased expression. The expression of Osfd gene (iron-sulphur cluster binding-protein) showed a definite pattern between heat tolerant and susceptible germplasms. GQ-25 and Vandana showed down-regulation but other germplasm showed increased expression under elevated temperature. The interaction among N, P, K and Si elements and their role in making the rice crop resilient to high temperature was focussed.
PH380 (IPPC0494) Nitrogen assimilation is well coordinated with photosynthetic efficiency in pigeonpea grown under elevated CO2 Rachapudi Venkata Sreeharsha* and Attipalli Ramachandra Reddy Department of Plant Sciences, University of Hyderabad, Hyderabad- 500046, Andhra Pradesh, India *Presenting author: [email protected]
Pigeonpea (Cajanus cajan (L.) Millspaugh; Fabaceae) is an important food legume crop of equatorial and semi-arid parts of the world. A short duration pigeonpea cultivar (ICPL 15011) was grown under elevated CO2 (550 ppm) in open top chambers to understand the relation between photosynthetic physiology and N assimilation. Elevated CO2, pigeonpea grown showed significantly high net photosynthetic rates (A’) and foliar carbohydrate content (36% and 43%, respectively), which further manifested in dry biomass at harvest, with an increment of 29% over the ambient plants. Mass and number of nodules exhibited prodigious accretion in elevated CO2 grown plants, showing 58% increase in nodule mass ratio thereby overcoming the N limitation and resulting lack of photosynthetic acclimation. The high levels of N and amino acid contents in the leaf and root tissues of plants grown under elevated CO2 can be attributed to the increased levels of glutamine synthetase (GS) as evidenced by western blotting and qRT PCR studies. Further, GS and glutamate synthase
(GOGAT) showed significantly high activities in leaf and root tissues in elevated CO2 grown plants. To understand the transcriptional regulation of GS, the ORF sequence was characterized by cloning into E.coli BL21 cells, the expressed protein was purified and the sequence was confirmed by MALDI-TOF. In conclusion, this study revealed that lack of photosynthetic acclimation in pigeonpea under elevated CO2, can be attributed to the effective functioning of the N assimilatory enzymes including GS and GOGAT.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 307 Session H : Photosynthesis
PH381 (IPPC0510) Carbon sequestration and biomass yield in short rotation coppice mulberry (Morus spp.) grown under
CO2-enriched atmosphere K. Madhana Sekhar* and Attipalli R. Reddy Department of Plant Sciences, University of Hyderabad, Hyderabad-500046, India *Presenting author: [email protected]
Present study was conducted to elucidate the responses of mulberry under predicted 2050 levels of elevated CO2 -1 atmosphere (550 μmol mol ). Elevated CO2 grown mulberry plants showed significantincrease in light saturated photosynthetic rates (A’) throughout the experimental period by increasing intercellular CO2 concentration (Ci) despite reduced stomatal conductance (gs). Reduced gs was linked to decrease in transpiration (E) resulting in improved water use efficiency (WUEi). Increased in vivo maximum rate of RuBP carboxylation (Vcmax), in vivo maximum rate of electron transport driving RuBP regeneration (Jmax), light and CO2 saturated photosynthetic rates (Amax), apparent quantum efficiency
(AQE), photosynthetic nitrogen use efficiency (PNUE) and photosystem-II associated electron transport rate (ETRmax) clearly showed lack ofphotosynthetic acclimation in elevated CO2 grown mulberry. Changes in Vcmax and Jmax were positively correlated with foliar nitrogen (NA) levels. Significant rise in transcript levels associated with key regulatory enzymes of Calvin cycle, which are involved in both RuBP carboxylation and its regeneration,were believed to facilitate efficient carbon sequestration resulting in superior biomass yields (45%) in elevated CO2 grown plants compared to their controls.
Our data strongly suggest that short rotation (<1 year) with mulberry plantations should be effective to mitigate rising CO2 levels as well as for the production of carbon neutral renewable bio-energy.
PH382 (IPPC0514) Photothermal indices in relation to physiology, phenology and yield traits of rice genotypes under different sowing conditions V. Ravichandran1*, S. Jeyapriya2 and S. Robin1 1Department of Rice, 2Department of Crop Physiology, Tamil Nadu Agricultural University, Coimbatore-641003, India *Presenting author: [email protected]
Rice productivity is highly dependent upon prevailing weather parameters like temperature, day length and solar radiation are important natural resources which affect the rice productivity to a greater extent. Sowing time and selection of variety may play the deciding role in performance of rice production under changing climatic scenario. The proposed study was conducted to reveal the photothermal indices on physiology, phenology and yield components of ten rice genotypes under three sowing windows (June 9th, June 23rd and July 7th) for identifying the climate resilient rice genotypes suitable for wider climatic change. Among the sowing dates, the rice genotypes sown on June 9th reflected significantly higher plant height and total dry matter production. Physiological characteristics like gas exchange parameter and light transmission ratio was desirable in rice genotypes sown under June 9th and the long duration genotypes surpassed other genotypes for physiological characteristics, especially the genotypes CB-05-022 and IET 20924. Regarding phenology June 9th consumed more number of days to reach different growth stages in the rice genotype CB-05-022. The photothermal indices i.e. growing degree days, photothermal units, heliothermal units, heat use efficiency and radiation use efficiency were recorded maximum under June 9th sowing where, they took longer vegetative growth duration to produce higher grain yield by utilizing heat units and natural resources efficiently under favorable weather conditions. To conclude sowing date June 9th favoured better growth and yield in rice genotypes CB-05-022 which utilize heat units efficiently under favourable environment to provide better yield than other genotypes used.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 308 Session H : Photosynthesis
PH383 (IPPC0521) High light induced changes in organization, protein profile and function of photosynthetic machinery in Chlamydomonas reinhardtii Sai Kiran Madireddi, Elsin Raju Devadasu, Rajagopal Subramanyam and Srilatha Nama* Department of Plant Science, University of Hyderabad, Hyderabad-500046, Andhra Pradesh, India *Presenting author: [email protected]
The green alga Chlamydomonas (C.) reinhardtii is used as a model organism to understand the efficiency of photosynthesis along with the organization and protein profile of photosynthetic apparatus under one how exposure to various intensities of high light. Chlorophyll (Chl) a fluorescence induction, OJIPSMT transient decreased with increase in light intensity indicating the reduction in photochemical efficiency. Further, circular dichroism studies of isolated thylakoids from high light exposed cells showed considerable change in the pigment-pigment interactions and pigment-proteins interactions. Furthermore, the organization of supercomplexes from thylakoids was studied, in which, one of the hetero-trimer of light harvesting complex (LHC) II was affected significantly in comparison to other complexes of LHC’s monomers. Other supercomplexes, PSII reaction center dimer and PSI complexes was reduced. Additionally, immunoblot analysis of thylakoid proteins revealed that PSII core proteins D1 and D2 were significantly decreased during high light treatment. Similarly, the PSI core proteins PsaC, PsaD and PsaG were drastically changed. Further, the LHC antenna proteins of PSI and PSII were differentially affected. From our results it is clear that LHC’s are damaged significantly, consequently the excitation energy is not efficiently transferred to the reaction center. Thus, the photochemical energy transfer from PSII to PSI is reduced. The inference of the study deciphers the structural and functional changes driven by light influence; it may therefore provide plants/alga to regulate the light harvesting capacity in excess light conditions.
PH384 (IPPC0575) Photosynthesis and growth responses of mustard (Brassica juncea L. cv Pusa Bold) plants under free air carbon dioxide enrichment (FACE) condition Kamal Ruhil*, Sheeba, Altaf Ahmad, Muhammad Iqbal and B.C. Tripathy School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Increased atmospheric (CO2) is likely to affect photosynthesis, plant growth, and yield potential of plants. Mustard (Brassica juncea L.) is an important oil seed crop that is widely grown in India. Therefore, the impact of elevated (CO2) (585 μmol -1 mol ) on pigment and protein content, chlorophyll a fluorescence, photosynthetic electron transport reactions, CO2 assimilation, biomass production, and seed yield potential was measured in B. juncea cv. Pusa Bold grown inside free air carbon dioxide enrichment (FACE) rings installed on the campus of Jawaharlal Nehru University, New Delhi, India. Plants were grown for three consecutive winter seasons (2010-11, 2011-12 and 2012-2013) in ambient (385 μmol mol-1) or -1 elevated (585 μmol mol ) (CO2), in field conditions. Elevated (CO2) had no significant effect on the minimal chlorophyll fluorescence (F0), while the quantum efficiency of Photosystem II, measured as the ratio of variable fluorescence (Fv=Fm– F0) to maximum fluoresence (Fm) (Fv/Fm), increased by 3%. Electron transport rate of photosystem I and photosystem II, and the whole chain electron transport rates increased by 8% under elevated (CO ). However, the net photosynthesis ≈ 2 rate increased by 50% in three growing seasons under elevated (CO2) condition. The stomatal conductance and transpiration rate decreased resulting in higher photosynthetic water use efficiency. The photosynthesizing surface, i.e., leaf area index substantially increased leading to higher biomass and seed yield under elevated (CO2) condition. Acclimatory down regulation of photosynthesis and plant productivity was not observed in three consecutive growing years, suggesting that in the absence of nutrient limitation, B. juncea is highly responsive to elevated CO2, whose yield potential shall increase in changing climatic conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 309 Session H : Photosynthesis
PH385 (IPPC0610) Size effect of zinc oxide nanoparticles on photosynthesis and oxidant response in Oryza sativa Prabhat Sharma and Maria Vera Da Costa* Department of Botany, Goa University, Taleigao Plateau, Panaji-403206, Goa, India *Presenting author: [email protected]
Inevitable release of nanoparticles (NPs) in the environment due to waste generated by nanodevices or during nanomaterial manufacturing process is a major concern. The physiological behaviour of Oryza sativa var. Jyoti treated with zinc oxide nanoparticles (ZnO NPs) of 50 and 100 nm size were hydroponically studied. Both sizes of ZnO NPs showed negligible effect on germination. Uptake of NPs was accompanied by reduction in shoot, root length, and shoot biomass, also, number and size of stomata decreased and number of trichomes increased. In contrast to non-metallo nanoparticles, no deposition of ZnO NPs in the intercellular spaces of root parenchyma was seen, however, a decrease in the thylakoid number per granum of chloroplast at 1000 mg L-1 ZnO (100 nm) was observed. The maximum quantum efficiency of PSII (Fv/Fm ratio), photochemical quenching (qP), photosynthetic rate (PN), stomatal conductance (gs), transpiration rate (E) and photosynthetic pigment content were more significantly affected by 50 nm compared to 100 nm ZnO NPs. Proline and ascorbate content increased while lipid peroxidation showed no significant increase on exposure with both sizes of ZnO NPs. Dissolution of ZnO NPs into metal ions was found to be far below the toxicity threshold attributing the phytotoxic effect observed to NPs per se. Our results demonstrate that 50 nm sized NPs were more toxic compared to 100 nm sized ZnO NPs on the photosynthetic processes with significant increase in osmotic stress without causing lipid peroxidation.
PH386 (IPPC0626) Inhibitory effects of metallo nanoparticle on photosynthesis are due to reduction in thylakoid membrane in the grana region of chloroplasts rather than oxidative damage to proteins and lipids Prabhat Kumar Sharma* Department of Botany, University of Goa, Goa, India *Presenting author: [email protected]
The physiological and biochemical behaviour of rice (Oryza sativa var. Jyoti) treated with CuO and ZnO nanoparticles (NPs) in the range of 0-1000 mg L-1 was studied hydroponically. The treatment resulted in accumulation of the NPs, many fold higher in roots than shoots indicating exclusion of metallo nanoparticle at root level. The accumulation was further observed inside cells, especially in the chloroplasts and caused decreased stacking and structural deformation of thylakoids per granum. Photosynthetic rate, transpiration rate, stomatal conductance, maximal quantum yield of PSII, biomass and photosynthetic pigment contents decreased, with a complete loss of PSII photochemical quenching at 1000 mg L-1 of NPs treatment. Significant level of osmotic stress was evident by increased proline content as a result of the treatment, however, little or no oxidation to protein and membrane lipids, studied as protein carbonyl and MDA content respectively, was observed. Elevated expression levels of ascorbate peroxidase and superoxide dismutase but no oxidative damage to protein and membrane lipids suggest that production of ROS due to the treatment was within the inherent capacity of its metabolism. Results showed that dissolution level of NPs into ions was much below the threshold of ion toxicity, indicating toxic effect due to the NPs rather than their ions. Results demonstrated that NPs caused inhibitory effect mainly due to the decreased stacking of thylakoid membrane in the grana region of chloroplasts leading to smaller LHC II thus lower production of ROS, and not due to the oxidative damage as seen with bulk particles.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 310 Session H : Photosynthesis
PH387 (IPPC1005) Antioxidant system-mediated regulation of chlorophyll biosynthesis pathway in vascular plant Cucumis sativus upon exposure to methyl viologen and high light stress Aarti P. Dhepe* and Komal A. Joshi Institute of Bioinformatics and Biotechnology, S.P. Pune University, Pune-411007, India *Presenting author: [email protected] Reactive Oxygen Species (ROS), being highly reactive in nature can cause oxidative damage to various biological molecules. However, they can also act as a signalling molecule in diverse cellular processes. So, their generation and accumulation has been found to be very crucial for plant growth, development as well as defense. This outstanding potential of ROS is due to the development of extremely competent antioxidant systems, which either by over expression or depletion maintain their appropriate concentration in plants. In this study, a regulatory relation between antioxidant system and chlorophyll biosynthesis pathway has been studied. Cucumber cotyledons, etiolated (E) and green (G, 14h illuminated), were treated with methyl viologen (MV) or high light (HL, 400-500 uE m-2 s-1) and the effects were studied on chlorophyll biosynthesis pathway as well as on antioxidant system. ROS generation was confirmed by proline and H2O2 concentration measurement. It was observed that chlorophyll biosynthesis was affected at the level of ALA synthesis and protoporphyrinogen IX. Measurement of MDA contents showed that inhibition observed was not due to loss of membrane integrity. When the activities of several antioxidant enzymes were assayed, it showed that ascorbate peroxidase activity was reduced in E and G cotyledons upon MV treatment and in HL-treated E cotyledons. Catalse activity was also found to be severely impaired in HL-exposed E cotyledons. Ascorabate and lutein contents were found to be decreased significantly in MV treated E and G cotyledons and E-HL cotyledons. This suggested that significant decrease in the activities of H2O2–scavenging antioxidants might have resulted in increased concentration of H2O2 which in turn might have caused inhibition of chlorophyll biosynthesis pathway.
PH388 (IPPC1038) Identification and characterization of sugarcane genotypes for photosynthetic efficiency, moisture stress tolerance and high yield T. Sujatha*, V.S.P. Lalitha and K. Jhansi Crop Physiology, Sugarcane Research Station, Vuyyuru-521165, India *Presenting author: [email protected] Six early pre-release sugarcane clones were tested against standard Co 6907 for their suitability under moisture stress conditions at Sugarcane Research Station, Vuyyuru during 2013-14 and 2014-15 seasons. Only two irrigations were given at 10th and 40th day after planting in addition to at the time of planting. Among the clones tested 2007 V 127 (97.77 t/ha) and 2007 V 131 (96.13 t/ha) recorded significantly higher cane yield when compared to standard Co 6907 (80.84 t/ ha) where as the clone 2007 V 129 (86.60 t/ha) was on par with the standard. Johari etal 1998 also reported that growth and yield affected due to soil moisture tension but the magnitude varied with the clones. Length of the millable cane was higher in 2007 V 131 and 2007 V 129. Higher diameter of the cane was recorded in 2007 V 131. The clones 2007 V 127 (79.55%) and 2007 V 131 (79.53%) maintained more leaf sheath moisture percent, the same clones 2007 V 127 (79.20) and 2007 V 131 (79.07) were also recorded more relative water content percent during formative stage which coincided with summer. Dead canes recorded in those clones were also less compared to other cones. Higher percent juice sucrose was recorded in 2007 V 127(21.39 %) followed by 2007 V 131 (21.35 %) and 2007 V 129 (21.04 %). The enzyme superoxide dismutase was significantly higher in 2007 V 131 (1.17) followed by 2007 V 121 (1.14) and 2007 V 127 (1.08) which shows the tolerance of these clones to moisture stress. The clones 2007 V 127 and 2007 V 131 with higher RWC % leaf sheath moisture %, juice sucrose % length of the millable cane and cane yield were found tolerant to moisture stress conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 311 Session I : Genomics-assisted Plant Improvement
PI389 (IPPC0023) Quantitative proteomic analysis for elucidation of mechanism governing salt stress tolerance in germinated seedlings of pigeonpea Vandna Rai* National Research Centre on Plant Biotechnology, New Delhi-110012, India *Presenting author: [email protected]
Pigeonpea (Cajanus cajan), a major source of dietary protein, is often subjected to salinity stress under different agroclimatic conditions thereby severely limiting its productivity. Proteomic analysis is a viable approach for identifying set of proteins that may have potential roles in conferring tolerance to salinity and/or other abiotic stresses. In this study, the effect of varying concentrations of NaCl treatment for 7 d were evaluated on the developmental responses of root system architecture (RSA) and shoots of pigeonpea. Developmental responses of these traits were significantly affected during growth at 150 mM NaCl. A label-free comparative Q TOF, LC-MS/MS technique was employed for qualitative and quantitative proteomics analysis of roots and shoot grown under saline-free (0 mM NaCl) and saline-stress (150 mM) conditions. A total of 119 differentially expressed (DE) proteins were identified of which 78% of them were represented in both roots and shoots. Interestingly, some of them such as Ras related and RNA-dependent RNA polymerase were detected only in the roots of the seedling that were subjected to salinity stress. In addition, a few proteins i.e., Ribulose-1,5-bisphosphate carboxylase/ oxygenase large subunit and Dof 34 were detected in saline-stress shoot. The analysis suggested that these proteins in roots and shoots may have a role in salinity stress response. Further, protein interaction network (PIN) was developed using the String software with Confidence Scores >0.7 for identifying key regulators of salt stress tolerance in pigeonpea.
PI390 (IPPC0187) Dynamics of long non-coding RNAs responsive to Sclerotinia sclerotiorum infection in canola (Brassica napus) Raj Kumar Joshi1*, Swati Megha2, Urmila Basu2 and Nataraj Kav2 1Centre of Biotechnology, Siksha O Anusandhan University, Bhubaneswar-751003, Odisha, India 2Department of Agricultural Food and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada *Presenting author: [email protected]
The biggest constraint for canola (Brassica napus) production worldwide is the Sclerotinia stem rot caused by necrotrophic, non host specific phytopathogen Sclerotinia sclerotiorum. Short non coding RNAs have been proven to play important roles in the regulation of stress responses in canola but the functions of long non coding RNAs (lncRNAs) largely remains unknown. In the present study, we used a strand specific RNA sequencing approach to identify lncRNAs responsive to S. sclerotiorum infection in B. napus. A large set of B. napus RNA-seq data were examined with high sequencing depth under control and inoculated conditions and 3181 lncRNA candidates were identified. A total of 931 lncRNAs were found to be pathogen responsive, 12 of which were confirmed by RT-qPCR. Additionally, RT-qPCR verification of a set of sense/antisense transcript pairs revealed contrasting expression patterns suggesting steric clashes of transcriptional machinery. Forty one lncRNAs served as precursors for several stress responsive microRNAs having significant role in the regulation of pathogen defense. B. napus lncRNA, TCONS_00000966 antisense to plant defensin gene PDF1.2 was significantly induced suggesting its involvement in the transcriptional regulation of defense responsive genes during S. sclerotiorum infection. Our results suggested that, RNA-Seq provide a comprehensive view of long non coding transcripts and lncRNAs are important constituent of the antifungal network in B. napus.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 312 Session I : Genomics-assisted Plant Improvement
PI391 (IPPC0220) LeasyScan - a novel semi-field platform to phenotype traits controlling plant water budget Grégoire Hummel1*, Vincent Vadez2 and Jana Kholova2 1Phenospex B.V., Jan Campertstraat 11, 6416 SG Heerlen, Germany 2International Crops Research Institute for Semi-Aird Tropics (ICRISAT), Patancheru-502324, Telangana, India Presenting author: [email protected]
We describe the concept and realisation of a high-throughput phenotyping platform (LeasyScan) combined with lysimetric capacity, to assess canopy traits affecting water use. The platform is based on a novel 3D scanning technique to capture canopy development, a scanner-to-plant concept to increase throughput (2500 plants/h), and analytical scales to combine gravimetric transpiration measurements. We present how the technology functions, how data are visualised via a web- based interface, and how data extraction and analysis is interfaced through ‘R’ libraries. Close agreement between scanned and observed leaf area data of individual plants in different crops was found (R2 between 0.86 and 0.94). Similar agreement was found when comparing scanned and observed area of plants cultivated at densities reflecting the field conditions. Example of the monitoring, the plant transpiration by the analytical scales is presented. Moreover, we present some on-going applications of the platform to target key phenotypes: (i) the comparison of the leaf area development pattern of fine mapping recombinants of pearl millet; (ii) the leaf area development pattern of pearl millet breeding material targeted to different agro-ecological zones; (iii) the assessment of the transpiration response to high VPD in sorghum and pearl millet.
PI392 (IPPC0227) Genome-wide DNA polymorphisms in rice genotypes with contrasting behaviour under low Phosphate Poonam Mehra*, Bipin Pandey and Jitender Giri National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Soil phosphorus (P) deficiency is one of the major challenges to the rice cultivation world-wide. Modern rice genotypes are highly P-responsive and rely on high input of P fertilizers. However, low P tolerant traditional cultivars and landraces have genetic potential to sustain well under low P conditions. Identification of high resolution DNA polymorphisms (SNPs and InDels) in such contrasting genotypes can serve as good resource for marker assisted breeding for improvement of low P sensitive high yielding cultivars. Therefore, high quality DNA markers in low P sensitive modern genotype, PB1 and tolerant traditional genotype, Dular were identified by whole genome resequencing. Approximately, 2.3 million and 2.9 million high quality polymorphisms with an average read depth of >24X were identified in PB1 and Dular, respectively with reference to Nipponbare genome. Several non-synonymous and regulatory DNA polymorphisms having potential functional significance were mapped to key Phosphate Starvation Responsive (PSR) and root architecture genes/QTLs in rice. To deduce more promising SNPs/InDels potentially associated with low P tolerance, all Dular and PB1-specific variants underlying PSR genes were compared with variants in another low P tolerant rice genotype, Kasalath. Further, all polymorphisms underlying PSR genes were classified on the basis of their presence in low P sensitive genotypes (PB1 and Nipponbare) or low P tolerant genotypes (Dular and Kasalath). These identified variants can serve as a good source of genetic variability for improving low P tolerance and root architecture of high yielding modern genotypes.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 313 Session I : Genomics-assisted Plant Improvement
PI393 (IPPC0237) Ectopic expression of Pennisetum glaucum AsA-GSH pathway in ripened tomato fruits for enhancing postharvest shelf life by reduction in oxidative damage Krishnamurthy Sathelly*, Tanushri Kaul and Murtaza Malik Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-67, India *Presenting author: [email protected]
Globalization has led to an increasing demand for quality improvement and the wider adoption of quality standards for fruits and vegetables. Plant transformation and regulated gene expression offers an integrated approach to tackle complex traits such as post-ripening oxidative deterioration of fruits. Genes that code for enzymes involved in key oxidative stress detoxification via the ascorbate-glutathione pathway reveal a decline in their activities and also non-enzymatic antioxidants ascorbate and glutathione content decreases during post ripening of tomato fruits. Our strategy to deal with postharvest oxidative damage in tomatoes involves the overexpression of ascorbate-glutathione pathway genes from a stress adapted
C4 plant – Pennisetum glaucum driven by different fruit ripening-specific promoters for scavenging the excess ROS generated in tomato fruits during postharvest storage through agro-mediated transformation. Stable integration of transgenes was confirmed by PCR and Southern analyses in T0 transgenic plants. Northern analysis in red-ripe stage of T1 fruits revealed elevated transcript levels of transgenes in comparison to transgenic mature green and WT fruits. Activities of AsA-GSH pathway enzymes significantly increased (80-100%) in red-ripe tomato fruits in comparison to WT fruits. Postharvest decay symptoms appeared within 7 ± 0.8 days in WT fruits, whereas these developed after 31.8 ± 0.5 days in transgenic red-ripe fruits that indicated delayed postharvest senescence. Results suggested an upregulation of AsA-GSH cycle in red-ripe fruits during postharvest stress conditions. Fruit-specific enhanced expression of antioxidant enzymes from AsA-GSH pathway in tomato fruits effectively scavenged terminal ROS and minimized postharvest deterioration, in turn enhancing their shelf life.
PI394 (IPPC0243) Protein-Protein Interaction and gene co-expression network of rice TCPs Vasudha Bhardwaj1*, Aleena Francis2, Ananda Rao Saketi3, Abhishek Kumar Anuj1, Suchismita Roy3, Manoj K. Sharma1, Ashwani Pareek3 and Rita Sharma2 1School of Biotechnology, 2School of Computational and Integrative Sciences, 3School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
TCP is a plant-specific family of transcription factors with its name derived from the founding members, Teosinte branched 1 (TB1) of maize, Cycloidea (CYC) of Antirrhinum and Proliferating cell factors (PCFs) of rice. Initially, identified as key players in regulating plant growth and development, members of TCP family have recently emerged as multifaceted players engaged in abiotic stress tolerance as well as plant immunity. They can modulate wide range of biological processes by regulating variety of hormonal pathways and signaling cascades. Rice genome encodes 23 TCP proteins divided into two classes. Class I factors activate plant growth; whereas, class II factors have been shown to negatively regulate growth-associated genes. The ability of class I and II factors to antagonize each other reflect a complex mechanism of gene regulation. However, genetic redundancy is a major hindrance in elucidating the precise mechanism of their action. Also, TCP proteins are known to form homo and heterodimers for their function but how the choice of interacting partners dictate the transcriptional outcome remains unresolved. In this study, we have identified rice interologs derived from Arabidopsis TCP interactome and validated the pairwise-interactions using yeast two-hybrid assay. Further, to dissect the functionally redundant gene pairs, we have performed a comprehensive expression and coexpression analysis of TCP genes in rice. The experimentally verified interactome coupled with sequence analysis, interologs, transcriptional profiling and coexpression analysis would be highly useful to elucidate redundancy and gene functions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 314 Session I : Genomics-assisted Plant Improvement
PI395 (IPPC0255) Genomic assisted breeding for introgression of gynoecious trait in Indian monoecious cucumber (Cucumis sativus) cv. Pusa Uday Tusarkanti Behera, Harshawardhan Choudhary*, Anilabh Dasmunshi, Rajni Karn and Brihama Dev Indian Agricultural Research Institute New Delhi-110012, India *Presenting author: [email protected]
Indian cucumber cultivars exhibit monoecious sex expression coupled with crown fruit inhibition which is considered as one of the reasons for lower productivity. Gynoecism has been extensively exploited in heterosis breeding for enhancing productivity of cucumber worldwide but exotic gynoecious lines are not stable at higher temperature and they possess very few lateral branches which are not suitable for open field conditions. The introgression of gynoecious trait with marker assisted breeding can improve the productivity of Indian cucumber. The F2 population from cross G 421 (exotic gynoecious) x Pusa Uday (Indian monoecious) was used for mapping, with 6 markers from the genomic region of F locus. The markers SSR-13251 and SSR- 15516 were found to be linked to F locus at 1.5 and 4.5 cM respectively. One gene based SCAR marker clearly distinguished gynoecious and monoecious plants in all F2 population. These 3 markers were used in foreground selection from 192 BC1F1 individuals and 18 heterozygous plants for F locus phenotypically similar to Pusa Uday were selected. These plants were selfed to produce the BC1F2 and backcrossed with Pusa Uday to develop BC2F1 population. The genotyping of these two populations was performed with these 3 markers and phenotypic selection was made for the traits similar to Pusa Uday with gynoecism. Genotypes with homozygous for F locus, stable phenotypic expression of gynoecious trait and growth and fruit characters matching to Pusa Uday were selected. These plants will be further evaluated for horticultural traits in different environment for varietal development.
PI396 (IPPC0264) Engineering rice with PgOGG1, a bifunctional DNA glycosylase/AP lyase enhances abiotic stress tolerance and seed longevity V. Mohan Achary*, James Donald, Dhirendra Fartyal, Mrinalini Manna, Vijay Reddy, Ravi Kumar, Bhabesh Borphukan and M.K. Reddy Plant Molecular Biology Group, International Centre for Genetic Engineering & Biotech., Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Reactive oxygen species (ROS) are the by-products of normal cell metabolism in plants. However, under stress conditions including seed desiccation, storage and germination, ROS homeostasis is disturbed resulting in seed deterioration and therefore, decreased seed longevity. Notably damage to the genome induced by ROS has been recognized as an important cause of seed deterioration. The prominent DNA lesion induced by ROS is 7,8-dihydro-8-oxoguanine (8-oxo-G), that form base pair with adenine instead of cytosine during DNA replication and leads to GC/TA transversion. This altered base is removed by 8-oxoguanine DNA glycosylase and repaired by the subsequent BER enzymes. In this report, we identified and characterized OGG1 from the C4 plant Pennisetum glaucum (PgOGG1) that encodes 45.3 kDa protein showing significant sequence identity with plant orthologues as well as yeast and human OGG1 protein. The transcript of PgOGG1 was strongly up-regulated during seed desiccation and imbibition. Purified PgOGG1 specifically cleaves 8-OxoG:C mispair DNA duplex which is a characteristic of all bifunctional DNA-glycosylases/lyases. PgOgg1 was found to localize both in nucleus and mitochondria. Expression of PgOGG1 in rice showed reduced level of 8-oxo-dG and enhanced seed resistance to controlled deterioration treatment indicating a DNA damage-repair function of PgOGG1 in vivo. Furthermore, transgenic seeds exhibited increased germination ability under abiotic stresses induced by MV, NaCl and high-temperatures. Taken together, our results demonstrate that ectopic expression of PgOGG1 enhances seed longevity and abiotic stress tolerance in rice mediated by repair of the damaged DNA through elimination of 8-oxoG from the genome.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 315 Session I : Genomics-assisted Plant Improvement
PI397 (IPPC0285) Genome wide study of two component system and identification of key members of cytokinin signaling during nodule development in chickpea Manish Tiwari* and Sabhyata Bhatia National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Chickpea is the world’s second most grown legume, with Indian subcontinent being the principal chickpea producing and consuming region. Nitrogen fixation by symbiotic association increases its importance. Symbiotic association is in form of root nodule, which harbors bacteria and facilitates nitrogen fixation. Several studies have been carried out on the root nodule formation; and it has been clearly observed that cytokinin has very important role in nodulation. The expression of trans – zeatin gene in nodulation-deficient phenotype of Rhizobium mutant initiates nodulation. Cytokinin application also induced the early nodulin gene and formation of false nodules. Cytokinin signaling is achieved by two component system (TCS) in plants which is composed of sensor histidine kinases (HKs), histidine phosphotransferases (HPs), and response regulators (RRs) and it would be interesting to investigate their roles in cytokinin mediated nodulation in chickpea. Towards this, an exhaustive genome wide search of the chickpea genome for TCS members was conducted. Analysis of their conserved domains, and phylogenetic relationships of these protein-coding genes was carried out. The transcription levels of TCS genes in various organs and different nodule developmental stages along with environmental stresses were investigated. Our systematic analyses provide insights into the characterization of the TCS genes in chickpea, their involvement in root nodule formation and basis for further functional studies of such genes.
PI398 (IPPC0415) Phenotypic/genotypic evaluation of, and conservation strategy for, natural populations of endangered Boswellia Serrata Roxb. Shashank Mahesh1, Pramod Kumar1, Vivek Vaishnaw1, Naseer Mohammad1, Ashwani Kumar2 and Shamim Ansari3* 1Tropical Forest Research Institute, Jabalpur-482021, Madhya Pradesh, India 2Indian Council of Forestry Research and Education, Dehra Dun-248006, India 3Institute of Forest Productivity, Ranchi-835303, Jharkhand, India *Presenting author: [email protected]
Boswellia serrata Roxb. (salai) is an important species of dry deciduous forest and has been over exploited for its oleo gum-resin and excellent paper pulp in the central India. Consequently, there exists fragmentation of populations and poor natural regeneration. We investigated variability and status of populations in terms of wood fiber length and tree girth by sampling 20 trees each of 12 natural Madhya Pradesh populations of B. serrata existing in different agro-climatic conditions. The wood fiber length and tree girth exhibited 59.7% and 505.2% variations, respectively, across populations. There was small positive and negative kurtosis respectively for both parameters, which indicates uneven loss of genotypes or genetic drift. Khandwa population with the longest wood fiber length was 18.2% superior over Jabalpur population with the shortest wood fiber length. Tree girth of Chhindwara population was maximum and 109.2% more than that of Damoh population. The broad sense heritability values for wood fiber length (0.201) and tree girth (0.334), indicate that both traits are associated with fitness in natural populations. Further, QST estimates for both parameters exceeding FST estimates obtained with neutral molecular markers (RAPD and ISSR), indicate towards operation of divergent natural selection. Nevertheless, the findings encourage for transfer of diverse genotypes from one population to the other for facilitating out crossing within populations as well as production of viable seeds, circumventing inbreeding depression and genetic drift. The proposed strategy would promote natural regeneration and in situ germplasm conservation/enrichment, which would help rescue the species from endangered status.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 316 Session I : Genomics-assisted Plant Improvement
PI499 (IPPC0457) Transcription factor gene families from Papaver somniferum: Identification of regulatory factors involved in benzylisoquinoline alkaloid biosynthesis Parul Agarwal1*, Sumya Pathak2, Deepika Lakhwani1, Parul Gupta1, Mehar Asif1 and Prabodh Trivedi1 1Molecular Biology and Genetic Engineering, CSIR-National Botanical Research Institute, Lucknow-226001, India 2University of Lucknow, Lucknow-226007, India *Presenting author: [email protected]
Opium poppy (Papaver somniferum L.) is known for the biosynthesis of structurally diverse and pharmaceutically valuable benzylisoquinoline alkaloids (BIAs) which includes narcotic analgesic morphine, the cough suppressant codeine, the muscle relaxant papaverine and the anti-microbial agent sanguinarine and berberine. Coordinated transcriptional regulation of biosynthetic genes has emerged as a major mechanism controlling the production of secondary metabolites in plant cells. Transcription factors, one of the prime regulators of secondary plant product biosynthesis, might be involved in controlled biosynthesis of BIAs. In this study, identification of members of different transcription factor gene families using transcriptome datasets of 10 cultivars of P. somniferum with distinct chemoprofile has been carried out. WRKY was identified as the most represented transcription factor gene family in all the poppy cultivars. Comparative transcriptome analysis revealed differential expression pattern of the members of a set of transcription factor gene families among 10 cultivars. Utilizing the integrated transcript and metabolite profiling strategy, two members of WRKY and one member of C3H gene family were identified as potential regulators of the papaverine and thebaine biosynthesis, respectively. The phylogenetic analysis suggest their close relationship with members of these gene families from other plants species. The regulatory role of these transcription factors and their involvement in specific alkaloids biosynthesis is being investigated using virus induced gene silencing.
PI400 (IPPC0459) Identification and expression analysis of genes involved in biosynthesis of Azadirachtin and related triterpenoids in Azadirachta indica (Neem) Sweta Bhambhani1*, Deepika Lakhwani1, Ashutosh Pandey2, Mehar Asif1 and Prabodh Trivedi1 1Molecular Biology and Genetic Engineering, CSIR-National Botanical Research Institute, Lucknow-226001, India 2National Agri-Food Biotechnology Institute, Mohali, Punjab, India *Presenting author: [email protected] Azadirachta indica, commonly known as ‘Neem’ contains tetranoterpenoids synthesized by isoprenoid pathway, which play various functional roles in plant defense. The major bioactive components include Azadirachtin, Nimbin and Salannin, which serve as ecofriendly biopesticides and insecticides due to their various important roles as insect feeding deterrants, toxicants, disruptants of growth and development against a variety of insects, pests and nematodes. Numerous studies have been carried out in relation to phytochemistry and biomedical activity of the plant but limited information is available on genes involved in the biosynthetic pathway of azadirachtin and related triterpenoids. In the present study, we have identified genes involved in triterpenoid backbone and analyzed their expression in various plant parts of A. indica. Structural analysis of genes suggests significant conserved structure and high sequence homology with homologues genes from other plants. Expression analysis of genes in various plant parts suggests a weak correlation with phytochemical accumulation suggesting involvement of enzymes responsible for secondary transformation. As putative biosynthetic pathway of Azadirachtin suggests involvement of cytochrome P450 (CYP450) enzymes in secondary transformation, we identified putative CYP450s having significant correlation between Azadirachtin content and its expression. This is the first study to identify and correlate expression of putative genes involved in biosynthetic pathway of Azadirachtin biosynthesis from neem.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 317 Session I : Genomics-assisted Plant Improvement
PI401 (IPPC0467) Characterisation of Oxidosqualene Cyclase encoding genes involved in withanolide biosynthesis from Withania somnifera Aditya Vikram Agarwal1*, Parul Gupta1, Rahul Michael1, Deepak Chandra2 and Prabodh Kumar Trivedi1 1CSIR-National Botanical Research Institute, Lucknow-226001; 2Department of Biochemistry, Lucknow University, Lucknow-226007 *Presenting author: [email protected] Tri-terpenes, a diverse group of plant secondary metabolites, possess variety of biological activities related to the pharmaceutical, cosmetic and food sectors. Withania somnifera (L.) Dunal, a medicinal shrub has various medicinal properties as anti-arthritic, anti-aging, anticancer and physiological and metabolic restoration. These pharmacological activities have been correlated with the withanolides, C28-steroidal lactones derived from tri-terpenoids. Biosynthesis of withanolides involves one or both of committed upstream routes of isoprenogenesis-the cytosolic mevalonate (MVA) pathway and plastid localized 2-C-methyl-D-erythritol- 4-phosphate (MEP) pathway. These pathways direct the flux of the isoprene (C5) units into the tri-terpenoid pathway via a common C30-intermediate 2,3-oxidosqualene. In Withania, further biosynthesis from this C30 intermediate is partitioned by cycloartenol synthase (CAS) and an array of other oxidosqualene cyclases (OSCs), which together form a myriad of diverse triterpenoids. CAS leads to the formation of cycloartenol, a tetracyclic tri-terpene, which acts as a precursor for phytosterols and apparently withanolides. OSCs are positioned at a key metabolic sub-dividing junction executing an indispensable step in varied tri-terpenoid biosynthesis thus presenting favourable gene targets for redirecting metabolic flux towards specific secondary metabolites. However, involvement of specific OSCs towards biosynthesis of withanolides has not been elucidated yet in planta. In this study, involvement of genes encoding OSCs in withanolide biosynthesis has been demonstrated through virus-induced gene silencing (VIGS) approach. Expression analysis and phytochemical analysis suggest decreased level of transcripts for specific OSCs in VIGS silenced lines as well as their involvement in directing flux towards withanolide biosynthesis.
PI402 (IPPC0497) Mobilizing landrace variations for next generation wheat improvement Prashant Vikram1*, Juan Burgueno Ferrera1, Jorge Franco2, Carolina Saint-pierre1, Carolina Paola Sansaloni1, Amita Mohan3, Mathew Reynolds1, Carlos Guzman1, Pawan Kumar Singh1, Peter Wenzl1, Thomas Payne1, Kulvinder Singh Gill3, José Crossa1 and Sukhwinder Singh1 1International Maize and Wheat Improvement Center (CIMMYT), Mexico; 2Departamento de Biometría, Estadística y Computación, Facultad de Agronom&iacut; 3Washington State University, Washington, USA *Presenting author: [email protected] Slow yield potential gain, climate change effects, expected increase in future food demand and narrowed breeding germplasm pool has emphasized an increased use of gene bank resources. Food and agriculture organization (FAO) has accentuated on enhanced use of on-farm and gene bank diversity. Semi-dwarfing wheat varieties revolutionized grain production; however, these varieties could not deliver significant impact in rainfed areas. Preliminary evaluation of pre- breeding populations derived from the most popular wheat variety, PBW343 revealed the importance of semi-dwarfing genes for rainfed environments and suggested for enhanced infusion of useful landrace variations to the breeding germplasm pool. Unique efforts have been made in ‘Seeds of discovery’ project at International center for maize and wheat improvement (CIMMYT) to perform physiological, genetic and molecular characterization of gene bank resources focusing mainly on landraces. Genotyping-by-sequencing of 11000 wheat landraces (from Mexico and Iran) was carried out along with trait characterization for heat, drought and grain quality. Landrace core set was formulated following a unique strategy which involved a data matrix representing phenotypic and genotypic variables simultaneously. Genomic associations and novel alleles for agronomically important traits have been identified. Trait donors were identified for heat; drought, diseases, quality and yield. These resources are being strategically deployed through pre-breeding to achieve short term, medium term and long term goals. Landrace and other resources are being leveraged with NARS in South Asia, Africa and USA for multiple location evaluation. We welcome global wheat community to join hands and deliver high impact in respective environments.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 318 Session I : Genomics-assisted Plant Improvement
PI403 (IPPC0513) Unravelling the role of miR159-MYB65 module for regulating reproductive organ development in Brassica Saurabh Anand*, Neer Singh and Sandip Das Department of Botany, University of Delhi, Delhi-110007, India *Presenting author: [email protected]
Reproductive development in plants is an outcome of complex interplay of genes and genetic networks. Unravelling the components of this network and understanding these interactions has helped in gaining knowledge about the genes and small RNAs involved in androecium and gynoecium development. MicroRNA159 that targets two GA-MYB transcription factors MYB33 and MYB65, were shown to affect reproductive development in Arabidopsis thaliana. To understand their precise role, miR159 and MYB65 have been identified from Brassica rapa and B. juncea utilizing comparative genomics approaches, and cloned. Transgenic lines over-expressing miR159 in A. thaliana Col-0 genetic background exhibit defects in flowering time, anther dehiscence, anther and pistil morphology, pollen-pistil interaction, pollen viability and fertility, and seed set. We have performed transcriptome analysis including RNA-sequencing and semi-qPCR of the transgenic and the WT lines to gain an insight into the global changes and perturbation accompanying developmental defects. Additionally, the role of hormones in reproductive organ development, especially GA, is being studied through experiments involving external applications. Experiments are also being performed using Brassica juncea owing to its importance as oil-yielding crop. Towards this goal, transgenic plants have been generated and are currently being analysed.
PI404 (IPPC0516) Analysis of root system architecture in chickpea Vikas Dwivedi*, Swarup Kumar Parida and Debasis Chattopadhyay National Institute of Plant Genome Research, Aruna Asif Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Chickpea (Cicer arietinum) is the third most consumed legume crop in the world. It is a major source of protein and, therefore, a very important food crop in developing countries. Due to several biotic and abiotic factors, annual crop yield is severely affected all over the world. To develop agricultural sustainability, chickpea is an important legume crop having excellent ability to fix atmospheric nitrogen biologically. A correlation between root system architecture and resistance to water stress has been found in several crop plants and breeding attempts have focused on obtaining cultivars with larger root systems. Development of root biomass and its architecture helps the plant to access water more efficiently from soil. The manipulation of root architecture could minimize the negative impact of a number of abiotic factors and can improve yield. Root System Architecture (RSA) is different between two species and also within the species. RSA is a highly plastic trait. Many methods are used for study of the root system. The development and architecture of roots hold potential for the use and manipulation of root characteristics to both increase yield and optimize agricultural land use. To understand the variation in chickpea root system architecture, we have studied about 41 different varieties of chickpea. Root system architecture (RSA) was analyzed for these varieties using different tools. We have also studied the root architecture of one mapping population (JGK3 X Himchana 1). Our main objective was concentrating on identification of genetic loci for root development in two chickpea genotypes with contrasting root traits.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 319 Session I : Genomics-assisted Plant Improvement
PI405 (IPPC0554) Functional characterization of MYC2 in disease defense Janesh Kumar Gautam1*, Mrunmay Giri2, Sudip Chattopadhyay3 and Ashis Kumar Nandi1 1 School of Life Sciences, Jawaharlal Nehru University, New Delhi-110 067, India 2University of North Texas, USA 3National Institute of Technology, Durgapur-713209, West Bengal, India *Presenting author: [email protected]
Plants have evolved a complex immune system that is tightly regulated at transcriptional level by myriad of transcription factors. MYC2, a basic helix loop helix (bHLH) transcription factor plays important roles in growth, development and disease defense. MYC2 negatively regulates blue light mediated hypocotyl elongation. Reports suggest that MYC2 also acts as a negative regulator of basal defense against bacterial pathogens. Our results show that MYC2 OEX plants have very strong level of resistance than myc2 mutants against Pseudomonas syringae. MYC2 binds to G-box motif (CACGTG). By the presence of canonical G-box motif, we identified a set of genes that influence defense response and are likely to be targets of MYC2. Out of the identified genes, we selected ICS1, MKK4, PEPR1 and RIN4 for further studies.The transcript level of these genes were highly induced after Pseudomonas syringae in MYC2 dependent manner. By gel shift assay, we show that MYC2 protein physically associates with the target gene promoters. Experiments are underway to confirm the role of MYC2 binding in regulating transcription of target genes.
PI406 (IPPC0566) Marker assisted backcross breeding for transfer of a major QTL, qDTY1.1 determining grain yield under lowland drought into an elite Basmati rice variety “Pusa Basmati 1” Gaurav Dhawan1*, Priyanka Dwivedi1, S. Gopala Krishnan1, M. Nagarajan2, P.K. Bhowmick1, Madan Pal3 and A.K. Singh1 1Division of Genetics, 3Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India 2Indian Agricultural Research Institute, Rice Breeding & Genetics Research Centre, Aduthurai-612101, Tamil Nadu, India *Presenting author: [email protected]
Drought is a major abiotic stress limiting the production of rice. It is gaining importance in India even in traditionally irrigated ecosystems, owing to decrease in availability of irrigation water primarily due to decline in ground water level as well as fluctuation in monsoons. Pusa Basmati 1 (PB1), the first semi-dwarf high yielding Basmati rice variety is popular among the farmers even today, due to its advantages of semi-dwarf growth habit, photoperiod insensitiveness and its exceptional grain and cooking quality characters. Since, it has been bred for irrigated ecosystem with assured irrigations; it is susceptible drought, which limits both yield as well as grain quality under drought stress. Marker assisted backcross breeding (MABB) was adopted to incorporate a major QTL for grain yield under drought, qDTY1.1 from a non-basmati rice variety “Nagina 22” into Basmati rice variety PB1 for enhancing grain yield under the drought stress at reproductive stage. Foreground selection was carried out at every generation using the SSR marker RM431, linked to qDTY1.1. Additionally, at each generation stringent selection for the recurrent parent phenome, grain and cooking quality characters was carried out to identify the best plant for further backcrossing. Advanced backcross progenies in BC3F2 harboring qDTY1.1 in the genetic background of PB1 has been identified. Further testing of the developed backcross derivatives under drought will help in identifying potential lines with drought tolerance as well as other desirable traits, which will be invaluable source of drought tolerance in Basmati rice improvement.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 320 Session I : Genomics-assisted Plant Improvement
PI407 (IPPC0584) Identification of genomic regions and genes for nitrogen use efficiency in rice P. Vijayalakshmi*, T. Vishnukiran, B. Ramana Kumari, K.N. Swamy, V. Jaldhani, M. Suchendranathbabu, P. Raghuveer Rao, C.N. Neeraja and S.R. Voleti ICAR-Indian Institute of Rice Research, Rajendranagar, Hyderabad, India *Participanting author: [email protected]
Association mapping is a notable strategy for identifying genes underlying quantitative traits in plants. The main aim of the present study is to find out genomic regions/genes for NUE at different N levels viz., application of N fertilizer (N100) and without application of N fertilizer (N0) during wet (2011) and dry (2012) seasons. Total 96 rice genotypes were characterized for physiological, morphological, yield and NUE related traits. Population structure or hierarchical cluster analysis was carried with 52 SSRs markers, which resulted in two major clusters. Marker traits associations were identified with general linear model (Q+K) using Tassel 2.1 software. A total 49 markers were shown significant association with multiple traits at two N levels during two seasons. Out of which, 37 markers are common for both the seasons resulting eight and four specific markers for wet and dry season. In wet season, 30 markers were significantly associated in both N levels with 9 and 6specific markers in N100 and N0, where as in dry season, 21 markers were significantly associated in both N levels with six and fourteen specific markers in N100 and N0 respectively. Finally, total ten markers were shown significant association in both N levels and seasons. Many marker associated genomic regions were coincided with already known genes and also predicted some of candidate genes for NUE with in the window of 1Mb marker region. The findings from the present study will help to identified and also provide a basis for breeding genotypes that are tolerant to low-N conditions.
PI408 (IPPC0604) RNA-Seq analysis for the study of global transcriptome in rice under induced osmotic stress Narottam Dey* Centre for Biotechnology, Visva-Bharati, Santiniketan-731235, West Bengal, India *Presenting author: [email protected]
High-throughput mRNA sequencing (RNA-Seq) offers a number of significant information of which discovery of new genes and measurements of transcript expression in a single assay is the most important one. The most crucial steps involving RNA-Seq analysis are the appropriate treatment of biological samples, sufficient biological replica, quality control of reads, proper alignments, and setting up of downstream statistical analysis. NCBI-SRA database is a valuable global resource for raw transcripts, generated through recent sequencers. The SRA data have been used to validate experimental results, determine variance and open up new avenues of Bioinformatics research. Rice, a model crop with known genome sequence has been used in a number of RNA-Seq experiments has an enriched SRA data base linked with induced osmotic stress experiments. In the present work, a comparative Bioinformatics analysis was carried out among a set of trascriptome data available in NCBI-SRA database, conducted under varied induced osmotic stresses on different rice tissues. In analysis pipeline, the raw sequence data collected in from SRA were transformed into reads (using fastqdump), quality checked (using fastqc), aligned with rice reference genome (using bowtie2 and Tophat2), assembly of transcripts (with Cufflinks and Cuffmerge), differential expression study (with Cuffdiff) and final visualization through CummeRbund. In a parallel pipeline, SRA database generated sraRunInfo.csv files were used for samples. RData which was used in a downstream analysis in R using DESeq and edgeR packages. Finally, the information generated from two parallel pipelines was compared to generate a final expression profiles with associated information.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 321 Session I : Genomics-assisted Plant Improvement
PI409 (IPPC0679) A first generation HapMap and diversity analysis in a collection of 129 cultivars of chickpea Annapurna Chitikineni1*, Manish Roorkiwal1, Weiming He2, Pooran Gaur1, Dadakhalandar Doddamani1, Jianbo Jian2, Abhishek Rathore1, Rachit K. Saxena1, Narendra Singh3, S.K. Chaturvedi4, Swapan K. Datta5, Gengyun Zhang2, Jun Wang2 and Rajeev K. Varsheny1 1International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India 2Beijing Genomics Institute (BGI) - Shenzhen, China 3All India Coordinated Research Project on Chickpea (AICRP), Indian Council of Agricultural Research, Kanpur, India 4Indian Institute of Pulses Research (IIPR), Indian Council of Agricultural Research (ICAR), Kanpur, India 5Visva-Bharati, Santiniketan, India *Presenting author: [email protected]
Chickpea (Cicer arietinum L., 2n=16) is the second most important food legume crop in the world. It contributes to a larger part of human food and animal feed in developing countries. Due to exposure of the crop to a range of biotic and abiotic stresses, the crop productivity, however, is <1 ton per hectare. Furthermore, because of a very narrow genetic diversity in the elite genepool, molecular breeding approaches have also not been in routine for crop improvement in chickpea. With an objective to estimate variations and linkage disequilibrium (LD) in elite germplasm, a non-redundant set of 129 varieties released in 12 countries (Australia, Bangladesh, Spain, Canada, USA, Ethiopia, India, Nepal, Kenya, Tanzania, Myanmar and Sudan) were subjected to whole genome re-sequencing (WGRS). While 5X-12X (with an average 7.3X) coverage WGRS data were generated for 129 lines. Alignment of sequence data for these cultivars with the draft genome sequence of chickpea has provided >1.3 million SNPs in genome, >108 K in mRNA and >42 K in CDS regions. By using these structural variations (SNPs, Indels, PAVs, CNVs), a comprehensive analysis has been undertaken on LD decay and temporal diversity in terms of released varieties in different decades, as well as market type (desi and kabuli). In addition, a first generation HapMap for chickpea has been developed that provides a resource of large-scale variations for facilitating chickpea breeding.
PI410 (IPPC0701) Characterization and development of marker for discriminating waxy and non-waxy rice germplasm Gayle Kharshiing* and N.K. Chrungoo Centre of advanced studies, North Eastern hill University, Shillong-793022, Meghalaya, India Presenting author: [email protected]
Rice starch consists of amylopectin and amylose, the latter component being controlled by the Waxy (Wx) gene. Apparent Amylose Content (AAC), regulated by the Waxy gene, represents the key determinant of rice cooking properties. Studies pertaining to the waxy locus are not only of interest in understanding gene regulation but are also important for agricultural applications. Waxy varieties of the diploid cereal rice (Oryzae sativa L.) have endosperm starch granules lacking detectable amylose. In this study, the effectiveness of available molecular markers and comparative studies was conducted in predicting the AAC values in which 70 rice accessions were characterized into waxy and non-waxy using iodine staining of starch granules. Two alleles Wxa and Wxb were found to regulate the quantitative level of the Wx protein as well as the amylose content. A comparative sequence analysis of the locus revealed the unique 23 bp duplication event which is associated with the base substitution found in the 5’intron1 splice site found in the waxy locus which discriminates the waxy from the non-waxy. The relationship of the 5’intron1 splice site with the 23 bp duplication event can further validate their association with the waxy phenotype. Further, a marker was developed to distinguish between waxy and non-waxy in rice germplasm based on the 23 bp duplication.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 322 Session I : Genomics-assisted Plant Improvement
PI411 (IPPC0729) Prediction of breeding values using genome wide markers for yield related traits in chickpea Manish Roorkiwal1*, Abhishek Rathore1, Roma Das1, Muneendra Singh1, Samineni Srinivasan1, Pooran M. Gaur1, Chellapilla Bharadwaj2, Shailesh Tripathi2, Yongle Li3, John M. Hickey4, Aaron Lorenz5, Tim Sutton6, Jean-Luc Jannink7 and Rajeev K. Varshney1 1International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India; 2Indian Agricultural Research Institute (IARI), Delhi, India; 3Australian Centre for Plant Functional Genomics (ACPFG), University of Adelaide, SA, Australia; 4The Roslin Institute, The University of Edinburgh, Scotland, UK; 5University of Nebraska, Lincoln, USA; 6South Australian Research and Development Institute (SARDI), SA, Australia; 7Cornell University, Ithaca, USA *Presenting author: [email protected] Genomic selection (GS) unlike MABC (marker assisted backcross) predicts breeding values of lines and makes selection prior to phenotyping using genome-wide marker profiling. To address the low productivity in chickpea owning to several biotic and abiotic stresses, a collection of 320 elite breeding lines was selected as the “training population”. Training population was phenotyped extensively for yield and yield related traits at ICRISAT, Patancheru and IARI, Delhi during crop season 2011-12 and 2012-13 under rain-fed and irrigated conditions. In parallel, the training population was genotyped using DArTseq arrays (15,360 features). Collected phenotypic data and generated genome-wide marker profiling data were used with six statistical models including RR-BLUP, kinship based ridge regression, BayesCπ, BayesB, Bayesian LASSO and random forest to predict genomic estimated breeding values (GEBVs). GS models were tested for four yield related traits namely seed yield, 100 seed weight, days to 50% flowering and days to maturity. Prediction accuracy for the models tested varied from 0.912 (SDW) to 0.138 (seed yield). Heat map analysis using genotyping data to understand the relationship within these lines suggested possibility of two different groups similar to cluster analysis. In order to understand the effect of population structure on accuracy, analysis was re-performed by implementing population structure for calculation of GEBV. Population structure did not have significantly effect on prediction accuracy. Results obtained are encouraging for deploying GS in chickpea breeding.
PI412 (IPPC0753) In silico analysis of an endosperm-specific legumin-type seed storage protein gene and 5’UTR of common buckwheat (Fagopyrum esculentum Moench) Lashaihun Dohtdong1*, Kamlesh Kant Nutan2, Nikhil Kumar Chrungoo1 and Ashwani Pareek2 1Department of Botany, North Eastern Hill University, Shillong-793022, India 2Stress Physiology and Molecular Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected] The nutritional quality of food and its availability are the key factors to food security. We report the cloning of a full length 1.7 kb gene (acc. no. KM488332) coding for a legumin type seed storage protein of common buckwheat (Fagopyrum esculentum Moench). BLAST analysis of the sequence revealed >90% homology with legumin gene nucleotide sequences bearing accession numbers D87980, GQ358524 and AY245536.The deduced amino acid sequence from the open reading frame code for a 64 kDa preprotein. BLASTP analysis of the sequence revealed 98% homology with 13S globulin seed storage protein from common buckwheat (acc.no.O23878). Analysis of the deduced amino acid sequence revealed the presence of an α and a β subunit linked together ASN-GLU linkage with the β subunit showing 5.57% lysine, which is amongst the highest recorded for legumin type proteins. Using the gene walking approach, we have isolated the 1kb 5’UTR (acc. no.EU595873) of the gene as identified by GENSCAN 1.0. NNPP predicted three probable transcription start sites out of which the TSS at P’733 is located closest to the predicted ATG start codon (P’774) and follows the YR rule. Other cis-elements identified in the sequence included the -30 TATA box, an endosperm specific motif identified as prolamin box in cereals ‘TGTAAAG’, three RY elements ‘CATGCA’, core of ‘AACA’ motif for regulating endosperm specific and quantitative expression, CAAT’ promoter consensus sequence and ‘CACA’ box.We suggest a strong potential for the gene and its promoter in enhancing the nutritional levels of lysine in conventional crops such as rice through transgenic approaches.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 323 Session I : Genomics-assisted Plant Improvement
PI413 (IPPC0801)
Inheritance of red kernel colour in rice (Oryza sativa L.) and genotypic screening of F2 population through molecular markers Ashish Jondhale, Sanjay Bhave*, Jalindar Devmore, Murad Burondkar and Vijay Dalvi Dr. B.S. Konkan Krishi Vidyapeeth Dapoli-415712, Maharashtra, India *Presenting author: [email protected]
Inheritance of red kernel colour in rice (Oryza sativa L.) and genotypic screening of F2 population through molecular markers was studied in two crosses involving one white kernel and another red kernel parents. Result indicated that the red kernel colour of rice was governed by a single dominant gene. Interactions confirmed by making testcrosses of the F1s with the white kernel parents which were recessive for the kernel colour. In molecular screening, 13 SSR markers linked to the red kernel colour trait were utilized and among these, two SSR markers (RM 206 and RM 251) showed parental polymorphism. A representative 100 individuals derived from the crosses Karjat-2 x Munga and their reciprocal was estimated for white and red kernel colour. Estimation of nutritional aspects especially micronutrients viz; iron, zinc, manganese and copper exhibited wide variation in both, white and red kernel parents and their hybrids. The iron, zinc, manganese and copper contents in the red rice were found to be higher than that of white rice. It clearly indicated that red rice was highly nutritive than that of white rice. Beside this, generation mean analysis, revealed the significant Mather’s individual scaling tests and joint scaling test for all the characters in both the crosses. Among non allelic gene interactions, additive x additive and dominant x dominant were more prominent and played significant role in inheritance of the several traits studied. The additive x dominant interaction were significant but had lesser preponderance for governing the traits.
PI414 (IPPC0829) Isolation and characterization of UDP-glucose pyrophosphorylase (UGPase) promoter from potato (Solanum tuberosum L.) Shashank Yadav1*, Lekshmy S.1, Rakesh Verma1, Santosh Vinjamuri1, Monika Dalal2 and Viswanathan Chinnusamy1 1Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India 2National Research Centre on Plant Biotechnology, New Delhi-110012, India *Presenting author: [email protected]
Low temperature storage of potato is inevitable to prevent sprouting and diseases, retention of dry matter and extended marketability. However, Cold Induced Sweetening (CIS) is a problem associated with cold storage. Towards unravelling the genotypic differences in CIS, we analyzed allelic variation in genes involved in CIS, and characterized UGPase promoter. UGPase catalyzes reversible production of UDPG and pyrophosphate (PPi) from Glc-1-P and UTP. UGPase expression was analyzed in potato genotypes with contrasting CIS. Previously reported allelic variations in UGPase, apoplastic invertase and invertase inhibitor were analyzed in Indian potato genotypes. Absence of UGPase “B”, apoplastic invertase inhibitor StINHap-a and apoplastic invertase Stm1051-255 alleles, and presence of apoplastic invertase Stm1051- 221 were associated with CIS tolerance of potato. Expression of UGPase gene was highly induced by cold storage in CIS tolerant genotypes Kufri Chipsona 1 and Kufri Frysona, whereas the expression was low in CIS sensitive genotype Kufri Pukhraj. To unravel the differential expression of UGPase genes between CIS tolerant Kufri Chipsona 3 and CIS sensitive Kufri Pukhraj, the promoter of UGPase was cloned and sequenced. These two genotypes differ by 49 and 6 base InDels in UGPase promoter. Based on this InDel, a PCR marker was developed and validated in 38 potato genotypes. CIS tolerant genotypes showed 55 bp deletion in UGPase promoter. Identification of trans-acting factors and cis-elements involved in cold-induction of CIS genes will further illuminate the mechanisms of CIS tolerance.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 324 Session I : Genomics-assisted Plant Improvement
PI415 (IPPC0865) Next generation transcriptome sequencing to elucidate molecular mechanism of Self-incompatibility in tea Romit Seth*, Aparna Shree Raina, Abhishek Bhandawat, Rajni Parmar, Gagandeep Singh and Ram Kumar Sharma CSIR-Institute of Himalayan Bioresource Technology, Palampur-176061, (H.P.), India *Presenting author: [email protected]
Self-incompatibility is a genetically controlled mechanism existing in higher plants, especially in angiosperms and plays critical role to inhibit inbreeding depression, henceforth, enhancing heterozygosity, resulting in diversification and speciation of plant species. Tea (Camellia sinensis (L) O. Kuntze), being an obligate outcross is highly heterogeneous thus hindering the maintenance and propagation of quality lines for commercial cultivation. To unravel the complexity of this trait, whole transcriptome sequencing of self-incompatible and cross-compatible samples were performed using Illumina platform. In total, 98.5 million raw reads were generated followed by de novo assembly of 73.3 million clean reads resulted 26Mb non- redundant data (48,893 contigs). Functional annotation identified 32,563 unique contigs using sequence search against KEGG, GO, SWISSPROT, TAIR, KOG, Transcription Factor Database. This study revealed that contigs encoding S- Locus and apoptosis related genes were found to be differentially expressed, which might be playing role in regulating the pathways of self-incompatibility. This trait specific study will help in predicting the possible molecular pathways responsible in providing the self-incompatibility in tea. It may be used for future genomic studies, which in the downstream, will help in maintenance and propagation of quality lines in tea.
PI416 (IPPC0867) Global transcriptome analysis for dissecting drought tolerance in tea Rajni Parmar*, Abhishek Bhandawat, Gagandeep Singh, Kuldip Jayaswall and Ram Sharma Biotechnology Division, CSIR, Institute of Himalayan Bioresource Technology, Palampur-176061, Himachal Pradesh, India *Presenting author: [email protected], [email protected]
Abiotic stresses are environmental conditions that reduce growth and yield of crop below optimum level and especially, drought stress is primary cause of crop loss worldwide. Tea [Camellia sinensis (L.) O. Kuntze] is perennial evergreen plant of genus Camellia belongs to Theaceae family. Drought stress is one of severe abiotic stress that limits growth, yield and geographical distribution of tea. Field evaluation studies over the years identified potential source of drought tolerance in tea. Interestingly, these drought tolerant genotypes capture moderate to high level of diversity. Although, few molecular studies on drought have been conducted in the recent past but genome wide transcriptomic studies have not been undertaken so far. Moreover, multiple genotypes having recorded different response to drought (tolerant/susceptible) have not been studied in tea. During this study, sequencing of five diverse genotypes having background knowledge of drought response have been done with induced drought at the transcriptome level. A total of 116,666,747 filtered read were generated. After de novo assembly 36,216 contigs were obtained with an average length of 847 bp. Analysis of these contigs using non-redundant protein database annotated 28,951 unigenes showed number of genes have significant role in drought stress. Additionally 2,42,846 high quality SNPs were ascertained from this dataset. Present study will help to elucidate the drought responsive key molecular players and also in gaining deeper understanding of behavior of diverse genotypes of tea under drought stress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 325 Session I : Genomics-assisted Plant Improvement
PI417 (IPPC0898) Transcriptome sequencing, genome-wide miRNA prediction and their target identification from horsegram accessions with contrasting drought tolerance Jeshima Khan Yasin1*, Vivek Thiruvettai, M. Arumugam Pillai2, K.V. Bhat1, Viswanathan Chinnusamy3 and K.C. Bansal1 1National Bureau of Plant Genetic Resources, New Delhi-110012, India 2Department of Plant Breeding, Killikulam, Tamil Nadu Agricultural University, Tamil Nadu, India 3Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Horsegram (Macrotyloma uniflorum) is an unexploited legume with high adaptability to abiotic stresses. A total of 3300 accessions of horsegram from national gene bank were phenotyped and 50 accessions with contrasting drought tolerance were identified. Two accessions with contrasting reactive oxygen scavenging potential were selected and subjected to drought stress. Transcriptome data were generated for control and stressed plants. De-novo transcriptome annotation identified genes for functional categories such as stress–related (2.87%), signalling (4.31%), hormone metabolism (2.1%) protein metabolism (13.64%) and RNA metabolism (8.33%). Confirming our earlier finding that horsegram employs energy conservation mechanisms, which were explained by transcript abundance for glycolysis (0.47%), gluconeogenesis/ glyoxylate cycle (0.06%), TCA (0.38%), and mitochondrial electron transport/ATP synthesis (0.55%). Annotation of horsegram transcriptome was helpful in determining the interplay of multiple metabolic activities involved in stress responses as compared with soybean. Non-coding and weak-coding sequences were analysed for miRNAs based on their secondary structure and drought stress responsive miRNAs were identified. The targets for the miRNAs were predicted. miRNAs from other seven related legumes were collected from miRBase to find their targets in horsegram transcriptome. Predicted stress responsive miRNAs were confirmed to regulate drought tolerance and validated in other legume crops. The novel miRNAs predicted in this study needs further validation. Reference based genome mapping was performed using Glycine max as reference genome to identify likely QTLs, candidate gene based markers, exon junctions, etc. The genomic resources generated in this study will be highly useful in genetic improvement of horsegram and related pulses.
PI418 (IPPC0917) Prospecting candidate genes associated with abiotic stress tolerance from proteins of unknown function (PUFs) identified from Indian mulberry (Morus sp.) K.H. Dhanyalakshmi*, R.S. Sajeevan and Karaba N. Nataraja Department of Crop Physiology, University of Agricultural Science, GKVK, Bangalore-560065, Karnataka, India *Presenting author: [email protected] Abiotic and biotic stresses have become serious challenges to crop production. Plant growth, development and responses under these conditions are tightly regulated by the coordinated expression of diverse molecular, biochemical, cellular and physiological events that help them to grow and survive. At molecular level, all stress adaptive processes are mainly mediated by the expression of several stress responsive genes, their products and pathways and the roles of some of such genes have been well characterized. But, a vast majority of proteins in public databases still remain uncharacterized or unknown or hypothetical, collectively called as proteins of unknown functions (PUFs) which might be novel and unique. Understanding their relevance might help to address complex mechanisms of stress adaptation in a better way. In this context, an attempt was made to validate the role of selected PUFs identified from drought transcriptome (SRA accession SRP047446) of Indian mulberry. The PUFs were uncharacterized according to Uniprot blast analysis. We used diverse sequence as well as structure level computational analysis tools to assign probable function to over 100 selected PUFs. The response of a few selected PUFs to different abiotic stresses was examined under laboratory conditions in mulberry. The selected PUFs were up-regulated under these stresses and further validation might uncover novel pathways associated with stress tolerance in mulberry.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 326 Session I : Genomics-assisted Plant Improvement
PI419 (IPPC0920) Expression analysis of genes involved in stem reserve accumulation and mobilization in rice under soil moisture deficit Puja Rai1*, A.S. Sakhare1, Chandarpal Vishwakarma1, P.C. Nautyal2 and Viswanathan Chinnusamy1 1Division of Plant Physiology, 2Division of Seed Science and Technology, Indian Agricultural Res. Institute, New Delhi-110012, India *Presenting author: [email protected]
Starch is the major storage carbohydrate in rice and is stored in the stem tissues when photosynthates production is higher than sink demand. The stored non-structural carbohydrates (starch and sucrose) provide an alternative source during later stage of grain filling or under abiotic stresses when current photosynthesis is reduced. Genes coding for starch biosynthesis and degradation, sucrose synthesis and sucrose transport have been studied in mainly in cereal grains. However, their regulation and involvement in stem reserve accumulation and mobilization under soil moisture deficit stress is poorly understood. This study was aimed to understand the regulation of genes involved in stem reserve accumulation and mobilization in rice and their regulation under soil moisture deficit stress. Real time RT-PCR expression analysis was performed to study the gene expression in different tissues namely leaf, leaf sheath and stem of rice cv. Pusa Sugandh 2 under control and drought stress conditions. The results identified specific members of AGPase, starch synthase and sucrose transporters (SUT) gene families that predominately express in stem and their regulation under drought stress. Expression of genes involved in stem reserve accumulation and mobilization under control and drought conditions in two contrasting rice cultivars is being studied. Understanding of the regulation of genes involved in stem reserve mobilization under drought will open up new possibilities to select high yielding drought tolerant rice varieties in future.
PI420 (IPPC0944) Introgression of diverse physiological traits to improve drought adaption in rice by marker assisted multi-parent backcross breeding strategy M.D. Prathibha1*, M.V. Mohan Kumar1, N.M. Mallikarjun2, B.R. Raju1, D. Pushpa1, H.G. Jalendra Kumar1, K. Sumanth Kumar1, J. Smitharani1, M.P. Rajanna3, M. Udayakumar1 and M.S. Sheshshayee1 1Department of Crop Physiology, 2Department of Genetics and Plant Breeding, University of Agricultural Science, GKVK, Bangalore- 560065, Karnataka, India; 3ZARS, VC Farm, Mandya-571401 *Presenting author: [email protected]
With the rapidly decreasing water resources, devising water saving agronomic practices for rice cultivation is the most crucial need of the hour. Global opinion strongly favours a trait based strategy over selection for absolute yield under stress in crop improvement. Traits associated with maintenance of water relationships and superior metabolism under water limited conditions deserve emphasis. Therefore water mining traits (root), efficient use of water and Cellular Level Tolerance (CLT) have greatest relevance. In an earlier investigation, a panel of germplasm was characterised for molecular and phenotypic diversity leading to the discovery of DNA markers by Association Mapping and trait donor genotypes (AC- 39020 for roots and CLT, IET-16348 for WUE). We initiated a focused marker assisted backcross breeding to pyramid these complex physiological traits into the elite genetic background of IR-64. Of the markers discovered earlier by association mapping, 20 polymorphic markers were used for foreground selection and a genome wide set of 120 polymorphic markers were used to reconstitute the recurrent parent genome. Of the 10000 DCBC3F2 lines generated, a set of 1500 lines were screened under aerobic field conditions and the best lines (250) were advanced to DCBC3F3. These lines were extensively phenotyped for root traits and Δ13C to verify trait introgression. This study significantly contributed for the development of trait introgressed lines that can be used for commercial cultivation and QTL-NILS that have great potential in academic understanding of QTL interactions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 327 Session I : Genomics-assisted Plant Improvement
PI421 (IPPC0945) Study of molecular basis of heterosis revealed dominant and additive gene expression patterns in Capsicum hybrids Sushil Chhapekar*, Rashmi Gaur, Mukesh Nitin, M. Sarpras, Vineet Sharma and Nirala Ramchiary School of Life Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Capsicum species commonly known as chilli peppers are economically important vegetable and spice crop of the world. Chilli fruits are rich source of vitamins, antioxidants and nutrients along with anti-cancer and several medicinal properties. India is the world’s largest producer, consumer and exporter of chilli peppers; however, the yield potential is very low due to poor yielding varieties and high incidence of pests and diseases. One of the methods to achieve quantum jump in yield and quality is through heterosis breeding. Although heterosis has been widely exploited in other crops, the study of molecular and genetic mechanisms underlying the superiority of F1s over parental lines are poorly understood in Capsicum plants. An attempt has been made to dissect the molecular basis of intra (Capsicum chinense X C. chinense) and inter- specific heterotic F1 hybrids (C. chinense X C. frutescens and C. annuum X C. chinense) in capsicum species. The interspecifc F1 hybrids were observed more heterotic compared to intra-specific hybrids in overall plant phenotype except for fruit traits including plant growth (height and area), vigor, chlorophyll content, and other metabolites. Transcriptome sequencing of early heterotic F1 plants showed more dominant and additive gene expression differences compared to parental lines which might be the possible reason(s) for superiority of performance of heterotic F1 hybrids. Detailed functional analysis of genes showed that most of the differentially expressed genes belong to photosynthetic pathways, carbohydrate metabolism, hormonal signalling and code for transcription factors. Some of the differentially expressed genes are being validated which might be helpful for designing effective breeding strategies to enhance capsicum productivity.
PI422 (IPPC0951) Discovery of QTLs and validation of markers for drought adaptive traits through characterization of trait specific mapping population in rice (Oryza sativa L.) Pooja Bharti1*, B.R. Raju1, R. Ramachandra, M.P. Rajanna2 and M.S. Sheshshayee 1Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore-560065, Karnataka, India 2AICRIP (Rice), ZARS, V.C. Farm, Mandya-571405, Karnataka, India *Presenting author: [email protected]
Introgression of physiological traits like water mining and WUE onto a single elite background is one of the most appropriate strategies to improve productivity under water limited conditions. Accelerating such trait introgression can only be achieved by the adoption of focused molecular breeding practices. Identification of DNA markers associated and/or linked with such complex physiological traits is therefore the primary pre-requisite. Towards identifying markers and QTL, we adopted association mapping as well as bi-parental mapping strategies. A set of 230 F8 RIL population developed by crossing IET 15963 (high root) and Thanu (high WUE) was extensively phenotyped for root, WUE traits and spikelet fertility under aerobic conditions. A linkage map consisting of 120 linked markers was constructed and robust QTL regions governing root, WUE, spikelet fertility and other drought adaptive traits. The bi-parental mapping led to the validation of markers for WUE and root traits previously discovered by association mapping. Several trait introgressed transgressive segregants were identified and were characterized for field level performance under aerobic conditions.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 328 Session I : Genomics-assisted Plant Improvement
PI423 (IPPC0953) Strategy for Meta QTL analysis by phenotyping diverse trait specific mapping populations for drought adaptive traits in rice (Oryza sativa L.) J.A. Smitharani1*, M.S. Sheshshayee1, B.R. Raju1, M. Raveendran2, S. Robin2, N. Sarala3, M.D. Prathiba1 and M. Udayakumar1 1Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore-560065, India; 2Tamil Nadu Agricultural University, Coimbatore-641003, India; 3Indian Institute of Rice Research, Hyderabad, India *Presenting author: [email protected] Emperical selection for drought adaptive traits is extremely complex which slows down crop improvement. This led to a spurt of research attempting to discover QTL. Though marker assisted breeding has the potential in accelerating crop improvement, the diverse traits used by investigators and the diverse genetic background of populations used has significantly complicated the process of breeding. In recent years increased efforts are being made to validate QTL by identifying consensus regions by “Meta QTL” analysis. In these analyses published data on similar traits across various genetic backgrounds are considered to identify common genomic regions, governing variability in a specific trait. In this study, we report a strategy to identify consensus genomic regions conferring variability in physiological traits associated with drought adaptation in rice. The parental genotypes that have been used in developing mapping populations were initially phenotyped to assess variability in drought adaptive traits like roots and WUE. Specific mapping populations from contrasting parents were identified. RILs developed from IR 64 x Norungan, IR 64 x Kallurandaikar, Swarna x Madhukar and Thanu x IET 15963 were extensively phenotyped for drought adaptive traits. Root structure experiments were conducted to assess variability in root traits and the Delta 13 C was used to measure WUE. More than 1500 QTL reported from our Centre and from elsewhere were used to initially determine the parental polymorphism. Selective genotyping of the contrasting groups of RILs differing in specific traits would lead to a more precise Meta QTL analysis and validation of markers for marker assisted breeding. PI424 (IPPC0985) Physiological characterization of an upland rice Nagina-22 derived EMS mutants–contrasting for leaf size H.R. Sowmya1,2*, K. Sumanth Kumar1, Rajesh Kumar Singal1, P. Niranjana2, M.P. Rajanna3, R.P. Sharma4, T. Mohapatra5, N. Sarala6, S. Robin7, A.K. Singh8 and M.S. Sheshshayee1 1Department of Crop Physiology, University of Agricultural Sciences, GKVK, UAS Bengaluru-560065, Karnataka, India; 2Department of Biochemistry, Kuvempu University, Shivgoga; 3Zonal Agricultural Research Station, College of Agriculture, Mandya; 4National Research Centre on Plant Biotechnology, IARI, New Delhi-110012, India; 5Central Rice Research Institute, Cuttack-753006, India; 6Directorate of Rice Research, Rajendranagar-500030, Hyderabad, India; 7Department of Rice, Tamil Nadu Agricultural Research Institute, Coimbatore, India; 8National Bureau of Plant Genetic Resources, New Delhi-110012, India *Presenting author: [email protected] A greater understanding of the molecular basis of regulation of canopy architecture has great relevance in determining growth rates. In this investigation, we report the identification of EMS induced mutants of rice in the background of Nagina-22 with significant differences in leaf size and their physiological characterization. The narrow leaf mutant
(N22_BADT_380_4) showed lower stomatal conductance (gs) than broad leaf mutant (N22_BADT_437_1) while photosynthetic rate was comparable. This influence was verified by comparing whole plant cumulative water transpired (CWT). The narrow leaf mutant recorded lower CWT than broad leaf mutant and Nagina 22 (wild type). On the other hand, the narrow leaf mutant would maintain relatively higher gs and A under stress, while a significant reduction was noticed in broad leaf mutant and the wild type when the plants experienced moisture stress. The leaf thickness as indicated by SLW was significantly higher in narrow leaf mutant which also recorded higher epicuticular wax load (EWL). With an increased mesophyll cell number combined with reduced heat load because of EWL, the narrow leaf mutant was capable of maintaining higher photosynthetic rate per unit area. However, because of a significant reduction in tiller number, the total canopy leaf area was less and hence TDM of narrow leaf mutant was also less. These leaf size mutants represent an excellent resource for generating greater clarity on the factors contributing to canopy architecture and canopy carbon exchange rate in rice.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 329 Session I : Genomics-assisted Plant Improvement
PI425 (IPPC0996) RNASeq study to understand molecular basis of rice- Rhizoctonia interaction Archana Kumari1*, Priyanka Jain2, T.R. Sharma2 1Division of Plant Physiology, IARI, New Delhi-110012, India 2National Research Centre on Plant Biotechnology, Pusa Campus, New Delhi- 110012, India *Presenting author: [email protected]
Sheath blight, caused by Rhizoctonia solani, is one of the most serious diseases of rice next to rice blast. For understanding plant pathogen interaction, advanced genetic and statistical methods of analysis have been used in the present study to identify new resistance genes through transcriptional profiling. RNA Seq analysis of resistant and susceptible rice lines after infection with R. solani was performed at 0, 12, & 24 hours post infection. RNA Seq data revealed many differentially expressed genes at 12 hrs and 24 hrs post infection. We obtained 84 (78 upregulated & 6 downregulated), 298 (200 upregulated & 98 downregulated) and 196 (175 upregulated & 21 downregulated) significant (FDR adjusted p value <0.05) differentially expressed locus (SDEL) in resistant rice line at 0, 12 & 24 hpi respectively. In comparison we could get 342 (335 upregulated & 7 downregulated), 231 (146 upregulated & 185 downregulted) and 873 (632 upregulated & 241 downregulated) SDEL in susceptible rice lines at 0, 12 & 24 hpi, respectively. Gene ontology (GO) annotation and pathway analysis of SDEL showed that genes involved in signalling, transcription factors, response to stress and defense related genes are enhanced in resistant line compared to susceptible one at 12 & 24 hpi. We report most significant changes in gene expression in rice sheath for the first time and reveal the genes associated with various pathways and processes in plants during infection. Present study of rice- Rhizoctonia interactions have been undertaken to uncover the mechanisms by which disease resistance is achieved.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 330 Session J : Flowering and Post Harvest Physiology
PJ426 (IPPC0239) Deciphering the biochemical basis of flower colour variation in palas (Butea monosperma Lam) taubert Vaibhav D. Lohot1*, A. Roy Chowdhury2, Nandkishore Thombare2, Jyotirmoy Ghosh1, Thamilarasi K1, A Mohanasundaram1, Gunjan1 and Kewal K. Sharma1 1Lac Production Division, IINRG, Ranchi-834010, Jharkhand, India 2Process and Product Development Division, IINRG, Ranchi-834010, Jharkhand, India *Presenting author: [email protected]
Angiosperms exhibits a tremendous diversity of flower colours, with sister species often differing in the intensity, hue or patterning of the corolla. This flower colour trait is of paramount importance in the ecology and evolution of plants due to its ability to attract pollinators. The colouration of flowers is due to accumulation of flavonoids (including anthocyanins). Palas (B. monosperma) belongs to the family Papilionaceae, enduring beautiful scarlet (orange-red) coloured flowers during early summer. In Jharkhand Palas is extensively utilized for lac cultivation and is the single largest producer of rangeeni strain of Indian lac insect (Kerria lacca Kerr). Occasionally in Palas different shades of flower colours ranging from brick-red to pure white with several shades of intermediate orange-yellow and yellow colours are seen in wild. An attempt was made to decipher the basis of colour variation in Palas flowers by quantitative estimation of flavonoids and anthocyanins. Variants were classified into three categories based on colour similarity viz. orange-yellow, yellow and scarlet. The study revealed that negative values were obtained for anthocyanin production in yellow coloured flowers regardless of flavonoid production which indicates that plant is unable to convert flavonoids to anthocyanins. In orange- yellow variants, the conversion of flavonoids to anthocyanins was ranged from 35-60 percent. Though the scarlet produces maximum flavonoid content among all the variants, conversion percentage recorded very low. Thus the study revealed that intensity of the flower colour variation in Palas could be due to interplay of flavonoid content and its extent of conversion to anthocyanins.
PJ427 (IPPC0254) Functional characterization of flowering inducer FT-like gene family in banana (Musa acuminata L.) Akhilesh Kumar Chaurasia1*, Hemant Patil1, Bal Krishna1, V.R. Subramaniam1, A. Sane2 and P.V. Sane1 1Plant Molecular Biology Lab, Agri Park, Jain Hills, Shirsoli Road, Jalgaon–425001, India 2Plant Gene Expression Lab, CSIR-National Botanical Research Institute, Rana Pratap Marg, Lucknow-226001, India *Presenting author: [email protected]
The phase transition from vegetative to reproductive stage is an important developmental step in the life of flowering plants. It is controlled by exogenous factors such as photoperiod, temperature and abiotic stresses and endogenous factors such as hormone levels and C/N ratio. The key regulator of flowering is FLOWERING LOCUS T (FT), a member of phosphatidylethanolamine-binding protein (PEBP) super family. The FT protein, believed to be the elusive ‘florigen’, is a mobile protein which moves from the leaf to the shoot apex through phloem where it interacts with another protein FD to activate downstream targets LEAFY and APETALA1 which are responsible for converting the apical meristem to an inflorescence meristem. Phase transition/flower induction in ever green herbaceous perennials like banana is not known. In this study, we have identified and characterized 12 genes, designated as MaFT1-12, from the FT-like group of the PEBP family in banana (Musa acuminata L. AAA group cultivar Grand Nain). Time-course dependent expression during the ten months of plant growth suggests an increase in expression of MaFT2-5 and MaFT12 just prior to initiation of flowering. Functional analysis in Arabidopsis through over expression shows early flowering by MaFT1-5, MaFT8 and MaFT12 compared to wild type. These genes are also able to complement the late flowering ft-10 mutant with MaFT2 and MaFT5 exhibiting a stronger phenotype. Our study, the first of its kind in banana, shows that MaFT2 and MaFT5 (along with other genes) are key flowering inducers in banana.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 331 Session J : Flowering and Post Harvest Physiology
PJ428 (IPPC0278) Studies on modified atmosphere storage of spikes Gladiolus spp. Harsimran Kaur*, Kushal Singh and Seema Bedi Department of Botany, Punjab Agricultural University Ludhiana-141013, Punjab, India *Presenting author: [email protected]
The present study was undertaken to find out the storage response of spikes of different cultivars of gladiolus under modified atmosphere storage. The spikes of four cultivars of gladiolus viz. Jacksonville Gold, White Prosperity, Sancerre and Nova Lux were harvested when coloured tepals were visible in 6 florets per spike. The spikes were grouped into bundles of three each and packed in polypropylene (PP) and low density polyethylene (LDPE) film sleeves (25 μ thick). The sleeves were hermetically sealed at both ends. The packages were subjected to modified atmosphere (MA) storage in a cool chamber (4±0.50C; 90-95 % R.H.) for 3, 6, 9, 12, 15 and 18 days, respectively, thereafter vase life was studied at room temperature. Vase life and percent opening of florets showed decline with the progress of storage duration in all the four cultivars. Cultivar Jacksonville Gold took minimum number of days (2.40) for opening of the basal floret while the cv. White Prosperity took maximum number of days (3.14) for opening of the basal floret. Vase life and percent opening of florets were also maximum in cv. Jacksonville Gold. Spikes stored in PP packages exhibited slightly higher vase life and floret opening than those stored in LDPE packages. The spikes exhibited considerable increase in peroxidase and catalase activities in the tepals during storage, the increase being more prominent in cv. Jacksonville Gold. The increase in activity of these enzymes indicates the activation of antioxidant enzyme system to alleviate oxidative stress during the refrigerated storage.
PJ429 (IPPC0377) Regulatory evolution in homeologs of suppressor of overexpression of constans1 and its role in floral transition Tanu Sri*, Pratiksha Mayee, Shikha Tyagi, S.M. Shivaraj, Anupama Atri and Anandita Singh Department of Biotechnology, TERI University, New Delhi-110070, India *Presenting author: [email protected]
Successful floral transition of an angiosperm is achieved by a complex interplay of signalling cascades involving multiple flowering time genes. This study is focussed on a floral pathway integrator SOC1, a MADS-box transcription factor. We have examined its evolution and characterised its role in flowering in Indian Brassicas. Microsyntenic status of Brassica rapa regions containing three homeologs of SOC1 conformed to differentially fractionated homeologous sub-genomes. Screening 18 cultivars of 6 Brassica species led to identification of 8 genomic and 27 transcript variants of SOC1, including splice-forms and intron-less SOC1 genes. Phylogenetic analyses revealed the diploid base genome and sub-genome of origin of each Brassica SOC1 homologs, indicating sub-genomic organization in un-characterized Brassica genomes. Expression analyses of two B. juncea SOC1 homeologs (MF2 and LF) revealed near identical expression pattern. However, MF2-specific homeolog exhibited significantly higher expression implying regulatory diversification. To further investigate, promoters of Brassica SOC1 homologs were isolated and analysed for divergence in cis-regulatory motifs. Cumulative expression analysis of B. juncea SOC1 revealed increased levels of SOC1 transcript in various organs during flowering, highest in apical bud, establishing its role in floral transition. Furthermore, B. juncea SOC1 was functionally characterized by gain-of-function Arabidopsis mutants showing earliness in flowering. Knock-down lines of A. thaliana and Brassica juncea expressing artificial miRNA against SOC1 displayed delayed flowering along with concomitant presence of mature artificial miRNA and cleaved SOC1 transcripts. Overall, this study highlights the importance of SOC1 in floral transition in Brassica and provides insights into evolution of homeologs of developmental genes.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 332 Session J : Flowering and Post Harvest Physiology
PJ430 (IPPC0676) The bZIP transcription factor, OsbZIP, in rice is a functional ortholog of Flowering Locus D (FD) in Arabidopsis Amarjot Kaur*, Aashima Nijhawan, Mahesh Yadav and Jitendra Khurana Dept. of Plant Molecular Biology & Interdisciplinary Centre for Plant Genomics, Univ. of Delhi South Campus, New Delhi-110021, India *Presenting author: [email protected]
Flowering is an intricate pathway in plant life cycle and a plethora of genes are involved in bringing about transition from vegetative to reproductive stage. This pathway has been well characterised in Arabidopsis wherein Flowering Locus D (FD) interacts with Flowering Locus T (FT) and activate the transcription of floral meristem identity genes. However, this pathway is less understood in rice and a number of other factors are involved in flowering. Our lab identified a few bZIPs that may be involved in regulating flowering in rice employing whole genome transcriptome analysis. Among these, one of the OsbZIPs showed some promising results as it was able to complement fd mutant of Arabidopsis. In contrast to earlier reports that FD and FT interact directly, it has now been shown that their interaction is mediated by an intracellular receptor GF14-3-3 protein, which serves as a chaperone. Thus in our study too, yeast one-to-one interaction of OsbZIP (identified in this study) with OsFTLs and GF14-3-3 was performed and a few of them did show interaction. Also, yeast- three-hybrid assay involving 14-3-3 proteins showed similar results thereby suggesting the possible involvement of GF14- 3-3 protein in flowering in rice. Rice RNAi transgenics for OsbZIP showed a slight delay in flowering time in comparison to WT. These results thus provide evidence that OsbZIP might be acting as a putative ortholog of Arabidopsis FD and may be involved in regulating transition to flowering.
PJ431 (IPPC0858) Soil moisture deficit stress effect on flowering and fruit development in banana cv. Grand Naine Iyyakutty Ravi* and M. Mayil Vaganan Crop Production Section, ICAR-National Research Centre for Banana, Trichy-620102, Tamil Nadu, India *Presenting author: [email protected]
India is the largest producer of bananas. The banana crop cycle extends a minimum of one year and it requires a continuous source of soil moisture for optimum growth. Due to long crop cycle, the banana may face soil moisture deficits any time as a consequence of the seasonal decline in soil water availability. A field study was conducted to identify the effects of soil moisture deficit stress in banana cv. Grand Naine at NRCB farm. The stress was imposed by withholding irrigation at 3rd, 5th month after planting and at flowering to reach the soil moisture stress level of -0.6 to -0.7 MPa, thereafter stress was relieved by irrigation till harvest. At harvest, in irrigated control, the average bunch weight was 21.5 kg. The stress imposed at 3rd month after planting did not affect the bunch weight. However, stress imposed at 5th month after planting had a severe impact on bunch morphology, size and weight. Around 35-40% of bunches expressed choking and were malformed, and mostly weighed around 9.1 kg, with small fingers. Stress imposed at flowering resulted in poor development of fingers and became unmarketable bunches. The results revealed that irrigation at 5-6th month and at flowering was critical for normal bunch and fruit development in banana cv. Grand Naine. Foliar priming of the banana plant with acetylsalicylic acid (0.1 mM) and butylated hydroxy toluene (100 ppm) alleviated the negative effect of soil moisture deficit stress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 333 Session J : Flowering and Post Harvest Physiology
PJ432 (IPPC0982) Identification of less photo-sensitive activation tagged mutants of rice and their flanking sequence analysis to identify the possible genes Anita Pitagi1*, N.B. Shashibhushan1, Appanna V. Koppad1, Rohini Sreevathsa2, Mahantesha Naika1, B.M. Raju1 and M. Udayakumar 1Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bengaluru-560065, Karnataka, India 2National Research Centre on Plant Biotechnology, IARI, New Delhi-110012, India *Presenting author: [email protected]
Many rice genotypes with improved adaptation to abiotic stresses and more specifically, for aerobic cultivation are photosensitive. JBT36/14, an upland rice genotype adapted to aerobic cultivation has superior water relations (high yield, high roots and better WUE). However, this genotype is highly photosensitive. With an objective of identifying mutants with less photosensitivity and genes associated with it, a gain-in-function activation tagged mutants were developed in the background of JBT36/14 using activation tagging vector, pMN20 having 4x35s enhancers and glyphosate selection marker, mEPSPS. The putative mutants were identified based on the response to glyphosate and the positive mutants were screened for several phenotypic characteristics including flowering duration, tillering, seed color, biomass, plant architecture, yield and 234 activation tagged mutants were selected based on the distinct variations in phenotypic characters. Six mutants that showed distinct variation in photoperiodic response showed almost similar flowering time both under optimal (kharif) and non-optimal (long day) photoperiodic conditions with wild type remained as non-flowered genotype under non-optimal conditions. Further, the flanking sequence analysis of one of the mutants showed integration of the enhancer elements on chromosome 11 flanked by several genes such as Os11g0219000, Os11g0219400, Os11g0218000, Os11g0218100, and Os11g0218500. The bioinformatic analysis of these genes showed that, one of the genes is a bZIP transcription factor which seems to have some role in regulating photoperiodic response. The expression analysis of the flanking genes and their functional validation is under progress.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 334 Session K : Computational and Systems Biology
PK433 (IPPC0110) Characterization of glutaredoxin proteins in monocots Minesh Jethva1*, Saurabh Yadav2 and Hemant R. Kushwaha1 1Synthetic Biology & Biofuels Group, International Centre for Genetic Engineering & Biotech., Aruna Asaf Ali Marg,New Delhi-110067, India 2Department of Biotechnology, HNB Garhwal University, Srinagar Garhwal, Uttarakhand, India *Presenting author: [email protected]
Glutaredoxins (GRXs) are small heat-stable disulfide oxido-reductases and members of the thioredoxin protein family and are involved in maintaining the redox homeostasis in many organisms like bacteria, yeast, and mammalian cells. GRXs exist in entire green lineage and they are classified into dithiol and monothiol by used on the type of active site present. Monothiol and dithiol GRXs contain ‘CGFS’ and ‘CXXC’ as a redox active motif, respectively. The availability of completely sequenced plant genomes allows the comparative study of GRXs distribution across species. In the present analysis, we have studied 19 monocot plant genomes to reveal sequence and domain level diversity within GRX proteins. We have characterized GRX family including chromosome locations of coding genes, protein domain structures, phylogenetic relation and their sub-cellular localization. We have analyzed the phylogenetic distribution of GRXs proteins to reveal evolutionary events like HGT and duplication. This study provides perception of the domain structure conservation of GRXs in monocots. The number of GRXs varies from 39 to 96 in these species but domain structures are relatively conserved.
PK434 (IPPC0199) Hyperspectral reflectance based non-invasive assessment of tomato fruit firmness Vijay Paul1*, Rajeev Kumar1, Rakesh Pandey1, Rabi Narayan Sahoo2, S.K. Jha3, Ram Asrey3, V.K. Gupta2, Atar Singh1 and Tauqueer Ahmad4 1Division of Plant Physiology, 2Division of Agricultural Physics, 3Division of Food Science & PHT, Indian Agricultural Research Institute, New Delhi-110012, India 4Division of Sample Survey, IASR, New Delhi-110012, India *Presenting author: [email protected]
Firmness of tomato fruit is a key parameter to ascertain its quality, transportability, marketing, storage period/shelf-life and preference by consumers. To identify hyperspectral reflectance based indices and to develop models for non-invasive estimation of tomato fruit firmness, freshly harvested and stored tomato fruits of 2 varieties and 2 hybrids representing eight ripening stages were sampled. Reflectance of fruits was taken with spectroradiometer. Reflectance at wavelengths ranging from 350 to 2500 nm was recorded. Fruit firmness (Newton, N) was then determined by texture analyser. Data obtained is analysed for dimension reduction, index identification and model development. Firmness ranged from 11.74 st N (immature fruits) to 1.13 N (over ripe fruits). Out of many indices, two best indices were: R501 (1 derivative) and R521. st Model based on R501 (1 derivative) (= x) is y (firmness, N) = 1260.8 x + 3.3087 with RMSEP = 11.05 N and bias = -0.01. The second model based on R521 (= x) is y = - 5.3768 x2 + 18.209x + 2.9509 having RMSEP = 1.11 N with bias = -0.002. Models were found to be valid irrespective of varieties/hybrids, ripening stages, fresh and stored fruits. Model based on the index R521 is simple (no derivation step) and therefore can also be considered as rapid one. This study will prove useful in developing cost-effective tools. Such tools can then be applied for rapid/automated phenotyping, screening and sorting of tomato fruits based on the firmness at small (lab) or large (industries) scales.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 335 Session K : Computational and Systems Biology
PK435 (IPPC0226) Gravitational biology: significance of the characteristics of statenchyma in dicotyledonous plants A.D. Barnabas1* and P. Dayanandan2 1Post Graduate Department and Research Centre of Botany, The American College, Madurai-625002, Tamil Nadu, India 2Rtd. from the Department of Botany, Madras Christian College, Tambaram, Chennai-600059, India *Presenting author: [email protected]
A specialized group of cells, statocytes, containing starch statoliths (= amyloplasts) appear to be involved in the initial stages of plant gravitropism. The direction of transport of auxin, lateral or radial, can be resolved if we understand the significance of the relative position of the statenchyma (RPS) cells. 141 species of dicotyledonous plants were investigated to characterise the number, dimensions, and relative position of the statoliths and statocytes. Seedlings and young shoots were gravistimulated and the regions of maximal response were sectioned and observed after staining with I2KI. All 141 species possessed statenchyma with sedimentable amyloplasts. The average number of statoliths per statocyte is 10. The statenchyma may be one or more layered and continuous or interrupted. RPS is expressed as an r/R value where r is the radius of the circle defined by statenchyma as the outer limit and R is the radius of the transverse section of the shoot with the epidermis as the outer limit. Very thin shoots have small RPS values, whereas very thick ones have a value of 0.5. In general, the RPS value varies from 0.18 to 0.93. In Peperomia which has scattered vascular bundles, each bundle is surrounded by a layer of statenchyma. In Gomphrena globosa statoliths sediment normally only in young shoots that respond to gravity. The significance of RPS will be discussed in relation to auxin transport and elongation response. Basic research in gravitropism has implications of gravitational biology and space life sciences.
PK436 (IPPC0229) Differential nucleosome occupancy in plant genomes; Are promoter regions significantly depleted of nucleosomes? Amish Kumar* and Gitanjali Yadav National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India *Presenting author: [email protected]
Wrapping of DNA around the nucleosomes core particle is the key to the chromatin packaging as well as for the regulation of gene expression. A comparative study of nucleosome occupancy patterns, at the genome wide level may help to understand chromatin structure and its functional control over the cell. Whole genome nucleosome occupancy maps are available for several organisms and provide sufficient knowledge base to accurately predict the site of occurrence of nucleosomes over a new genome sequence. For example, yeast genome shows differential organization of nucleosomes in the upstream regulatory region of genes, resulting in the existence of distinct nucleosome free regions (NFRs) just before the transcription start site (TSS). This study was designed to test whether such NFRs are present in plant genomes as well, and whether the DNA itself guides differential nucleosome occupancy. Thus in order to understand the role of DNA sequence in nucleosome occupancy, we hypothesized that “There are fewer nucleosomes in promoters compared to other regions of the plant genome”. For this, we analyzed two plant genomes namely, Zea mays (a monocot) and Solanum tuberosum L. (a dicot) to construct genome wide nucleosome prediction maps. Per base nucleosome positioning was compared over the promoter and non-promoter regions of both the genomes. Statistical tests reveal that nucleosomes are indeed depleted in promoters as compared to random, or other genomic regions of plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 336 Session K : Computational and Systems Biology
PK437 (IPPC0268) Bioinformatics of KaiC, a cyanobacterial circadian clock protein Rakesh Yadav* and Namita Singh Department of Bio & Nano Technology, Guru Jambheshwar University of Science & Technology, Hisar-125001, Haryana, India *Presenting author: [email protected]
Circadian clocks are endogenous biological timers that rhythmically control several processes with an around 24-h period. They exist in diverse organisms, but cyanobacteria are the simplest organisms known to possess a clock. The circadian clock protein KaiC, is encoded in the kaiABC operon that regulates circadian rhythms. KaiC have been extensively studied and are widely distributed among Cyanobacteria. In this study, bioinformatics analyses of KaiC proteins were performed in 53 Genera of Cyanobacteria. According to the conserved motifs obtained by MEME and MAST tools, two motifs were common in all proteins. The structural and functional analyses of KaiC proteins were investigated by ProtParam, SOPMA, SignalP 4.1, TMHMM 2.0 and ProDom tools in ExPASy database. The protein sequences were aligned with ClustalW algorithm by MEGA 6.06 software and phylogenetic tree was constructed using the Neighbor-joining (NJ) method. In protein–protein interactions analysis by STRING 9.1 tool, 10 significant protein interaction groups were identified in KaiC. According to the results, there is a high identity among KaiCs of different species in Cyanobacteria, so that they have to be derived from a common ancestor. The approaches and results reported here would be useful to understand biochemical and structural dissection of the molecular basis of a biological clockwork.
PK438 (IPPC0329) Mining Picrorhiza kurroa transcriptomes for transcription factors contributing to picrosides biosynthesis Ira Vashisht*, Tarun Pal and Rajinder Singh Chauhan Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, HP, India *Presenting author: [email protected]
Picrorhiza kurroa is a valuable medicinal herb, recognized for its hepatoprotective, anti-inflammatory, anti-oxidant, immunomodulatory and several other curative properties. Iridoid glycosides such as picroside-I and picroside-II are major ingredients of various pharmaceutical formulations. No information exists on transcriptional regulation of picrosides biosynthesis in P. kurroa. Deciphering complete biosynthetic pathway coupled with expression analysis of genes has revealed that multiple genes are contributing to the biosynthesis of picrosides, thereby, suggesting that common regulatory control affect the biosynthesis of picrosides. Field-grown plants showed significantly higher picroside-I (shoots) and picroside-II (roots) contents compared to in vitro -grown plants. The NGS transcriptomes of P. kurroa tissues with varying amounts of picrosides, were mined for WRKY, MYB, bhlh, bzip, zinc finger, NAC and PHD classes of transcription factors, which have been implicated in regulating secondary metabolites biosynthesis in plants. Among all classes, WRKY exhibited maximum fold variation in field grown shoots (42 folds) and roots (5 folds) as compared to their in vitro counterparts. AP2/ ERF and PHD family of TFs showed negligible expression under in vitro conditions, thereby suggesting their possible involvement in the regulation of picrosides biosynthesis. Functional validation of TFs can provide suitable candidates for enhancing picrosides contents in P. kurroa.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 337 Session K : Computational and Systems Biology
PK439 (IPPC0351) KIXBASE: A KIX domain database and webserver for prediction and analysis Archana Yadav*, Jitendra K. Thakur and Gitanjali Yadav National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
In today’s era of ever-increasing biological data, it is essential to maintain databases that not only integrate and cluster the data as a warehouse but as well help analyze it. Such resources are invaluable tools for researchers as they provide a useful platform to retrieve biological information like sequences, structures, structure classes, pathways etc., thus aiding in the research to a great extent. KIXBASE is a global repository as well as a prediction tool for KIX domains. These domains are present in co-activators and play a central role in the transcription process. KIXBASE has two main parts, a web server and a database. To date, there is no other web resource that provides information on KIX domains. The KIX prediction program detects KIX domains in any organism on the basis of profile hidden markov models developed through alignment of known KIX sequences along with additional filtering criteria for improving accuracy. KIXBASE incorporates the most widely used programs like PSIPRED-3.5 and CLUSTALW2 in the web server for further annotation and quality assessment of the predictions. Users can upload batch entries for protein, genomic or EST sequences in FASTA format and carry out the detection of potential KIX domains, generate the secondary structure for domain of interest and examine conservation with other KIX domains. The backend prediction algorithm is also used for development of the KIX database, which contains 1891 KIX proteins representing 427 organisms spanning metazoans, fungi and plants, comprising the largest online collection of KIX sequences. The prediction algorithm is highly accurate, robust and very efficient in handling multiple input types for KIX prediction.
PK440 (IPPC0360) Non-destructive assessment of tomato colour and ripeness by using reflectance at single wavelength Rajeev Kumar1*, Vijay Paul1, Rakesh Pandey1, Rabi Narayan Sahoo2, Vinod Kumar Gupta2, Anil Dahuja3, Atar Singh1 and Tauqueer Ahmad4 1Division of Plant Physiology, 2Division of Agricultural Physics, 3Division of Biochemistry, IARI, New Delhi-110012, India 4Division of Sample Survey, Indian Agricultural Statistics Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Quality and consumers preference of tomato fruits are primarily determined by its colour and appearance. Rapid and non-destructive methods of assessing tomato colour and ripeness are therefore of immense significance for basic research and applied aspects (processing and value addition). To identify hyperspectral reflectance based indices and developing models for non-destructive estimation of colour and ripeness/maturity of tomato fruits, tomato fruits of different varieties/ hybrids coming from direct harvest or storage were used. Reflectance of individual fruit was taken in a spectrum of 350 to 2500 nm by using a hand-held spectroradiometer. These fruits (immature to over-ripe stages) were then ranked from 0 to 8 based on standard colour chart. Obtained data were subjected to chemometric analysis. All the basic and statistical steps as required for model development were followed. Two models were developed. First model was based on the index R521 (x) i.e., y (colour/ripeness) = -2.456 ln (x) -1.0933. This can accurately predict the colour/ripeness for a difference of > 0.86 (RMSEP) with bias of -0.09. The second model y = 2.582 ln (x) - 0.805 was based on the index R546 (x). This model had RMSEP = 0.89 and bias of 0.12. Models could distinguish different ripening stages of tomato across varieties/hybrids, plant harvested (fresh) and stored (aged) fruits. These findings will prove useful in developing cost- effective tools for rapid sorting and screening of huge collection of harvested tomato fruits as desired for monitoring and by the processing and pharmaceutical industries.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 338 Session K : Computational and Systems Biology
PK441 (IPPC0400) Evolutionary significance of the CBM49-modulated Type C GH9-cellulase activity in plants Siddhartha Kundu* and Rita Sharma School of Computational and Integrative Sciences, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Members of the GH9 (endo-1, 4-glucanase, EC 3.2.1.4) clan have been ascribed roles in the generation of biofuels, as initiators and facilitators of plant development (elongation, abscission), and as mediators of abiotic stress response pathways. Catalytically, these enzymes cleave the β (1-4) glycosidic linkage between pairs of β-D-glucopyranose residues from the non-reducing ends of cellulose in regions best described as amorphous. The Type C subfamily possesses a unique carbohydrate binding domain (CBM49), which is cleaved during processing of the primary transcript. Evidence suggests that the delineation from the classical Type B domain is fuzzy, thereby, raising the possibility of a truncated contribution to the active site of Type C enzymes. To comprehend the significance of the CBM49-modifying Type C GH9- cellulase activity, an evolutionary profile spanning several millennia was constructed. Previous work has suggested that the division into sub-families (Types –A, -B, -C) is entirely anagenetic (intra-lineage). Here, we generate HMM-based datasets of putative sequences spread over several taxa. Our sequence-centric approach is comparatively coarse-grained, and focuses on patterns of bases/ amino acids. The significance of these intra-sequence clusters is a relatively novel annotation schema for GH9-orthologs in existing genomic data. These, when subjected to evolutionary analysis, could offer insights into speciation-level events as well as posit functional roles for the phyletic subgrouping of A-, B- and C- members. The upgradation of Type C endo-glucanases to include micro-crystalline cellulose as a substrate, could, possibly simplify the search for the perfect biofuel.
PK442 (IPPC0471) A smart soft tool to integrate climate model and grid based weather data Gayathri Chamanthula*, B. Sailaja, S.R. Voleti and D. Subrahmanyam ICAR-Directorate of Rice Research, Rajendranagar, Hyderabad-500030, AP, India *Presenting author: [email protected]
Rice crop is climatically the most adaptable cereal. Management of rice crop is difficult as it is grown over a large spatial domain and wide range of landscape types. The projected changes in temperature will further complicate the management of rice crop. Forecasting grid based future temperatures and identifying vulnerable temperature grids to raise rice yields at regional level in varied rice growing environments. With this background, ClimGen climate model has been chosen to integrate with the grid based temperature Map. Grid wise temperature data was collected from IMD for 15 years and total 346 grid points were there for temperature. A software program was developed to convert individual text files supplied by IMD into one single database and computing day and night temperatures. These files were converted to geo database files using Arcatalog and thessien polygons were generated for each point using spatial interpolation method of ArcGIS package. An interface has been created to integrate this grid based weather map with climate model. P-mapper an open source software is used for this program. ClimGen model was executed for each grid and generated long term series up to 2050. Weekly maps were generated for mean temperature and difference in day and night temperature. Vulnerable temperature zones were identified at the time of flowering (February and March) during rabi season for the years 2030 and 2050. This software is user friendly and further integration with any kind of maps (point/polygon wise) and models is possible with this interface.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 339 Session K : Computational and Systems Biology
PK443 (IPPC0607) Integrative co-expression network analysis to infer the immune response in rice during the invasion of rice blast fungus Preeti Arya* and Vishal Acharya CSIR-Institute of Himalayan Bioresoruce Technology, Palampur-1760061 (H.P.), India *Presenting author: [email protected]
Rice is an important staple cereal crop grown world-wide and is among the highly consumed food commodity. The various diseases in rice cost for an average of 37% reduction in the yield globally. Rice blast fungus (Magnaporthe) is the most common phyto-pathogenic fungi responsible for substantial loss in production. The mechanism by which pathogens infect its host is the most important and still unresolved aspect for many host-pathogen systems. Various studies pertain to decipher the mechanism of host-pathogen interaction; however, in vivo/in vitro methods are laborious, time consuming and are not high-throughput. In silico approach may be the best possible alternative to mine the useful information regarding pathogenesis using data generated through microarray and highly advanced RNA sequencing. In the present study, the publicly available transcriptome data for rice and Magnaporthe was retrieved from NCBI GEO and used to identify the expression profiles of differentially expressed gene (DEGs) sets. The co-expression network was constructed by means of the Pearson correlation coefficient (PCC) between expression profiles of DEG(s) using weighted correlation network analysis (WGCNA) that were further employed for finding the highly correlated gene modules, the biological targets of interest. Moreover, the protein-protein interaction and interlog mapping was used to validate interacted gene pairs obtained from the co-expression networks. The resulted interactions will be further evaluated statistically using computational strategies and also on basis of experimentally known pathogen-host protein interactions. This rice- Magnaporthe interactome hence will provide valuable insights to understand the mechanism of pathogenesis by targeting hubs responsible for the disease development.
PK444 (IPPC0677) Tissue specificity gene expression database Ambarish Kumar*, Vijya Brahma and Andrew Lymm Jawaharlal Nehru University, New Delhi-110067, India *Present author: [email protected]
The RNA-Seq is the next generation sequencing technology to get a snapshot of RNA present and quantify (transcriptome expression profile) from a genome at a given moment of time. The expression data from the whole transcriptome can provide important information about mRNA levels revealing stage specific and localized gene expression. Although many tissue specific expression databases are available (TIGER, TiSGeD, Very Gene etc.) but till date no work has been reported with respect to the plant transcriptome expression, ensembled, based on the tissues (leaves, flower, fruit, roots etc.), stages of development and localization. The database specific to plant tissue specificity can shed light upon the disease biology, applied medical research, pharmaceutical drug development , biomarker discovery in the field of translational medicine, proteome classification on the basis of tissue specificity, nutritional development as well as increase in the productivity of certain crops. These information from the known repository will be collected, classified and its liaison with the assembled and annotated transcriptome will be surmised. The curated dataset for clade of Viridiplantae encompassing all the green plants was retrieved from UNIPROTKB (http://www.uniprot.org/). A total of 3353453 fasta sequences and their corresponding tissue specificity, stages of development, localization and pathways data was used for creating the database. We propose to integrate onto Galaxy based platform, wherein the RNA-seq data will be assembled (de novo/reference), analyzed for differential expression, followed by the annotations using BLAST algorithm and the PTSDb will be linked to the annotations.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 340 Session K : Computational and Systems Biology
PK445 (IPPC0821) Molecular interaction pattern of sugarcane streak mosaic virus encoded micro RNA, SCSMV_miR16 in sugarcane - A In silico approach Prabu Gajjeraman* and Yasir Umar Pattiwala Plant Functional Genomics Unit, Department of Biotechnology, Karpagam Academy of Higher Education, Coimbatore-641021, India *Presenting author: [email protected]
Sugarcane Streak Mosaic Virus (SCSMV) is a distinct member of Poacevirus in Potyviridae family which has a single stranded positive-sense RNA genome causing mosaic disease in sugarcane. Among the various regulatory mechanisms, gene silencing was found to play a major regulatory role during disease progression. Micro RNAs (miRNAs) are recently identified small RNA molecules (~24 nt bp in length) which are involved in regulation of host plant gene expression and vice versa in association with argonaute (AGO) proteins via RNA silencing. To study the gene regulatory pattern, we analyzed the most preferential SCSMV_ miR16 – target mRNA – argonate protein interaction at molecular level by in silico approach. The hydrogen bonding and other interactions were analyzed and discussed. These finding considerably broaden future investigation of the AGO-assisted miRNA based gene regulation during SCSMV pathogenesis in sugarcane.
PK446 (IPPC0873) Morpho-physiological characterization of cashew accessions in west coast region of Karnataka Babli Mog1*, P.L. Saroj1, T.G. Prasad2 and Vishwanathan Chinnusamy3 1Plant Physiology, Directorate of Cashew Research, Puttur-574202, Karnataka, India 2University of Agricultural Sciences, Bangalore, Karnataka, India 3Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
Cashew (Anacardium occidentale L.) is an important nut crop of social and economic importance for India. The low productivity of raw cashew nut is the major constraint. As detailed physiological characterization of cashew accessions is the key step for yield improvement, this study was carried out to analyze genotypic variation in yield and yield components of 13 cashew accessions in twelve year old plantation. Significant differences in leaf traits were observed among cashew accessions. Maximum leaf ratio was observed in VRI-3 while Bhaskara showed maximum leaf length and width. Maximum of 28 main veins were recorded in Ullal-3. Highest leaf area and specific leaf weight (SLW) were recorded in VTH-30/ 4(112.67 cm2) and Vengurla-3 (17.43 mg/cm2), respectively. The number of leaves per lateral shoot was more in VTH- 174, Bhaskara, VRI-3 and VTH-30/4. Ullal-3 exhibited higher RWC and lower rate of excised leaf water loss (10.8%) followed by Bhaskara. The contents of Chl a, Chl b, total chlorophyll and carotenoids were highest in Bhaskara. VTH-174 showed highest auxillary bud differentiation into reproductive structures followed by Bhaskara. Significant variation for nut development observed among accessions. VTH-174 took least time for apple development (56 days) as well as final harvest (60 days). VTH-30/4 recorded highest apple with nut weight, apple and nut fresh weight. Brix value of apple was recorded maximum in VTH-174 indicating it’s sweetness with less tannin content. The relationship between component physiological traits and yield will be presented.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 341 Session K : Computational and Systems Biology
PK447 (IPPC0900) Elucidating the alkaline tolerance mechanism of Arthrospira platensis: An In-silico study Monikam Jangir* and Jyoti kashyap BITS Pilani, Hyderabad Campus, Hyderabad, India *Presenting author: [email protected]
Alkalophiles, a class of extremophiles are capable of sustaining a high alkaline environment (~pH 11) but their exact mechanism is unknown. The alkalotolerant mechanism could be extrapolated to improve productivity of higher plants facing stressful environment. Out of various suggested hypothesis, Na+/H+ antiporters mediated regulation is the widely accepted one which corroborates their role in pH homeostasis. Sequence and structural analysis was performed to understand precise role of antiporters. We analyze antiporter sequences of alkalophilic Arthrospira platensis to understand the underlying mechanism while the antiporters of alkalotolerant Synechocystis spp are considered as a control. Difference in copy number and the presence of various functional domain(s) indicates the differential regulation of pH by these antiporters. Greater than 50% identity was obtained between 3 of the antiporters sequences of the two organisms. NhaP, KefB were the domains present in both organisms along with a unique UspA domain (Universal stress Protein A) only in Arthrospira platensis. Transmembrane regions were also deduced using online servers in which Arthrospira platensis showed presence of 11-12 helices while Synechocystis spp showed 9-12 helices. This study pointed out possible role of antiporters on alkali tolerance.
PK448 (IPPC0947) The sorghum gene expression database Muhammad Ali Imam1, Vinay Singh2*, Manoj K. Sharma2 and Rita Sharma1 1School of Computational & Integrative Sciences, 2School of Biotechnology, Jawaharlal Nehru University, New Delhi-110067, India *Presenting author: [email protected]
Sorghum bicolor, recognized as an alternate feedstock for biofuel production in many parts of the world, is a multipurpose crop used for food, feed, fodder as well as fibre. Due to relatively small genome, availability of whole genome sequence and low level of gene duplication, it is also being used as a model for understanding the genetic mechanism underlying C4 photosynthesis in grasses.Till date, both microarrays and RNA sequencing technologies have been used to elucidate transcriptional dynamics during different developmental stages of sorghum as well as in response to stress. To facilitate functional genomics studies and development of improved varieties, we are developing a freely available user-friendly community resource using publicly-available sorghum expression data. Both microarrays and RNA sequencing-based transcriptomic data has been downloaded from NCBI-Gene Expression Omnibus and normalized using a standard pipeline. The data can be retrieved using microarray elements IDs or gene accession numbers. Our web interface will allow visualization and retrieval of gene expression profiling data via heat map and bar graphs in selected experiments/samples. Additional tools for coexpression, differential expression and pathway analysis with the genes of interest will also be provided. Therefore, this database will serve as a useful reference for elucidating gene expression and functions of sorghum genes.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 342 Session K : Computational and Systems Biology
PK449 (IPPC0949) An approach for high throughput phenotyping of tomato genotypes using Plant Phenomics Platform R.H. Laxman*, P. Hemamalini, R.M. Bhatt and A.T. Sadashiva ICAR-Indian Institute of Horticultural Research, Bengaluru, Karnataka *Presenting author: [email protected]
Physiologists constantly strive to phenotype plants for identification of desirable traits for their utility in crop improvement. The plants’ performance is quantified through measurement of various parameters under challenging conditions both in field and controlled conditions. Both, destructive and non-destructive approaches are adopted in this endeavour. Since, the detailed traditional methods of plant phenotyping are time consuming, laborious and involve human errors; the focus is being shifted towards non-invasive imaging and image analysis which are high throughput in nature. The high throughput phenotyping approaches being non-destructive enable imaging of the same plant at different developmental phases. Considering these inherent advantages, studies were conducted in the Plant Phenomics facility with the objective to evaluate the tomato cultivars, hybrids and germplasm lines at various developmental stages. The container grown plants were imaged and growth performance was quantified. The imaged plants were sampled for measurement of the actual biomass. The study showed significant relationship between actual biomass and digital biomass and through imaging technique it was possible to quantify biomass across the genotypes and also at different growth stages. The utility of Plant phenomics facility for high throughput evaluation and phenotyping of tomato genotypes are discussed in this paper.
PK450 (IPPC0959) Quantitative monitoring of sucrose dynamics for phenotyping of soil moisture-deficit stress tolerance in rice using hyper-spectral reflectance data R.N. Sahoo1, Bappa Das1, G. Krishna1, Puja Rai2, Rahul Tiwari2*, R. Verma2, C. Viswanathan2, S.K. Dash4, P. Swain4 and P.C. Nautiyal3 1Division of Agricultural Physics, 2Division of Plant Physiology, 3Division of Seed Science and Technology, Indian Agricultural Research Institute, New Delhi-110012, India 4ICARI-Central Rice Research Institute, Cuttack, Odisha, India *Presenting author: [email protected]
The capacity of rice to store sucrose in the stem is regarded as a promising trait to buffer yield in environments with limited water availability. Measurement of volume and rate of flow of sucrose to the panicle during grain filling under water-deficit condition could be one of the reliable tools for measuring soil moisture-deficit stress tolerance than the currently used tools. Keeping this in view, a study was conducted for non invasive estimation of sucrose in rice stem using hyper-spectral reflectance data collected in the spectral range of 350 to 2500 nm for different rice genotypes grown under different soil moisture deficit stress levels. Plant samples were collected synchronizing with spectral measurements for estimation of sucrose. Spectral data analysis was done to identify suitable spectral indices and models for sucrose estimation. Two spectral indices, viz., ratio of sucrose index (RSI) and Normalized Difference Sucrose Index (NDSI) in NIR range were proposed with reliable accuracy of sucrose estimation. Different multivariate spectral models such as Partial least squares regression (PLSR) and multiple regression model (MLR), Multivariate adaptive regression splines (MARS), Support Vector Machine (SVM) and random forest (RF) were also evaluated. MARS was found to predict the sugar content with better accuracy than others, whereas the prediction accuracy of RF was the poorest. The proposed non invasive in situ method was found to be alternate fast reliable method to labour intensive laboratory methods and a high throughput field applicable phenotyping technique for water deficit stress studies and rice breeding programs.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 343 Session K : Computational and Systems Biology
PK451 (IPPC0984) Application of thermal imaging to monitor the crop stress condition P. Krishnan*, Kaushik Banerjee and Victor Banerjee Division of Agricultural Physics, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected] Monitoring the crop stress condition is an imminent requirement for abiotic stress characterisation. In this study, non- destructive and non-invasive characterisation of crop canopy status of wheat (Triticum aestivum) grown under moisture and temperature stress, was analysed using thermal imaging. When wheat crop is under abiotic stress there is a decrease in latent heat flux at the crop canopy surface, a complementary increase of sensible heat occurs and creates a larger temperature difference between crop canopy and air. Recent thermal imagers can instantly detect minor differences in crop canopy temperatures which correlate to changes in transpiration, thus revealing differences in crop stress condition. The major interest in the application of thermal sensing of field crop was the development of indices of crop stress as guidance for crop management. In this study a crop canopy stress index (CCSI) was derived from the image analysis of the thermogram of the crop. A significant correlation was observed between the CCSI derived from the thermogram with the plant stress indicators like stomatal conductance, Relative Water Content, Water Potential and Leaf Area Index. In addition a high correlation was found between yield and CCSI at different growth stages indicate that thermography can characterise the crop stress status in wheat. These indices are mostly presumed to reveal variations in stomatal opening and evaporation rate as cellular water becomes the stress regulating factor. Thus thermal imaging as observed in this study can be an efficient tool to characterise the crop responses to abiotic stress conditions.
PK452 (IPPC0989) Comparative identification of microRNAs and their potential targets between Capsicum chinense and Capsicum frutescens Mukesh Nitin1*, Rajnish Kumar1, Onkar Nath2, Nirala Ramchiary1, Sarpras M1, Rashmi Gaur1, Vineet Sharma1, Sushil Chhapeker1, Toshi Mishra3, Vijaya Bramha2 and Andrew Lynn2 1School of Life Sciences, 2School of Computational and Integrative Sciences, Jawaharlal Nehru University, New Delhi-110067, India 3Department of Bioinformatics, BHU, Varanasi, India *Presenting author: [email protected] MicroRNAs (miRNAs) are small non-coding RNAs which regulate gene expression involved in various developmental processes of plants and animals. Although many miRNAs and their potential target genes have been identified in various crop plants, there is very limited information in Capsicum species. Therefore, here in this study, we used transcriptome data of Capsicum frutescens and C. chinense to identify miRNAs and their potential target genes. The transcriptome data were generated from leaf, flower and three fruit stages of each Capsicum species using Illumina High Seq2000 sequencing machine. Using matured miRNAs of already reported plant miRNAs from Plant Non coding RNAs database (PNRD) and miRbase against 184975 C. chinense and 179780 C. frutescens transcripts, we identified a total of 747 and 708 potential miRNA transcripts in C. chinense and C. frutescens, respectively. Further structure folding and minimum folding free energy (MFE) analysis with VieenaRNAfold programe using standard criteria finally gave a total of 127 potential miRNAs in C. chinense and 146 miRNAs in C. futescens, of which 9 and 18 miRNAs showed differential expression in different tissues of C. chinense and C. frutescens respectively. Out of predicted miRNAs in both the species, majority of miRNAs were conserved but few are found species specific. The prediction of potential targets for differentially expressed miRNAs using PsRNAtarget programe identified a total of 218 and 360 mRNA target genes in C. chinense and C. frutescens, respectively, which are involved in various cellular functions. Few miRNA families showing differential expression in C. chinense and C. frutescens, were experimentally validated using qRT-PCR. Development of miRNAs based molecular markers is being in progress which could be used in Capsicum breeding programe.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 344 Session L : Climate Change and Agriculture
PL453 (IPPC0073) Impact of reduced PAR in determinate, semi-determinate and indeterminate soybean (Glycine max L.) genotypes under climate change scenario Kiran Bhagat*, Santanu Bal, Yogeshwar Singh, Ratnakumar Pasala, Sunayan Saha and P.S. Minhas National Institute of Abiotic Stress Management (NIAM), ICAR, Malegaon, Baramati-413115, Pune, Maharashtra *Presenting author: [email protected]
Soybean is an important crop, which is photosensitive in nature and hence will be highly affected by the increasing intensity of atmospheric brown clouds, commonly known as “surface dimming”, and moisture stress condition, which may prevail in the near future. Under present climatic condition, the average incoming PAR during morning hours is around 1300 during kharif season, and it is predicted to decrease by 15-20% in near future. Three different structures were tested to standardize the uniformity of PAR interception within the structures. Perusal of data clearly revealed that as there is homogeneity in every aspect in semi-circular dome shape structure as compared to rectangular and octagonal dome structures, and therefore, semi-circular dome shape structure was considered most appropriate for conducting such type of research work. Light saturation point (LSP) calibration revealed that determinate soybean genotype showed LSP near to 800 μmol m-2 s-1 PAR, with photosynthetic rate of 20.77 μmol m-2 s-1, whereas semi-determinate genotype showed LSP near to 1100 μmol m-2 s-1 PAR, with photosynthetic rate of 21.95 μmol m-2 s-1, and indeterminate genotype showed LSP near to 1000 μmol m-2 s-1 PAR, with photosynthetic rate of 28.88 μmol m-2 s-1. Determinate, semi-determinate and indeterminate soybean genotypes tested under control and in semi-circular dome shape structure by using white shade net with 15-25% PAR reduction factor, revealed that yield of indeterminate soybean genotype was higher than determinate and semi-determinate types under reduced PAR. Hence, indeterminate soybean genotypes may perform better under reduced PAR condition under future climate change scenario.
PL454 (IPPC0148)
Effect of elevated CO2 and temperature on wheat and Phalaris minor Kamlesh Paraste, P.P. Choudhury, M. Rathore, R. Singh, C. Sarathambal, N. Tripathi and Bhumesh Kumar* ICAR-Directorate of Weed Research, Jabalpur-482004, India *Presenting author: [email protected]
Effect of elevated CO2 and elevated temperature on wheat and Phalaris minor (an important weed of wheat crop) was studied in open top chambers (OTCs). Wheat (cv. GW-273) was sown following recommended practices and weed population was maintained 15 plants/m2. Plants of both the species were exposed to different treatments [viz. ambient, o elevated CO2 (550 ± 50 ppm), elevated temperature (ambient + 3.0 ± 0.5 C) and elevated CO2 + elevated temperature] from emergence to physiological maturity. Enrichment of atmospheric CO2 had a positive effect on overall growth of wheat as well as P. minor, however, elevated temperature alone or in combination of elevated CO2 had adverse effect on growth and development of both the species and more severely of wheat as compared to P. minor. Rate of photosynthesis increased in both the species when plants were exposed to elevated CO2, however, a decrease in photosynthesis was observed at elevated temperature alone or in combination with elevated CO2.Species-specific as well as treatment- specific differential expression of genes involved in photosynthesis and defence pathways was evident. As compared to ambient conditions, advancement of panicle emergence was observed in both the species when exposed to elevated temperature (by 15 days), elevated CO2 (by 7 days), and combination of elevated temperature and CO2 (by 10 days).
Efficacy of sulfosulfuron against P. minor was considerably reduced at elevated CO2 and elevated temperature indicating adverse effects on weed management. Results suggest that climate change may alter crop-weed interaction in favour of weeds.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 345 Session L : Climate Change and Agriculture
PL455 (IPPC0164) Lodging: A new face of climate change devastating agricultural productivity Rinki Khobra*, H.M. Mamrutha, Raj Kumar Gupta and Indu Sharma Division of Crop Improvement, Indian Institute of Wheat and Barley Research, Karnal-132001, Haryana, India *Presenting author: [email protected]
Lodging is a complex phenomenon that is influenced by interaction of crop plant, rain, wind, soil and result into permanent displacement of plant stem from its normal position. Introduction of dwarfing gene, has increased lodging resistance in Indian wheat varieties and hence lodging was not considered as a major problem till date. But, due to untimely rainfall in the major wheat growing area (North West Plain Zone) during, 2014-15 crop season, significant (5-7%) crop lodging has been reported. Hence, it is need of hour to explore the effect of lodging for developing lodging resistant wheat varieties for secured agriculture and food security in India. As an initial attempt to analyze the post harvest effect on wheat crop lodging, some of the morphological, physiological and quality traits were studied in the grains of lodged and non-lodged genotypes. The grain samples were taken from IIWBR field trial having 25 genotypes with two replications in two conditions during 2014-15 crop seasons. The results of initial screening showed a significant reduction in germination percentage, shoot length and protein content in lodged genotypes but increase in their test weight. Significant visual differences in grain color, size and luster were also observed. However these results are very preliminary and more study is required to understand the negative effect of lodging on wheat grains and to develop the non-lodging wheat ideotype.
PL456 (IPPC0313) Effect of water management practices during post-vegetative stage under system of rice intensification (SRI) Amod Kumar Thakur*, K.G. Mandal and S. Raychaudhuri ICAR-Indian Institute of Water Management, Bhubaneswar-751023, Odisha, India *Presenting author: [email protected]
The system of rice intensification (SRI) has been promoted for enhancing yield using less water. SRI recommendation for water management is to apply a minimum of water during the vegetative stage, and then to maintain a thin layer of water during post-vegetative stage. Moreover, it is not clear about optimum water management during post-vegetative phase of rice crop for the best results. The present study was carried out to investigate the impact of continuous vs. alternate flooding of paddy fields during post-vegetative stage on crop phenotype, physiology and performance under two crop management systems, namely, SRI and conventional transplanting system (CTS). During vegetative stage of crop growth, CTS plots were kept flooded and in SRI plots 3-DAD (days after disappearance of ponded water) irrigation method were followed. Post-vegetative stage, four water management treatments, viz., continuous flooding (CF), 1-DAD, 3-DAD and 5-DAD were imposed both in CTS and SRI. Rice grown with SRI practices showed significant improvements in the plants’ morphology-plant height, tillering, leaves number, and expanded root systems. These were accompanied by improvements in physiological functions like xylem exudation rate, canopy light interception, leaf chlorophyll content, light utilization, and photosynthetic rates during flowering. These factors were responsible for increased grain yield as compared with CTS. Significantly higher water productivity was found under SRI (6.3 kg/ha-mm) than CTS (3.3 kg/ha-mm). Under CTS, the highest water productivity was obtained with the 1-DAD treatment, while under SRI, it was achieved in the 3-DAD treatment.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 346 Session L : Climate Change and Agriculture
PL457 (IPPC0339)
Effect of elevated CO2 level on fate of azoxystrobin fungicide and soil microbial parameters in rice soil Suman Manna1, V.P. Singh2 and Neera Singh1* 1Division of Agricultural Chemicals, 2Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi-110012, India *Presenting author: [email protected]
A major indirect response to increased CO2 levels in the atmosphere is the greater belowground C allocation through root exudation, which is likely to lead to change in size and activity of soil microflora and would have direct effect by increasing temperature. Both may affect the degradation and persistence of pesticides in soil used for crop protection. We report the effect of elevated CO2 (580±20 ppm) on the degradation of azoxystrobin [methyl (E)-2-{2-(6-(2-cyanophenoxy) pyrimidin- 4-yloxy) phenyl}-3-methoxyacrylate] and soil microbial activity in rice planted soil in open top chambers (OTC). Results indicated that there was not much difference on the persistence of azoxystrobin in rice planted soil maintained under different CO2 levels. The half life values for the azoxystrobin in rice soils were 20.3 days in control (rice grown at ambient
CO2 outdoor), 19.3 days in rice grown under ambient CO2 atmosphere in OTC and 17.5 days in rice grown under elevated -1 CO2 atmosphere (580±20 μmol mol ) in OTC. Azoxystrobin acid was recovered as the only metabolite of azoxystrobin, but it did not accumulate in the soil/water and was further metabolized. Elevated CO2 enhanced soil microbial biomass carbon (MBC) and alkaline phosphatase activity in soil. Compared with rice grown at ambient CO2 (both outdoors and in
OTC), the soil MBC at elevated CO2 increased by two fold. Elevated CO2 did not affect dehydrogenase, fluorescein diacetate (FDA) and acid phosphatase activities. Azoxystrobin application to soils, both under ambient and elevated CO2 levels inhibited alkaline phosphates activity, while no effect was observed on activity of other enzymes.
PL458 (IPPC0355) Physiological and biochemical plasticity of Lepidium latifolium L. suggests it as sleeper weed in western Himalayas under climate change scenario Dhiraj Vyas*, Tarandeep Kaur and Hilal Bhat Biodiversity and Applied Botany, Indian Institute of Integrative Medicine (CSIR), Canal Road, Jammu-180001, J&K, India *Presenting author: [email protected]
Unlike the aggressive invasive character of Lepidium latifolium in riparian plains of North America, its growth is restricted in Ladakh Himalayas. In order to understand its future spread under climate change scenario in ecologically sensitive region of Western Himalayas, native populations of Lepidium latifolium in different microclimates were studied for physiological and biochemical plasticity. Different populations from sparsly populated site (SPS; 2807 m asl; 34.54 N, 76.13 E), moderately populated site (MPS; 4139 m asl; 33.20 N, 78.64 E) and densly populated site (DPS; 3164 m asl; 34.15 N, 77.57 E) were studied for their fluorescence, photosynthetic and biochemical characteristics. Results showed that the DPS had higher photosynthetic accumulation and transpiration rate despite lower VpdL and higher relative humidity. This suggests the regulation of its leaf temperature by evaporative cooling. Apart from its nutritive value, higher amounts of sinigrin obtained in DPS might play an important role in water transport under environmental stress. The quantum efficiency of PSII photochemistry (Fv/Fm, NPQ, PSII) and light curve at various PPFD’s suggests better light harvesting potential and light compensation point at DPS than the other two sites. Concomitantly, plants at SPS had significantly higher lipid peroxidation, suggesting a stressful environment, and higher induction of antioxidative enzymes. High light intensities at MPS are managed by specialized contrive of carotenoid pigments and PsBs gene product. It is suggested that L. latifolium is present as ‘sleeper weed’ that has inherent biochemical plasticity involving multiple processes and its potential spread is linked to site-specific micro-environment.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 347 Session L : Climate Change and Agriculture
PL459 (IPPC0386) Physiological basis for development of climate resilient cultivars R.K. Bhatt*, V.K. Manga, R. Sharma, A. Saxena and Uday Burman Central Arid Zone Research Institute, Jodhpur-342003, Rajasthan, India *Presenting author: [email protected]
Climate change is experienced by increase in temperature and extreme weather events. It is projected that its impact will be more significant in the coming years. Research is required to enhance crop resilience and adaptation to climate change for sustainable production, food and nutrition security. This may be achieved by developing varieties with in-built developmental plasticity to tolerate high temperature and soil moisture stress particularly at flowering and grain filling stage, resulting in higher productivity. Screening under varying environments in pearl millet showed that some populations (CZP2K9, CZP 923, CZP2K3, CZP9802) and hybrids (CZH226, CZH227, CZH225, ICMH 356) were promising climate resilient genotypes based on yield. CZP2K9, CZP2K11, CZH227, CZH226, GHB538 reflected less than 10% variation in leaf water content and membrane stability index. CZP2K9 also recorded higher grain yield with higher water use efficiency over the environments differing in total rainfall, temperature and number of rainy days. In clusterbean genotypes HG884, RGC1066, RGR-07, RGC1031, CAZG11-1 and RGC936 were selected as promising on the basis of grain yield and less than 10% variation in leaf water content. In terms of membrane stability index, however, only HGS-03-52 and RGC1031 recorded less than 10% variation. Seed yield significantly correlated with dry weight of root and could be considered as one of the parameters for screening the germplasm for hot arid environment. Thus, genotypes exhibiting more resilience with respect to yield and physiological parameters to variable climatic conditions could form the backbone of future breeding programs in both pearl millet and clusterbean.
PL460 (IPPC0716) Phenotyping guar germplasm suitable for rainfed ecosystem of Tamil Nadu N. Sritharan1* and A. Manivannan2 1Department of Crop Physiology, 2Central Institute for Cotton Research, Tamil Nadu Agricultural Univ., Coimbatore-641003, India *Presenting author: [email protected]
Guar (Cyamopsis tetragonoloba L. Taub.) production is greatly affected by climatic factors (rainfall and temperature), soil factors and cultural practices. Although guar is grown under wide range of annual precipitation, the amount and distribution of rainfall is the limiting factor for guar production under semiarid regions. In this light, the objectives were formulated to study the performance of guar germplasm based on morpho-physiological traits as well as to assess the yield attributes under rainfed ecosystem. A field experiment was conducted at Agricultural Research Station, Kovilpatti during the year 2014 rabi season, and a total of 42 guar genotypes (including vegetable, fodder and gummy types) were screened under rainfed condition. Plant morphological characters, chlorophyll index, total dry matter production (TDMP), yield and yield attributes were recorded. Soil moisture status at different depth was also measured. The fodder type, HFG 119 recorded higher grain yield (824 kg/ha). In the case of gummy types, GAU513 recorded superior seed yield (818 kg/ha) which was on par with HGS3-2, RGC 1003 and CAZG10. Among the vegetable types, T local recorded greater seed yield (607 kg/ ha). In correlation studies, TDMP, leaf area, leaf area index and number of seeds per pod were positively correlated with grain yield.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 348 Session L : Climate Change and Agriculture
PL461 (IPPC0800) Oryza2000 model for effective rice crop management under irrigated ecosystem B. Sailaja*, S.R. Voleti and D. Subrahmanyam ICAR-Indian Institute of Rice Research, Rajendranagar, Hyderabad, India *Presenting author: [email protected]
Biophysical models predict the behaviour of the land use system in physical terms such as crop yields, environmental effects, and effect on management. Oryza2000 model is an eco-physiological crop model to simulate the growth and development of rice crop in situations of potential production, water limitations, and nitrogen limitations. This paper reports on the calibration and validation of the model under irrigated ecosystem using independent data sets of wet season rice crop. Data are originated from the nitrogen and water limited experiments conducted by the physiology department of IIRR, Hyderabad 2006. The calibration of the model was performed by deriving the crop growth parameters for long (BPT 5204), mid (Ajaya) and short (Rasi) duration varieties suitable to the local climatic conditions. Oryza2000 model was able to simulate the site-specific rice production with good performance at high nitrogen level as simulated values were very close to observed values. On the other, overestimation of leaf area index (LAI) at all nitrogen levels was consistent indicating a need in this area for further improvement of the model. Model performance was also assessed by statistical analysis (R2, D-index and NOF). D-index is *0.8–0.9 for two varieties at 3 nitrogen application levels for Ajaya and BPT varieties. The results indicated applicability of model Oryza2000 can be extended to rice genotypes varying in their growth periods and situations like fertilizer, water limited conditions.
PL462 (IPPC0972) Morpho-Physiological efficiency of Soybean (Glycine max L.) cultivars under acid soils of mid-altitudes of Eastern Himalayas Krishnappa Rangappa1*, Azad Singh Panwar2, Lotika Kalita2, Meghna Haloi1, Hadienlarisa Syiemlieh2, Savita3, Anjan Kumar Sarma4, Nishant Anandrao Deshmukh5 Anup Das3 and Shishom Vanao Ngachan6 1Plant Physiology Lab, 2Integrated Farming System Section, 3Agronomy Section, Division of Crop Production, ICAR Research complex for NEH Region, Umiam-793103, Meghalaya, India 4Division of Natural Resource Management, 5Division of Horticulture, 6Director, ICAR Research complex for NEH Region, Umiam-793103, Meghalaya, India *Presenting author: [email protected]
Acid soils predominantly present in EHR (80% total geographical area) limit the productivity of most of agricultural and horticultural crops to great extent. Increased concentrations of Al, Fe, Mn, Cu and reduced availability of phosphorus in these acid soils significantly influence physiological, biochemical and molecular plant response. In this connection, present investigation aimed at characterising the morpho- physiological response of 14 soybean cultivars under severly affected acid soils of upland conditions in mid-altitudes of Eastern Himalaya. Results show that, soybean cultivars exhibited significantly varied soil acidity tolerant physiological adaptations in terms of differential root growth traits especially in rhizosphere acidification, root architecture and R/S ratio, altered leaf characteristics viz., leaf pigmentation, leaf CMS, stomatal size and density), changed total plant biomass and nutrient accumulation in root and leaves which in turn influence the yield and yield attributes of the soybean plant in later stages. This altered pattern of morpho-physiological response of different soybean cultivars might have dependent on the extent of soil nutrient regimes under acid soil. This physiological characterisation and identification of efficient soybean cultivars can be further used in breeding program for developing suitable soybean genotypes tolerant to acid soils of Eastern Himalaya.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 349 Session M : Reproductive Biology
PM463 (IPPC0294) OsMADS29: A newly evolved regulator for an ancient metabolic pathway Vibha Verma1*, Meenu Kapoor2 and Sanjay Kapoor1 1Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India 2University School of Biotechnology, Guru Gobind Singh Indraprastha University, New Delhi-110075, India *Presenting author: [email protected]
OsMADS29 is a cereal seed-specific transcription factor that has been shown to affect hormone homeostasis, causing several-fold increase in the levels of active cytokinin (trans-zeatin) and a marked decrease in the levels of auxins. In order to investigate if (a) there existed a conserved biochemical and/or regulatory pathway for cytokinin metabolism in diverse taxonomic groups in the plant kingdom and (b) that rice MADS29 could exploit it for the purpose of starch accumulation, we have generated tobacco BY-2 and Physcomitrella patens transgenic lines expressing rice MADS29 under maize ubiquitin and 35S CaMV promoters. We had earlier shown that the expression of OsMADS29 could mimic the effects of exogenously added cytokinins in the heterologous, tobacco BY-2 cell line, system. Our recent work indicates that the OsMADS29 expression in BY-2 cell lines causes early and enhanced differentiation of proplastids into amyloplasts. This differentiation is associated with positive modulation of several genes involved in plastid biogenesis and starch biosynthesis. In moss also, OsMADS29 expression results in several developmental and morphological abnormalities that are suggestive of excess of cytokinins in the system. These results are suggestive of the existence of a cytokinin metabolism regulatory system that has largely remained conserved during the course of evolution from lower plants to present day eukaryotes and OsMADS29 is able to target certain regulatory components of this pathway for shifting the hormonal balance in favor of cytokinins.
PM464 (IPPC0295) Rice SKP-like gene, a component of E3 ligase, is involved in regulating pollen development and anther dehiscence Mrinalini Chawla1*, Meenu Kapoor2 and Sanjay Kapoor 1Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India 2University School of Biotechnology, Guru Gobind Singh Indraprastha University, New Delhi-110075, India *Presenting author: [email protected] Meiosis is an important event in the life of plants, as it marks the time of developmental transition from the sporophytic to the gametophytic phase. A comprehensive transcriptomics analysis using our in house data representing different stages/ tissues of vegetative and reproductive development in rice had revealed an important role of gene inactivation along with that of specific genes activation during male gametophyte development. We also found a marked increase in the activity of genes involved in ubiquitin-mediated protein degradation pathway. Central to this protein turnover machinery is the Skp1-Cullin-F-box (SCF) class of E3 ligases, which identify and prepare specific proteins for degradation by adding ubiquitination marks. Our investigations revealed 23 F-box and five OSK encoding genes expressing specifically in meiotic anthers in rice. Here, we show that silencing of one of these OSKs results in anther indehiscent phenotype and severe reduction in pollen viability. Silencing of this SKP gene does not affect the process of meiosis as well as mitotic divisions in the microspores. However, the microspores in OSK-KD lines remain smaller (by ~26%) in size in comparison to those in the wild type and fail to accumulate starch at the time of maturation, causing loss of viability. Furthermore, the endothecial layer of anther wall fails to develop the characteristic lignin striations, which have been shown to provide necessary mechanical pressure/tensile force required during anther dehiscence. These results highlight novel and essential roles of this gene in the 26S proteasome complex-mediated protein degradation during male gametophyte development.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 350 Session M : Reproductive Biology
PM465 (IPPC0298) Transcriptional properties of five seed-high NAC transcription factors Pinky Agarwal*, Iny Mathew and Akanksha Panwar National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Five NAC genes, which encode for transcription factors (TFs), express at high levels during rice seed development. This is reiterated by promoter-reporter rice transgenic plants. Rice genotypes with variable seed weights show variation in the expression patterns and levels of these genes during seed development. The promoter sequences of these genes, from different genotypes, show limited correlation with the expression. In terms of transcriptional properties, only one of the proteins is nuclear localized. However, many of these proteins dimerize amongst themselves and are subsequently nuclear localized. They are not only transcriptional activators, but strong repressors as well. Amongst the five NAC genes, one is alternatively spliced while two others are transcriptionally fused. Interestingly, the multiple forms of these genes have variable transcriptional properties. Summarily, the five seed-high NAC TFs and their forms have overlapping as well as distinct transcriptional properties, indicating their multiple roles in rice seed development.
PM466 (IPPC0358) PROTEIN L-ISOASPARTYL METHYLTRANSFERASE (PIMT) is implicated in seed desiccation tolerance and seed longevity in rice Nitin Kamble*, Bhanu Petla and Manoj Majee National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
PROTEIN L-ISOASPARTYL O-METHYLTRANSFERASE (PIMT) is a protein repairing enzyme and catalyses the conversion of spontaneously modified isoAsp to Asp in protein. The present study in rice demonstrates that PIMT activity sharply increases at maturation phase, retains in dry seed and then rapidly decline upon completion of germination. Likewise, deleterious isoaspartyl accumulation also increases during seed maturation and is highly abundant in dry seed but decreases upon imbibition. Western blot analyses clearly demonstrated distinct tissue and seed development stage specific accumulation of these PIMT isoforms, indicating their participation and specific contribution in seed longevity and germination in rice. For further analysis, we raised transgenic lines for each isoform and the data reveals the distinct roles of each OsPIMT isoform in restricting deleterious isoAsp accumulation to improve seed vigor and longevity. Collectively, these data imply that PIMT mediated protein repair mechanism initiates during the course of seed development in rice and each PIMT isoform plays a distinct yet coordinated role in maintaining seed vigor and longevity by restricting deleterious isoAsp accumulation in tissue and development stage specific manner.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 351 Session M : Reproductive Biology
PM467 (IPPC0369) Differentially expressed galactinol synthase gene (s) of chickpea (Cicer arietinum L.) improves seed vigor and longevity by limiting the hydroxyl radicals during aging Prafull Salvi* and Manoj Majee Plant Molecular Biology and biochemistry, National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected] Galactinol synthase (EC: 2.4.1.123) (GolS) catalyzes the rate limiting step in Raffinose Family Oligosaccharide (RFO) biosynthetic pathway. RFOs including galactinol are known as anti-stress agents and suggested to play important role in abiotic stress responses and seed desiccation tolerance. In this study, we have shown galactinol synthase activity as well as galactinol content is highly concentrated in pod and dry seed and gradually decreased as seed germination progresses. Galactinol synthase activity was also found to be enhanced in response to stresses. In chickpea, galactinol synthase enzyme is encoded by two divergent genes (CaGolS 1 and CaGolS 2) which produce 5 different types of transcripts. Transcription analysis revealed that the CaGolS1 and CaGolS2 are differentially regulated. Further, all isoforms were bacterially expressed, purified and biochemically characterized. All isoforms exhibit differential enzyme activity. Native molecular weight and subunit composition was determined for enzymatically active isoform through gel filtration chromatography. Confocal microscopy showed that a transiently transfected CaGolS-YFP fusion proteins primarily distributed to nucleus and membrane in onion epidermal cell. Further we found that seed-specific expression of CaGolS1 and CaGolS2 in Arabidopsis play a role in maintaining seed vigor and longevity, which was correlated well with increased galactinol and raffinose contents and reduced hydroxyl radicals in seed of transgenic lines. Collectively, our work clearly demonstrates that CaGolS(s) are differentially regulated in chickpea and play important role in enhancement of seed vigor and longevity.
PM468 (IPPC0485) Overexpression of OsbHLH142 causes complete male sterility in rice Rajeev Ranjan1*, Reema Khurana2 and Akhilesh Tyagi1 1National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India 2Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India *Presenting author: [email protected] Investigations on male sterility in crop plants are important for the hybrid seed production which provides improved yield as a result of heterosis. Rice is an important crop plant which is the source of food for almost half of the world population. Anther development in rice has been studied for many years which led to the identification of many molecular players. Yet, detailed mechanism associated with it remains elusive. Previously, we reported a bHLH transcription factor OsbHLH142 as an anther specific gene in rice. Now, we aimed to functionally characterize this during anther development in rice. Expression analysis of OsbHLH142 transcripts and polypeptide showed delayed accumulation of its protein compared to transcripts. Overexpression of OsbHLH142 in transgenic rice results in complete sterility and no seed set was observed. Crossing of transgenic plants with wild type pollen results in restoration of seed formation. Detailed observation of reproductive organs of transgenic plants revealed that anthers did not dehisce and most of the pollen grains were shrunken, abnormal shaped and non-viable. Electron microscopic and paraffin based tissue section analysis revealed detailed histological reason for these defects. RNA-Seq based transcriptome analysis of early and mature anthers suggests OsbHLH142 directly or indirectly regulates expression of different set of genes in the two stages of anthers. Moreover, through yeast two-hybrid and BiFC assays we have identified some OsbHLH142 interacting proteins which gives more insight on its molecular functions. Therefore, this study has developed new male sterile rice plants and explored novel functions of OsbHLH142 during rice anther development which will essentially help in the control of male fertility in crop plants.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 352 Session M : Reproductive Biology
PM469 (IPPC0508) Effects of auxins and other hormones on rice OsMADS29 expression Ridhi Khurana* and Sanjay Kapoor Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India *Presenting author: [email protected]
Rice MADS29 is a seed-specific MADS-box transcription factor that has been implicated in embryo and endosperm development. The OsMADS29 expression has been found to be tightly regulated at transcriptional, post-transcriptional as well as post-translational levels. We have previously demonstrated that OsMADS29 regulates cytokinin levels at the site of its action, not only in rice but also in heterologous tobacco BY-2 cells. In rice, the MADS29 transcripts begin to accumulate in high amounts soon after pollination, however, molecular basis of this induction is not known. Careful inspection of rice MADS29 upstream regulatory region (URR) revealed a number of auxin response elements (auxREs), besides a number of other conserved regulatory motifs. The auxREs are six base pair conserved motifs (TGTCTC) found in multiple tandem repeats in the promoters of early auxin response genes like GH3 and SAURs. And, the auxREs have been shown to be the targets of the DNA-binding domain of auxin response factors (ARFs). We have developed a tobacco BY-2 cell-line based assay system, where the transcriptional activity of OsMADS29URR::GFP under different hormonal regimes can be analysed. The results of these analyses will be discussed.
PM470 (IPPC0524) Downregulation of a seed-preferential mediator (MED) subunit affects reproductive development in rice Naveen Malik*, Pinky Agarwal and Akhilesh K. Tyagi National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
Mediator, a multi-subunit protein complex in eukaryotes, assists GTFs in RNA polymerase II mediated transcription by acting as a co-regulator. The whole mediator complex is structurally organized in head, middle, tail and CDK-cyclin C modules. Direct interaction of the tail module subunits with various transcription factors facilitates relay of signal to RNA pol II transcription machinery through head and middle modules. There is growing evidence for important roles played by various mediator subunits in Arabidopsis, but in rice the complex still remains structurally and functionally unexplored. In past, our group reported several mediator subunit genes having high expression during rice seed development stages and we chose one of these subunits for functional characterization. To unravel the function of this gene in rice, both overexpression and knockdown transgenic plants were raised. Overexpression plants were found to behave normally in most aspects of development and yield. On the other hand, knockdown plants showed various defects during reproductive development like non-viable pollen, reduced seed set and defective seed filling. Progress of the work on functional delineation of this MED subunit will be presented.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 353 Session M : Reproductive Biology
PM471 (IPPC0659) Various properties of a seed-preferential NAC transcription factor in rice Iny Mathew* and Pinky Agarwal National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected]
NACs are a large group of plant-specific transcription factors (TFs), playing important roles in plant life. All these proteins contain a highly conserved NAC domain identified initially in NAM, ATAF1/2 and CUC2 TFs, giving rise to the acronym ‘NAC’. In our microarray analysis, one member of this family has been shown to have seed-preferential expression, indicating its significance in seed development. Further, Q-PCR analysis confirms a high expression of this gene in rice seed developmental stages of five different genotypes with varying seed weight. The specific expression of the gene in the seeds can be credited to its 2 Kb promoter region as evident by the promoter-GUS analysis of transgenic plants. Alternative splicing has resulted in three different forms (A, B and C) of the transcript, which show difference in their C- terminal transcriptional regulatory region. Although, these proteins by themselves are localized in the cytoplasm, interaction with another NAC TF, recruit the A form to the nucleus as shown by BIFC experiments. Further, the different forms also show varying levels of transactivation ability. Over expression of this protein in planta points to the necessity of maintaining the temporal expression of this gene at an optimal level, as these plants show a slower growth rate with weaker and thinner aerial parts. The poster will focus primarily on the functional aspects of this gene in rice seed development.
PM472 (IPPC0666) Stigma and stylar movements vis-a-vis pollination dynamics in two medicinally important threatened plants Susheel Verma* and Ankush Khajuria Centre for Biodiversity Studies, Baba Ghulam Shah Badshah University, Rajouri-185232, J&K, India *Presenting author: [email protected]
Reproduction in flowering plants is a complex process involving several steps of communication between male and female counterparts. Many a times these interactions are mediated by agents both biotic and abiotic. Obviously the mechanisms underlying these interactions must be mediated by simple to complex signaling mechanisms operating either singly or interwoven. Two threatened and prized medicinal angiosperms namely Veleriana wallichii and Incarvellia emodi accomplish their mating in beautifully controlled ways. At the physical level, the manifestations are interesting and constitute the matter for this presentation. Flowers of Valeriana wallichii under pollinator limiting conditions fetch pollen themselves by moving their styles towards the pollen source, either within or in the neighboring flower. This way their stigmata get pollinated and reproduction is ensured. Stigma of Incarvillea emodi waits for requisite number of pollen to land on its surface even upto 9 days and then closes its lobes permanently. These mechanisms are responsible for successful sexual reproduction in these threatened taxa. Their control, however, remains a mystery.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 354 Session M : Reproductive Biology
PM473 (IPPC0682) Courtship in plants! What initiates it? Namrata Sharma* and Veena Sumbria Department of Botany, University of Jammu, Jammu-180006, J&K, India *Presenting author: [email protected]
Concealment of ovules inside the ovary has made pollen-pistil interaction an inseparable part of sexual reproduction in angiosperms. In most cases, this interaction initiates with the release of pollen from the anthers and their landing on the receptive part of pistil, i.e., stigma. This landing may be autonomous because of the close proximity of the two structures as happens in many autogamous taxa, or may involve a vector in cross-pollinated ones. Pistil in all these cases remains static structure while the pollen is mobile. During our work on the pollination mechanism in several flowering plants, we have encountered several cases, which defy these rules (e.g., in-situ pollen germination in Fumaria indica). Herein we present a recently encountered case in Tephrosia purpurea, wherein pistil exhibits beautiful movements to capture the pollen, which is being released by the anthers situated below the level of stigma. Although stylar movements have been reported previously also, the present study shows definitive phases of interaction between pollen and stigma facilitated by regular movements of both stigma and style. Prior to successful fertilization what stimulates these movements remains an open question. We speculate some sort of signalling mechanism operative in these movements. It is an endeavour for us to present these examples before the learned forum of plant physiologists to get a clue to the possible mechanisms involved in such interactions.
PM474 (IPPC0784) Phylogeny, expression and interaction of NF-Y TF family in rice Sweta Das*, Swarup Kumar Parida, Pinky Agarwal and Akhilesh Kumar Tyagi National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi-110067, India *Presenting author: [email protected] Nuclear factor-Ys (NF-Ys) are histone-like transcription factors which are present in all eukaryotes. These consist of three subunits, namely, NF-YA, NF-YB and NF-YC. These are encoded by single genes in yeast and animals but multiple number of genes in plants. NF-YB and NF-YC share sequence homology with H2B and H2A, respectively. They form dimers to which, NF-YA binds forming a trimer which in turn recognizes and binds to CCAAT box in the promoters. In plants, due to the presence of multiple subunits in each sub-class, there are manifold possibilities of these combinatorial complexes, which eventually lead to the functional diversity of NF-Ys. In rice, a total of 40 genes have been found to encode NF-Y TFs among which 11 code for NF-YA, 13 for NF-YB and 16 for NF-YC. Phylogenetic analysis shows homology of some of these genes with certain Arabidopsis NF-Ys and yet a few are unique to rice. Microarray data suggests that the expression of several of these TFs is constitutive and a few genes express in a seed specific/preferential manner. SNPs in 10 of these genes have been found to associate with seed size traits. These genes further exhibited differential expression patterns in seed developmental stages of rice genotypes with contrasting seed features. Study of transactivation properties of these TFs has revealed that few subunits belonging to each of the sub-class positively regulate transcription. Yeast two-hybrid interactions amongst OsNF-Ys suggest interaction of certain subunits with each other. An overview of the OsNF-Y family will be presented here.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 355 Session M : Reproductive Biology
PM475 (IPPC0788) Understanding sex expression and modification in Coccinia grandis - A dioecious member of Cucurbitaceae Ravi Devani1*, Amita Ghadge1, Kanika Karmakar2, Jayeeta Banerjee1, Sangram Sinha2 and Anjan Banerjee1 1Indian Institute of Science Education and Research, Pune-411008, Maharashtra, India 2Department of Botany, Tripura University, Suryamaninagar-799022, Tripura, India *Presenting author: [email protected]
Majority of the angiosperm species are co-sexual (Hermaphrodite or Monoecious). Only about 6% of angiosperm species are dioecious having separate male and female individuals. From the phylogenetic distribution of dioecious species, it is clear that dioecy has evolved many times in plant kingdom independently with multiple molecular mechanisms governing sex expression. Also, a few dioecious species have further evolved sex chromosomes like most of the mammalian species. Dioecious systems also exhibit a certain degree of lability in sex expression. Chemical treatments like AgNO3 and Ag2S2O3, treatment with some phytohormones, and hypomethylating agent 5-azacytidine are known to alter sex expression in dioecious plants. Coccinia grandis is one such dioecious species having heteromorphic sex chromosomes. C. grandis belongs to Cucurbitaceae, a family well known for its diverse sexual systems. Availability of four sequenced genomes in Cucurbitaceae gives C. grandis, an advantage as a model system to study dioecy. Sex modification has been studied in hypogynous Silene latifolia (Caryophyllaceae) but there are no such reports in epigynous Coccinia grandis. We have taken a candidate gene approach to understand the role of flowering related genes in the development of unisexual flowers of C. grandis. Simultaneously, a comparative transcriptomic approach has been initiated to identify novel players involved in sex expression and modification. Investigations on sexual phenotypes of C. grandis including a rare gynomonoecious (GyM) form and AgNO3 mediated sex modification will add a new dimension to the understanding of sex expression in dioecious flowering plants.
PM476 (IPPC0852) Expression and promoter analysis of seed storage protein encoding genes in rice Akanksha Panwar*, Iny Elizebeth Mathew and Pinky Agarwal Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi-110021, India *Presenting author: [email protected]
In monocots such as rice, seed storage proteins (SSPs) are synthesized in the endosperm during seed development, which are essential source of proteins in human diet. On the basis of solubility, seed storage proteins are classified into four categories: albumins (soluble in water), globulins (soluble in saline), prolamins (soluble in aqueous alcohol) and glutelins (soluble in alkali). Glutelin is a major SSP in rice, present in 60-80% by weight of the total protein, whereas, prolamin contributes to 20-30% of total proteins. Glutelins in rice have been reported to be homologs of 11S globulins of legumes, while prolamins have been shown to be originated from an alpha-globulin storage protein. Endosperm-specific expression of SSP genes is regulated by cis-elements present in their promoters, such as GCN4, a prolamin box (P-box), AACA, ACGT. These motifs act as binding sites for TFs to regulate their expression, of which many are known to be conserved. We have attempted to identify SSP encoding genes of rice and classify them phylogenetically, along with their promoter and expression analysis.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 356 Session M : Reproductive Biology
PM477 (IPPC0886) Associated bacteria assist in reproduction of seaweed, Gracilariadura Ravindra Pal Singh* Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, 6-10-1, Fukuoka, 812-8581, Japan *Presenting author: [email protected]
Seaweed associated bacteria play a crucial role in morphogenesis and the growth of seaweeds in direct and/or indirect manners. Bacteria of phylum proteobacteria and firmicutes are generally most abundant on the seaweed surface. Associated bacteria produce hormones, quorum sensing signals, bioactive compounds and other effective molecules that are responsible for normal morphological structure and development in seaweeds. Zoospores of Ulva species respond to N- acyl-homoserine lactones (AHL) produced by gram- negative bacteria and settle near the centre of AHL production. In this regards, we isolated several gram-negative bacteria from different seaweeds and out of them seven isolates were found to produce different types of N-acyl homoserine lactones (AHLs). Interestingly, Shewanella algae produced five types of AHL: C4-HSL, HC4-HSL, C6-HSL, 3-oxo-C6-HSL and 3-oxo-C12-HSL. Subsequently, the AHLs producing bacterial isolates were screened for carpospore liberation from G. dura and these isolates were found to positively induce carpospore liberation over the control. Also, we observed that carpospore liberation increased significantly in C4- and C6-HSL treated cystocarps. Sodium dodecyl sulfate and native polyacrylamide gel electrophoresis of the total protein of the C4- and C6- HSL-treated cystocarps showed two specific peptide bands of different molecular weights (50 and 60 kDa) as compared to the control, confirming their indirect effect on carpospore liberation.
PM478 Effect of high temperature stress on physiological attributes, antioxidative enzymes and expression of heat shock proteins in wheat (Triticum aestivum L.) N.K. Gupta*, Aarif Khan and R. S. Sain Seed Technology Research, Rajasthan Agricultural Research Institute, Durgapura- Jaipur-302018, Rajasthan, India *Presenting author: [email protected]
Experiment was conducted to study the effect of heat stress on various parameters in five diverse wheat genotypes at seedling stage. Seeds were sown in plastic pots in laboratory at 25±2°C. For heat treatment, 20 days old seedlings were exposed to 36°C/42°C for 6 hours. Results showed that the heat stress significantly reduced the dry weight and length of seedlings in all the genotypes. Chlorophyll content and membrane stability index decreased but proline content increased in all the genotypes under stress conditions. Among genotypes, Raj 4083 exhibited better heat tolerance, particularly at 42°C. Activities of SOD, CAT, APX and POX increased significantly in all the genotypes but per cent enhancement was maximum in Raj 4083 and minimum in PBW 502. In SDS-PAGE protein profiling, 15 polypeptides were recovered in control plants with slight variations in temperature treated plants of most genotypes. Proteins with molecular weights of 85 kDa and 15 kDa were constitutively expressed in all the genotypes but proteins of 45 kDa were expressed only in tolerant genotypes. Expression analysis of heat shock proteins, particularly hsp 90 further indicated the existence of tolerance mechanism in Raj 4083. Thus, using a multi parametric approach, Raj 4083 was found most tolerant and PBW 502 most susceptible to heat stress. The study also provides evidence that antioxidant enzymes like SOD, CAT, GPX, APX and heat shock proteins like HSP 90 have association with heat tolerance in wheat and thus can be used as efficient markers for heat tolerance at seedling stage.
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 357 ted bacteria assist in Author Index
Abass Nazia ...... 224 Anand Saurabh ...... 319 Baig K.S...... 132 AbdElgawad Hamada ...... 110 Ananya Chakrabarty ...... 140 Bains Gurdeep ...... 118 Abdin M.Z...... 39 Anders Nadine ...... 201 Bains Navtej Singh ...... 149 Abdul R...... 151 Anderssen Bob ...... 81 Bajaj Deepak ...... 104 Abdullah Hesham ...... 37 Andreuzza Bindu ...... 79 Bakshi Achala ...... 180 Ablordeppey Kenny ...... 37 Andreuzza S...... 102 Bal Santanu ...... 345 Achary Mohan ...... 239 Andrew Lynn ...... 344 Bala Kiran ...... 220 Achary Mohan Murali ...... 77 Angadi Sangamesh ...... 149 Bala Suman ...... 260 Achary V. Mohan ...... 225,240,315 Ansari Mohammad Wahid ...... 118 Balaji M...... 17 Acharya Vishal ...... 57,340 Ansari Shamim ...... 316 Balyan Sonia Chhillar ...... 12 Adhikari Kailash ...... 65 Antony Edna ...... 275 Bamel Kiran ...... 285 Adhikari Kedar ...... 121 Anuj Abhishek Kumar ...... 314 Banday Zeeshan Zahoor ...... 208 Aftab Tariq ...... 294 Anuradha M...... 124 Bandyopadhyay Anindya ...... 70 Agarwal Aakrati ...... 77,129,239 Anwar Khalid ...... 122,226 Banerjee Anjan ...... 189,356 Agarwal Aditya Vikram ...... 318 Archak Sunil ...... 156 Banerjee Jayeeta ...... 356 Agarwal Gaurav ...... 216 Argal Surendra ...... 276 Banerjee Kaushik ...... 344 Agarwal Manu ...... 17,51,176 Ariyadasa Thilini ...... 122 Banerjee Nirupama ...... 253 Agarwal Meenakshi ...... 242 Aro Eva Mari ...... 109 Bangar Pooja ...... 154 Agarwal P...... 43 Arora Ajay ...36,127,132,143,144,286,287 Bannerjee K...... 102 Agarwal Pallavi ...... 182 Arora Priyanka ...... 181 Bansal K.C...... 7,156,326 Agarwal Parul ...... 317 Arora Sandeep ...... 166 Banu Sofia ...... 251 Agarwal Pinky ...... 351,353,354,355,356 Aruna C...... 130 Bapatla Ramesh B...... 67,229 Agarwal Preeti ...... 147 Arya Deepshikha ...... 35 Barchi Boubker ...... 8 Agarwal R.M...... 276 Arya Gulab ...... 190 Bareth S.S...... 273 Agarwal Surekha Katiyar ...... 17,176 Arya Preeti ...... 340 Barik Jayanta ...... 293 Aggarwal P.K...... 156 Arya R.R...... 160 Barman Paramananda ...... 218 Agurla Srinivas ...... 187 Asard Han ...... 110 Barnabas A.D...... 336 Ahamed M. Lal ...... 174 Ashish Subba ...... 122 Barua Praga ...... 207 Ahanger Mohammad A...... 276 Ashraf Nasheeman ...... 223,224 Baruah K.K...... 62,283,297,305 Ahlawat Yogesh ...... 214 Ashwini N...... 198 Baruah Sunitee Gohain ...... 305 Ahmad Altaf ...... 309 Asif M.H...... 299 Basak Sandip Kumar ...... 141 Ahmad Syed Sabhi...... 83 Asif Mehar ...... 205,317 Basu P.S...... 143 Ahmad Tauqueer ...... 335,338 Asrey Ram ...... 335 Basu Partha ...... 272 Ahmadi N...... 99 Aswani Vetcha ...... 67 Basu Saumik ...... 47 Ahmadizadeh Mostafa ...... 103 Atkins Peggy Ozark ...... 199 Basu Urmila ...... 312 Ahsan Hafiza ...... 270 Atri Anupama ...... 332 Basumatary Nabami ...... 176 Akbari Parisa ...... 37 Auguin Daniel ...... 9 Batra V...... 161 Akula Shalini ...... 177 Augustine Rehna ...... 41 Bechtold Ulrike ...... 32,123,136 Alharbi Najeeb ...... 121 Avasthi Uday Kiran ...... 306 Beckmann Hauke ...... 201 Alivelu K...... 142 Awakale Pramod ...... 190 Bedi Seema ...... 332 Aloor Bindu Prasuna ...... 306 Azeez Abdul ...... 299 Beemster Gerrit T.S...... 110 Alvarez-Fernandez Ruben ...... 69 Baba Shoib ...... 224 Begam Momtaj ...... 141 Amardeep ...... 123 Babita...... 210 Begna Sultan ...... 149 Amaregouda A...... 131,264 Babitha K.C...... 45 Behera Tusarkanti ...... 315 Amarshettiwar S.B...... 152 Babu O. Sudhakar ...... 133 Bender Kyle ...... 114 Amasino Richard M...... 44 Baburam V...... 239 Bertheau Lucie ...... 9 Anand Abhishek ...... 247 Bagri Deepak Singh ...... 262,281 Best Norman ...... 75 Anand Anjali ...... 145 Bahuguna Rajeev K...... 11 Bethoju K...... 102
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 359 Author Index
Bhaduri Debarati ...... 159 Bohra Burhanuddin...... 280 Chaudhary Chanderkant ...... 219 Bhagat Kiran ...... 345 Boni Rainer ...... 243 Chaudhary Saurabh ...... 28 Bhagat Prakash Kumar ...... 185 Boominathan P...... 154 Chaudhury Ashok ...... 154,220 Bhalla Prem L...... 73,80 Bora Ranjan Kumar ...... 176 Chauhan D.A...... 168 Bhambhani Sweta ...... 317 Borah Leena ...... 297 Chauhan Harsh ...... 243 Bhandari Pinki ...... 242 Borah Pratikshya ...... 218 Chauhan P.S...... 182 Bhandari Purnima ...... 265 Bordoloi Nirmali ...... 283 Chauhan Rajinder Singh ...... 337 Bhandawat Abhishek ...... 221,325 Borphukan Bhabesh ...... 225,239,240,315 Chauhan Rohit ...... 57 Bharadwaj Chellapilla ...... 323 Brahma Vijya ...... 220,340 Chaurasia Akhilesh Kumar ...... 331 Bharali Ashmita ...... 62,305 Bramley H...... 95,121 Chawla Mrinalini ...... 114,350 Bhardwaj Archana ...... 210 Brignolas Franck ...... 9 Chawla Vandna ...... 57 Bhardwaj Chhavi ...... 220 Burman Naini ...... 192,193 Cheema H.S...... 273 Bhardwaj Mansi ...... 49 Burman Uday ...... 157,348 Chefdor Françoise ...... 9 Bhardwaj Vasudha ...... 314 Burondkar Murad ...... 85,284,306,324 Cheong Yong-Hwa...... 15 Bhargav R...... 120 Calapit-Palao Cecilia Diana O...... 103 Chetti M.B...... 82,284 Bharti Amrit ...... 134 Carlos Guzman...... 318 Chettry Upasna ...... 217 Bharti Pooja ...... 328 Carolina Paola Sansaloni...... 318 Chevala Narayana ...... 211 Bhaskar Avantika ...... 186 Carolina Saint-pierre ...... 318 Chhajer Sidhika ...... 283 Bhaskar Yogendra ...... 202 Carpin Sabine ...... 9 Chhapekar Sushil...... 220,328,344 Bhasker Pattipaka ...... 177 Chaitanya Bharatula ...... 231 Chhikara Sudesh ...... 37 Bhat Hilal A...... 113, 347 Chakrabarti B...... 277 Chhuneja Parveen ...... 112 Bhat K.V...... 154,326 Chakrabarty Broto...... 233 Chikani B.M...... 302 Bhat Prabhakar ...... 130 Chakraborty Anirban ...... 155 Chinreddy Subramanyam ...... 129 Bhat Ravi...... 100 Chakraborty Koushik...... 72,159 Chitikineni Annapurna ...... 217,322 Bhatia Arti ...... 278 Chakraborty Navjyoti ...... 55,191 Chitwood Daniel ...... 110 Bhatia Chitra ...... 206 Chakraborty Niranjan..131,133,152,196, 207 Chopra A.K...... 169 Bhatia Sabhyata ...... 200,203,316 Chakraborty Subhra ...... 133,152,207 Chopra Preeti ...... 272 Bhatnagar Akanksha ...... 192,193 Chakraborty Supriya ...... 49,245 Chopra Renu Khanna ...... 1,134 Bhatt R.K...... 283,348 Challabathula Dinakar ...... 139 Choubey Ami ...... 289 Bhatt R.M...... 343 Chamanthula Gayathri ...... 339 Choudhary Chetan Kumar ...... 140 Bhattacharjee Annapurna ...... 211 Chand Gurdev ...... 177 Choudhary Harshawardhan ...... 315 Bhattacharjee Payel ...... 259 Chand Mehar ...... 135 Choudhary R.L...... 120,276 Bhattacharjee Saikat ...... 256,257,258 Chandra A...... 262,271 Choudhary Ritika ...... 140 Bhattacharrya Abhishek ...... 33 Chandra Amaresh ...... 155,222 Choudhur Devapriya ...... 55 Bhattacharya M.K...... 163 Chandra Deepak ...... 318 Choudhury P.P...... 345 Bhattacharyya Dhriti ...... 245 Chandran Divya ...... 256,257 Choudhury Swarup Roy ...... 18 Bhave Sanjay ...... 85,284,324 Chandrapal...... 296 Chowdappa P...... 100 Bhisth Kavitha ...... 112 Chandrasekhar K...... 129 Chowdhury A. Roy ...... 331 Bhowmick P.K...... 74,320 Changwal Chunoti ...... 286,287 Chowdhury Rajojit ...... 141 Bhushan Deepti ...... 131 Chatrath Ravish ...... 140 Christopher Penfold ...... 69 Bhyri Priyanka ...... 79 Chattha Yadvendra Singh ...... 186 Chrungoo N.K...... 21,217,322,323 Birbal...... 120 Chattopadhyay Debasis ...... 207,252,319 Clerget Benoit ...... 264 Bishi S.K...... 42,159,255,302 Chattopadhyay Sudip ...... 13,320 Clevenger Josh ...... 199 Bisht Naveen ...... 41,190 Chaturvedi Arjit ...... 227 Courdavault Vincent ...... 9 Biswal Basanti ...... 71 Chaturvedi Ashish K...... 98,159,169 Cressman Kurtis ...... 199 Biswas Ashish ...... 268 Chaturvedi S.K...... 272,322 Dadlani Malavika ...... 230 Biswas Asok Kumar ...... 225 Chaudhari Vidhya ...... 269 Dafyd Jenkins ...... 69 Bobba Sunil ...... 305 Chaudhary Bhupendra ...... 191 Dagla M.C...... 302
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 360 Author Index
Dahuja Anil ...... 338 Dhandapani R...... 288 Gajjeraman Prabu ...... 259,341 Dalal Girish ...... 280 Dhankher Om Parkash ...... 37 Gali S.K...... 275 Dalal Monika ...... 190,282,324 Dhanoji M.M...... 131,277 Ganapathi Thumballi ...... 266,267 Dalvi D.G...... 132 Dhansu Pooja ...... 135 Gandotra Vandit ...... 209 Dalvi Vijay ...... 324 Dhanyalakshmi K.H...... 326 Ganesh S. Sree ...... 168 Damodarreddy D...... 124 Dhar Swatismita ...... 241,255 Ganie Showkat Ahmad ...... 128,212 Damor Parthvee R...... 168 Dharanipragada Prashanthi ...... 233 Garai Sampurna ...... 68 Dar M.A...... 270 Dharmaraj P.S...... 217 Garg A...... 172 Das Amit ...... 79 Dhawan Gaurav ...... 320 Garg Neera ... 125,134,173,179,265,266,271 Das Anup ...... 349 Dhepe A.P...... 311 Garg Rashmi ...... 301 Das Ayan...... 246 Dhingra Anuradha ...... 155 Garg Rohini ...... 111,211,212 Das Bappa ...... 92,343 Dhuppar Pooja ...... 189,191 Garg Vanika ...... 216,217 Das M...... 171 Dilkes Brian ...... 75 Garggi G...... 226 Das Madhumanti ...... 184 Dinesh Rachana...... 292 Gaur Asha ...... 262,271 Das Madhurima ...... 233 Dingkuhn M...... 99 Gaur Pooran M...... 215,322,323 Das Priyanka ...... 11,116,165 Dixit Sandeep ...... 246 Gaur Rashmi ...... 220,328,344 Das Rohit ...... 259 Djeghdir Inès ...... 9 Gautam Anuradha ...... 74,297 Das Roma ...... 323 Dkhar Jeremy ...... 202 Gautam Janesh Kumar ...... 320 Das S...... 43 Doddamani Dadakhalandar ...... 216,322 Gautam Manisha ...... 171 Das Sampa ...... 246 Doddamani M.B...... 275 Gayatri Gunja ...... 187,188 Das Sandip ...... 319 Doddaraju Pushpa ...... 234 Geeta R...... 218 Das Satyajit ...... 160 Dohtdong Lashaihun ...... 323 Geetha Gouribidanur A...... 106 Das Shouvik ...... 104 Donald James ...... 240,315 Gera Rajesh ...... 261 Das Sweta ...... 355 Dutt Som ...... 236 Ghadge Amita ...... 356 Dash S.K...... 343 Dutta Ayan ...... 33 Ghangal Rajesh ...... 28 Dasmunshi Anilabh...... 315 Dutta Tanima ...... 114 Ghanta Anuradha ...... 217 Datta Swapan K...... 322 Dutta Tushar Kanti ...... 251 Gharpure S...... 102 Davda J...... 102 Dwivedi Priyanka ...... 320 Ghate Tejashree ...... 189 David Wild ...... 69 Dwivedi S.K...... 116 Ghazi Irfan A...... 235 Dayanandan P...... 336 Dwivedi Varun ...... 235 Ghosh Ajit ...... 64 Debnath Suprokash ...... 174 Dwivedi Vikas ...... 319 Ghosh Ananta Kumar ...... 150 Deokate P.P...... 276 Ekamber Kariali ...... 290 Ghosh Daipayan ...... 175,177 Deosarkar D.B...... 132 Elanchezhian R...... 268 Ghosh Indira ...... 91,233 Depierreux Christiane ...... 9 Ellur R.K...... 74 Ghosh Jyotirmoy ...... 331 Desai B.K...... 264 Ewers Brent E...... 44 Ghosh Shraboni ...... 137 Deshmukh Nishant Anandrao ...... 349 Fareed Mohd. Khan ...... 67 Ghosh Sourav ...... 176 Deshpande S.P...... 101 Farhi Moran ...... 110 Ghosh Srayan ...... 243,248 Dev Brihama ...... 315 Fartyal Dhirendra ...... 77,129,239,315 Ghosh Zhumur ...... 259 Devadasu Elsin Raju ...... 309 Faseela P...... 303 Gill Kulvinder Singh ...... 318 Devadiga Pooja ...... 241 Ferguson John ...... 32 Giri Jitender ...... 250,263,313 Devani Ravi ...... 356 Finnegan Jean...... 81 Giri Mrunmay ...... 320 Devanna B.N...... 22 Forster Genevieve M...... 235 Glévarec Gaëlle ...... 9 Devi Ayam Gangarani ...... 21 Foureau Emilien ...... 9 Gnanaraj Muniraj ...... 291 Devi Sarita ...... 177 Francis Aleena ...... 203,314 Gnanasekaran Prabu ...... 245 Devmore Jalindar ...... 324 Fulzele Devanand P...... 139 Godara O.P...... 144 Dey Narottam ...... 321 Gadag R.N...... 169 Goel Parul ...... 57,273 Dey Sayani ...... 259 Gadi V.K...... 172 Goel Ridhi ...... 299 Dhakar R...... 127,144 Gaikwad Kishor ...... 216 Goel Shailendra ...... 51,176
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 361 Author Index
Gohain Bornali ...... 258 Harishukumar P...... 267 Jayaswall Kuldip ...... 325 Gohara Elizabeth ...... 90 Harit R.C...... 277 Jerez Ivone Torres ...... 199 Gokhale Nitin ...... 284 Haritha B...... 74 Jethva Minesh ...... 94,335 Golakiya B.A...... 42 Hashimoto Kenji ...... 34 Jeyakumar P...... 282,292,295 Gopalakrishnan S...... 272 Hassan Mohd ...... 240 Jeyapriya S...... 308 Goplakrishnamurthy T...... 124 Hebbar K.B...... 100 Jez Joseph ...... 58 Goswami Nisha ...... 72,159 Helliwell Christopher ...... 81 Jha Aditya Nath ...... 268 Govindasamy V...... 120 Hemamalini P...... 343 Jha Bhavanath ...... 232 Govindjee...... 86 Hemantaranjan A...... 115,286 Jha Gopaljee ...... 243,245,248 Greene Thomas W...... 79 Héricourt François ...... 9 Jha Gulshan ...... 62 Greenham Kathleen ...... 44 Hermand Victor ...... 44 Jha S.K...... 275,335 Grover Anil ...... 25,155 Hickey John M...... 323 Jha Sarita ...... 215 Grover Khulbhushan ...... 149 Hirt Heribert ...... 5 Jha Saroj K...... 113 Grover Minakshi ...... 177 Hooley Richard ...... 55 Jha Shweta ...... 250 Guerinot Mary Lou ...... 87 Hossain Firoz ...... 53,274 Jhansi K...... 311 Gujjarlapudi Mariyamma ...... 252 Hummel Grégoire ...... 313 Jian Jianbo ...... 322 Gulabani Hitika ...... 256 Hussain Aubid ...... 224 JoãoAraújo Jr, ...... 304 Gulati Ranju ...... 179 Ichihashi Yasunori ...... 110 Jodder Jayanti ...... 259 Gunjan...... 331 Igor Chernukhin ...... 69 Johal Gurmukh S...... 75 Gupta Aarti ...... 246 Imam Muhammad Ali ...... 342 Jondhale Ashish ...... 324 Gupta Aditi ...... 280,296 Ingole Kishor ...... 258 Jorge Franco...... 318 Gupta Anil ...... 149 Iqbal M...... 309 José Crossa...... 318 Gupta Asmita ...... 301 Islam Md Rofiqul ...... 251 Joshi Alka ...... 236 Gupta Brijesh K...... 27,121 Ismail Abdelbagi M...... 156 Joshi Babita ...... 263 Gupta Deepti ...... 196 Itabashi Etsuko ...... 73 Joshi Chandrashekhar P...... 214 Gupta H.S...... 53,274 Iyer Ganesh ...... 241 Joshi K.A...... 311 Gupta Maitrayee Das ...... 33 Jack Matthews ...... 69 Joshi Raj Kumar ...... 238,312 Gupta Megha ...... 257 Jadon K.S...... 302 Joshi Rohit ...... 27,281 Gupta N.K...... 273,357 Jadon Kuldeep Singh ...... 42 Juan Burgueno Ferrera...... 318 Gupta Nakul ...... 230 Jagadis Kapuganti ...... 194 Juergens Gerd ...... 201 Gupta Parul ...... 317,318 Jagannath Arun ...... 51,176 Julia C...... 99 Gupta Prakash Chandra ...... 279 Jain A...... 52 Jung Chol-Hee ...... 80 Gupta Priya ...... 222,249 Jain Ajay ...... 272 Kaiser Werner ...... 194 Gupta Priyanka ...... 119,188 Jain Ankit ...... 28,215 Kakani R.K...... 170 Gupta Raj Kumar ...... 346 Jain Mukesh ...... 24,211,212 Kakralya B.L...... 144 Gupta Rajendra ...... 285 Jain Neelu ...... 112,178,262 Kalariya K.A...... 72 Gupta Ramneek ...... 51 Jain Nitin ...... 162,193 Kale Manisha ...... 231 Gupta Shrish C...... 285 Jain Priyanka ...... 330 Kale Sandip ...... 215,217 Gupta V.K...... 126,335,338 Jain Radha ...... 155,222,262,271 Kalia Rajwant K...... 283 Gupte Yash ...... 241 Jainender ...... 135 Kalita Lotika ...... 349 Gurrapu Ramya ...... 233 Jaldhani V...... 321,307 Kalyani Tripathi ...... 268 Habib Farhat ...... 189 Jaleel Hassan ...... 291 Kambalimath S.K...... 234 Hairat Suboot ...... 290 Jalmi Siddhi ...... 244 Kamble Nitin ...... 351 Haldankar Parag ...... 306 James Beynon ...... 69 Kannababu N...... 130 Haloi Meghna ...... 349 James Donald ...... 129,239 Kannan K...... 137 Harikrishna ...... 112 Jangir Monikam ...... 342 Kant Chandra ...... 200,203 Hariprasanna K...... 130 Jannink Jean-Luc ...... 323 Kantiwal Usha ...... 220 Harish M...... 131 Janssens Ivan A...... 110 Kanwar Poonam ...... 15,243,248
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 362 Author Index
Kanyuka Kostya ...... 55 Khan Nafees A...... 279 Kulshreshtha Neeraj ...... 29,161 Kapare Vishwadeep ...... 139 Khan Sadaf ...... 299 Kumar Abhishek ...... 262 Kapoor Meenu ...... 35,350 Khan Zaiba Hasan ...... 209 Kumar Amar ...... 51,176 Kapoor Sanjay ...... 43, 78,114,350,353 Khandal Hitaishi ...... 207 Kumar Ambarish ...... 340 Kapuganti Jagadis ...... 195 Khandelwal Abha ...... 90 Kumar Amish ...... 336 Kar Rup Kumar ...... 160,213,214 Khangura Rajdeep S...... 75 Kumar Amit ...... 189 Karaba Nataraja N...... 45 Khare Ria ...... 205 Kumar Anil ...... 240 Karan Ratna ...... 183 Kharshiing Gayle ...... 322 Kumar Arun ...... 112 Kariali Ekamber ...... 293 Khatediya Nilesh ...... 42 Kumar Ashish ...... 28 Karmakar Kanika ...... 356 Khetarpal Sangeeta ...... 145,274 Kumar Ashwani ...... 135,316 Karn Rajni ...... 315 Khobra Rinki ...... 346 Kumar Atul ...... 118 Karuppanapandian Thirupathi ...... 291 Kholova Jana ...... 313 Kumar Bhumesh ...... 345 Karuputhula N.B...... 233 Khurana J.P...... 6,138,162,186,192, Kumar Dameshwar ...... 268 Karwa Sourabh ...... 159 193,194,218,219,333 Kumar Devendra ...... 126 kashyap Jyoti ...... 342 Khurana Paramjit...... 19,138,147,219, Kumar Dinesh ...... 145 Kashyap Lakita ...... 265 223,290 Kumar G. Vijaya ...... 151 Katiyar Deepmala ...... 286 Khurana Priyanka ...... 220 Kumar Gulshan ...... 210 Kaul Tanushri ...... 77,264 Khurana Ridhi ...... 353 Kumar H.G. Jalendra ...... 30,234,327 Kaul Veenu ...... 107 Kientz Marika ...... 201 Kumar Jitender ...... 118,260 Kaur Amarjot ...... 333 Kiep Victoria ...... 250 Kumar K. Sumanth ...... 180,327,329 Kaur Charanpreet ...... 64,112 Kim Boem-Gi ...... 15 Kumar Kamal ...... 221,234,258 Kaur Gundeep ...... 114 Kiran B.O...... 264 Kumar Krishan ...... 177 Kaur Harsimran ...... 114,332 Kiran K.R...... 195 Kumar Kundan ...... 162 Kaur Kanwaljeet ...... 191 Kiranmai K...... 133 Kumar Lokesh ...... 302 Kaur Navdeep ...... 167 Kirti P.B...... 180 Kumar M.B. Arun ...... 230 Kaur Parmeet ...... 123 Kitahata Nobutaka ...... 34 Kumar M.V. Mohan ...... 30,327 Kaur Preeti ...... 164 Kohli Ajay ...... 23 Kumar Mahesh ...... 120 Kaur Pritam ...... 51 Kondamudi Rajesh ...... 307 Kumar Mukesh ...... 12,253 Kaur Ranjeet ...... 150 Kondhare Kirtikumar ...... 189 Kumar Mukul ...... 279 Kaur Tarandeep ...... 113,347 Koppad Appanna V...... 334 Kumar Muthappa Senthil...... 246,247 Kaushal Jyoti ...... 297 Korasick David ...... 58 Kumar Narendra ...... 127,128,302 Kav Nataraj ...... 312 Kotvi Payal ...... 298 Kumar Neeraj ...... 177,178 Kavya Y...... 102 Krattinger Simon ...... 243 Kumar Pramod ...... 148,302,316 Kaya Hidetaka ...... 34 Krishna Bal ...... 331 Kumar Praveen ...... 157 Kelkar Kaustubh ...... 306 Krishna G...... 92,343 Kumar Rajeev ...... 235,245,335,338 Keller Beat ...... 243 Krishna G.K...... 171 Kumar Rajnish ...... 344 Kemisetti Siva Raju ...... 267 Krishnamurthy S...... 314 Kumar Rakesh ...... 92 Kevat Nisha ...... 167 Krishnamurthy S.L. 20,29,158,161,165,173 Kumar Ravi ...... 315 Khajuria Ankush ...... 354 Krishnan P...... 344 Kumar Reddy Kishore...... 245 Khan Aamir ...... 217 Krishnan S. Gopala ...... 74,230,320 Kumar Roshan ...... 190 Khan Aamir W...... 216 Krishnan Victor Banerjee ...... 344 Kumar S. Rajesh ...... 196 Khan Aarif ...... 126,357 Krishnapriya V...... 269,274 Kumar Sanjay ...... 2 Khan Akhtar Hussain ...... 156 Kryukov Lavr ...... 60 Kumar Santosh ...... 116 Khan F.A...... 270 Kuchitsu Kazuyuki ...... 34,187 Kumar Satish ...... 177 Khan Fareed ...... 230 Kudapa Himabindu ...... 216 Kumar Shiwesh ...... 101 Khan M. Iqbal R...... 279 Kudla Jörg ...... 15 Kumar Smita ...... 205 Khan M. Masroor ...... 294 Kujur Alice ...... 104 Kumar Sudhir ...... 142,147 Khan M. Masroor A...... 291,298 Kulkarni Mahesh ...... 85,306 Kumar Sumit ...... 231,303
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 363 Author Index
Kumar Sundeep ...... 165 Lekshmy S...... 275,324 Manga V.K...... 348 Kumar V...... 277 Liew Lim Chee ...... 80 Mangrauthia S.K...... 307 Kumar V.V. Santosh ...... 16,146,157, Lohot Vaibhav D...... 331 Manivannan A...... 348 164,171,172,189,191,282,324 Lokesh U...... 133 Manjunath Manju ...... 76 Kumar Vandna A...... 240 Lokya Vadthya ...... 252 Mann Anita ...... 269 Kumar Verandra ...... 210 Long Stephanie ...... 26 Manna Mrinalini ...... 77,239,315 Kumar Vijay ...... 278 Lorenz Aaron ...... 323 Manna Suman ...... 347 Kumar Vinay ...... 217 Lou Ping ...... 44 Mannully C.T...... 186,204 Kumar Vinod ...... 20,300 Louis Joe...... 47 Manoharan Kumariah ...... 291 Kumar Krishan ...... 178 Luan Sheng ...... 4,15 Maramreddy M.K...... 233 Kumara K. Sumanth ...... 197 Luiz Fernando Almeida...... 304 Maria Vera Da Costa ...... 310 Kumara P. Mohana...... 114 Lymm Andrew ...... 340 Marik Ananya ...... 184 Kumari A.P. Padma ...... 63 Lynn Andrew ...... 202,220 Masand Shikha ...... 117 Kumari Anita ...... 135 Macwan Sunil J...... 288 Mathew Iny ...... 351,354 Kumari Archana ...... 330 Madireddi S.K...... 309 Mathew Iny Elizebeth ...... 356 Kumari Asha ...... 181 Mahadik Sandip ...... 284 Mathew Reynolds...... 318 Kumari B. Ramana ...... 321 Mahajan Ameya ...... 189 Mathur P.N...... 156 Kumari Chanchal ...... 251 Mahajan Gulshan ...... 123 Mathur Saloni ...... 12,215 Kumari Jyoti ...... 165 Mahatma Lalit ...... 168 Mathur Supriya ...... 203 Kumari Poonam ...... 260 Mahatma M.K...... 42,255,302 Maurya Sadhana ...... 169 Kumari Punam ...... 253 Mahato K.K...... 195 Max K.F...... 102 Kumria Ratna ...... 79 Mahesh S...... 102 Mayee Pratiksha ...... 332 Kundu Pallob ...... 246,259 Mahesh Shashank ...... 316 Mayer Ulrike ...... 201 Kundu Siddhartha ...... 339 Maheswari M...... 96,135,151 McClung C. Robertson ...... 44 Kuriakose Saritha V...... 90 Maheswari Sellakumar ...... 291 Meena Hari Singh ...... 169 Kushwaha Hemant R...... 94,335 Mahindra Anuradha ...... 90 Meena M.K...... 82 Kushwaha N.K...... 49,245 Mahmood Saquib ...... 253 Meena Mukesh Kumar ...... 277 Kushwaha S.R...... 172,282,296 Mahto Binod Kumar ...... 241,255 Meena R.C...... 178 Lafarge Tanguy ...... 99 Majee Manoj ...... 137,138,351,352 Meena Rajendra Prasad ...... 216 Laha Debabrata ...... 109 Maji Ranjan Kumar ...... 259 Meena Seema ...... 235 Lakhwani Deepika ...... 205,317 Maji Sourobh ...... 192 Meena Shashi ...... 159,169 Lakra Nita ...... 122,226 Majumdar Arkajo ...... 214 Megha Swati ...... 312 Lakshmamma P...... 142,204 Majumdar Rajtilak ...... 8 Mehra Poonam ...... 263,313 Lakshmi N. Jyothi ...... 135,151 Majumder Arun Lahiri ...... 10 Mehrotra Rajesh ...... 205,209 Lal J.P...... 118 Malick N...... 43 Mehrotra Sandhya ...... 205,209 Lal Milan Kumar ...... 274 Malik Aubid ...... 223 Mekala Nageswara Rao ...... 109 Lal P...... 262 Malik Naveen ...... 353 Meshram Jayant ...... 153 Lalitha V.S.P...... 311 Mallikarjun N.M...... 327 Michael Rahul ...... 318 Lande Nilesh ...... 207 Mallikarjuna Kuruva ...... 306 Mina U...... 277 Larcher Mélanie ...... 9 Mallikarjuna M.G...... 53,274 Minhas J.S...... 126 Lata Charu ...... 125 Malpathak N.P...... 146 Minhas P.S...... 120,345 Lavania Dhruv ...... 155 Malpathak Nutan ...... 139,228 Minocha Rakesh ...... 8,26 Lavanya C...... 145 Mamrutha H.M...... 140,346 Minocha Subhash ...... 8,26 Lavekar Sailee ...... 241 Mamta ...... 48 Mishra Avinash ...... 232 Laxman R.H...... 343 Manashi Aditya ...... 227 Mishra B.K...... 118 Laxmi Ashverya .....124,186,204,280,296 Mandal Arunava ...... 246,259 Mishra Divya ...... 133 Leach Jan E...... 235 Mandal K.G...... 137,346 Mishra J.K...... 116 Lee Sung-Chul ...... 15 Mane Arun ...... 85,284 Mishra J.S...... 130
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 364 Author Index
Mishra Kumud B...... 86 Muthulakshmi S...... 164 Nitin Mukesh ...... 328 Mishra Manjari ...... 165,261 Muthusamy A...... 195 Nivedita D...... 141 Mishra Neeti Sanan...... 247 Mutum Roseeta Devi ...... 12 Nogia Panchsheela ...... 205 Mishra P.C...... 112 Naeem M...... 294 Nongpiur Ramsong ...... 183 Mishra Rukmini ...... 238 Nag T.N...... 227,228 Noor Jewel Jameeta ...... 256 Mishra Satish ...... 140 Nagar Shivani ...... 127,144 Nutan Kamlesh Kant ...... 111,333 Mishra Sefali ...... 20 Nagarajan M...... 74,230,320 Oudin Audrey ...... 9 Mishra Sonal ...... 195,229 Nagarajan V.K...... 52 Padaria Jasdeep Chatrath ...... 216 Mishra Sushma ...... 194 Nagegowda Dinesh A...... 235 Padmasree Kollipara ...... 252,253 Mishra Toshi ...... 220,344 Nagle Vinod ...... 241 Pakrasi Himadri B...... 89 Mishra Upama ...... 169 Naika Mahantesha ...... 334 Pal Madan ...... 98,148,159,169,174, Misra S.C...... 112 Naithani S.C...... 61,117,183 216,302,320 Mithöfer Axel ...... 250 Naiya Haraprasad ...... 184 Pal Shaifali ...... 235 Mithra Amitha C.R...... 112 Nakar R.N...... 72 Pal Tarun ...... 337 Mithra S.V. Amitha ...... 20 Nalli A...... 102 Palta J.A...... 95 Mitra Abhijit ...... 91,233 Nama Srilatha ...... 309 Pan Yong-Bao ...... 222 Mittal Shikha ...... 53,274 Nanda Satyabrata ...... 238 Panda Ashok ...... 181 Mittra Sarika ...... 156 Nandhitha K...... 292 Panda Binay ...... 84 Mockler Todd C...... 44 Nandi Ashis K. ...59,208,222,249,254,320 Panda S.K...... 163 Mog Babli ...... 341 Nandwal A.S...... 168,135,177,178 Pandey A...... 175 Mohammad Naseer ...... 316 Narasimha Swamy Konduri ...... 307 Pandey Amita ...... 15 Mohan Amita ...... 318 Narayan S...... 270 Pandey Ashutosh ...... 206,317 Mohanasundaram A...... 331 Narayankutty M.C...... 143 Pandey Bipin ...... 263,313 Mohanty Amitabh ...... 79 Narendra Gupta...... 126 Pandey Daya K...... 238 Mohapatra P.K...... 84,293 Nareshkumar A...... 133 Pandey Dinesh ...... 240 Mohapatra Sridev ...... 175,177 Narwade Ajay V...... 168 Pandey Garima ...... 206 Mohapatra T...... 88,197,329 Nataraja Karaba N...... 326 Pandey Girdhar K...... 15,113 Mohiudin Tabassum ...... 224 Natarajan S...... 288 Pandey Prachi ...... 247 Moin Mazahar ...... 180 Nath Onkar ...... 344 Pandey Rakesh ...... 178,233,335,338 Mondal Tapan Kumar ...... 128,212 Nath Pravendra ...... 299,301 Pandey Renu ...... 269,274,278 Morabito Domenico ...... 9 Nathawat N.S...... 120 Pandey S...... 102 Mrinalini Chawla ...... 78 Nautiyal C.S...... 182,300 Pandey S.P...... 301 Mudalkar Shalini ...... 150 Nautiyal P.C...... 147,327,343 Pandey Saurabh ...... 200 Mudgil Yashwanti ...... 129 Nayyar Harsh ...... Pandey Sona ...... 18 Muhammed Jamsheer K...... 186,204 105 Pandey Sonika ...... 232 Mukesh Nitin...... 220,344 Neeraj Kumar...... 168 Pandey Veena ...... 118 Mukherjee Abhishek ...... 10 Neeraja C.N...... 54,321 Pandey Vinod ...... 178 Mukherjee Chandan ...... 141 Negi Bhawna ...... 166 Pandiyan M...... 130 Mukherjee Gairik ...... 129,184 Negi Manisha ...... 272 Pandurangam Vijai ...... 166 Mukherjee Sritama ...... 10 Negi Neelam Prabha ...... 148 Pangaan Iris Dawn ...... 103 Mukhopadhyay Pradipto ...... 153 Ngachan Shishom Vanao ...... 349 Panwar Akanksha ...... 351,356 Mullineaux Philp ...... 32,69,123,136 Nijhawan Aashima ...... 333 Panwar Azad Singh ...... 349 Muniswamy S...... 217 Nikalje Ganesh ...... 202 Paramanantham Anjugam ...... 247 Munnam Suchandranath B...... 307 Nikam T.D...... 202,293 Parashar Renu ...... 287 Mur Luis ...... 194 Nirala Ramchiary...... 344 Paraste Kamlesh ...... 345 Murtaza Malik...... 314 Niranjana P...... 329 Pareek Ashwani ...... 11,27,111, Mushtaq Ruqia ...... 285 Nisarga K.N...... 198 114,116,1 19,121,122,136,165,183,184, Mustafiz Ananda ...... 64 Nithia S. Jannita ...... 304 188,199,202,226,261,281,314,323
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 365 Author Index
Pareek Sneh L. Singla ...... 11,27,64, Pitagi Anita ...... 334 Rai Ramakant ...... 222 111,112,116,119,121,122,136,183,184, Podile Appa Rao ...... 187 Rai Vandna ...... 20,272,312 188,199,226,261,281 Pooja B...... 196 Rai Yogita ...... 196 Parekh Nita ...... 91,233 Poolman Mark ...... 65 Raigond Pinky ...... 236 Parida Adwaita ...... 201 Poonia Vijay ...... 272 Raina Aparna Shree ...... 325 Parida Asish Kumar ...... 181 Poonia Shefali ...... 171 Raina Susheel ...... 120 Parida S.K...... 43,104,319,355 Prabhu K.V...... 112,142 Raj A...... 277 Parihar Vimala ...... 35 Pradeep T...... 114 Raj Lakshmi ...... 274 Park Se Won ...... 115 Prasad Ch. Ram ...... 135 Raj S.R. Kiran ...... 233 Parmar Hemangini ...... 297 Prasad Manoj ...... 56,206,209,244 Rajabhoj Mohit ...... 189 Parmar Rajni ...... 325 Prasad P.V.V...... 130 Rajam M.V...... 48,248,289 Parry Martin A.J...... 66 Prasad S.R...... 288 Rajan S...... 158 Partheeban C...... 130 Prasad Sai ...... 140 Rajanna M.P...... 30,197,327,328,329 Parupalli Swathi ...... 217 Prasad Subhash Chandra ...... 136 Rajeev Ranjan ...... 352 Parveen Shaista ...... 131 Prasad T.G...... 224,341 Raju B.R. .. 30,180,196,327,328,329,334 Pasala Ratnakumar ...... 276,345 Prashant Vikram ...... 318 Raju M.B...... 198 Patel C.B...... 269 Prathibha M.D...... 30,234,327,329 Rakesh K...... 146 Patel Kiran ...... 159 Pratibha M...... 196 Rakesh Verma ...... 164 Patel Manish Kumar ...... 232 Prayaga Lakshmi ...... 142,204 Ram Phool Chand ...... 156 Patel Prashanti Devendra ...... 266 Preethi N.V...... 180 Ramachandra R...... 180,328 Pathak A.D...... 155 Priyanka A.B...... 196,234 Ramakrishnan Shiv...... 144 Pathak Garima ...... 299 Pudi Vikram ...... 91,233 Ramchiary Nirala ...... 220,328 Pathak H...... 145,169,277,278 Puli Mallikarjuna Rao ...... 187 Ramegouda H.V...... 45 Pathak Ravi ...... 55 Pundir P...... 29,158,161 Ramesh Kulasekaran ...... 268 Pathak Sumya ...... 317 Punitha S...... 282 Ramesha...... 102 Pathania Shivalika ...... 57 Punyapwar Swapnil ...... 162 Ramteke S.D...... 295 Pati Pratap Kumar ...... 167 Purayannur Savithri ...... 221,234 Ramu S.V...... 45,198 Patil Amol ...... 135 Purty Ram Singh ...... 268 Rana Dheeraj ...... 140 Patil Hemant ...... 331 Purushottam...... 171 Ranawana S.R.W.M.C.J.K...... 95 Patil Mahesh ...... 224 Pushkar Suchitra ...... 172 Rane Jagadish ...... 120 Patil R.P...... 131 Pushpa D...... 196,327 Rangani Jaykumar ...... 181 Patil Supriya ...... 264 Puskar Suchitra ...... 146,296 Rangappa Krishnappa ...... 349 Patil Suyash ...... 217 Puthur Jos ...... 303 Rani P. Prasuna ...... 174 Patil Vidya ...... 146 Rachel Jacob Sherry ...... 230 Rani Y. Ashoka ...... 174 Patra Ashok ...... 268 Radadiya Nidhi ...... 302 Ranjan Aashish ...... 110 Pattiwala Yasir Umar ...... 259,341 Radha Beena ...... 143 Rao A.R...... 112,216 Paul Asosii ...... 215 Raghava Nisha ...... 237 Rao Ch. Srinivasa ...... 135 Paul Priyanka ...... 123 Raghava Ravindra Pratap ...... 237 Rao G. Lokanadha ...... 13 Paul Vijay ...... 178,233,335,338 Raghavendra A.S...... 67,187,188, Rao I. Subhakara ...... 54 Pazhamala Lekha ...... 216,217 229,230,232,305,306 Rao K.L. Narasimha ...... 174 Pednekar Suhas ...... 241 Raghothama K.G...... 52 Rao K.V...... 180 Penna Suprasanna ...... 202 Raghuram Nandula ...... 55,191 Rao M.V...... 146,157 Peter Wenzl...... 318 Raghuvanshi Saurabh ...... 12 Rao P.R...... 63,321 Petla Bhanu ...... 351 Raghuvanshi Utkarsh ...... 12 Rao S.S...... 130 Phazang Paomipem ...... 123 Raghuveer Rao Puskur...... 307 Rao Sombir ...... 215 Phukan Ujjal J...... 229 Rai Krishan Mohan ...... 210 Rao T.G.N...... 130 Pieter Edgar ...... 250 Rai Neha ...... 178 Rao Uma ...... 251 Pillai M. Arumugam ...... 326 Rai Puja ...... 147,327,343 Ratcliffe R...... 194
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 366 Author Index
Rathanakumar A.L...... 302 Sablok Gaurav ...... 202 Sarkar Deepti ...... 259 Rathi Divya ...... 152 Sacaria Vinod ...... 176 Sarkar Poulami ...... 237 Rathi Iti ...... 287 Sadashiva A.T...... 343 Sarla N...... 307 Rathod Khyati J...... 42 Saeed Bushra ...... 223 Sarma Anjan Kumar ...... 349 Rathod T.H...... 152 Saha Chinmay ...... 184 Sarma C.M...... 176 Rathor Ray Singh ...... 116 Saha Jayeeta ...... 225 Saroj P.L...... 341 Rathore Abhishek ...... 322,323 Saha Sudip ...... 33 Saroop Shveta ...... 107 Rathore M...... 345 Saha Sunayan...... 345 Sarpras M...... 220,328,344 Rathore Ray Singh ...... 11 Sahi Shivendra ...... 272 Sastry P.S...... 231 Rathore S.S...... 170 Sahoo K.K...... 27,64,121, 281, 289 Satdive Ramesh ...... 139 Rathore V.S...... 120 Sahoo L...... 172 Sathish P...... 135,151 Raveendran M...... 163,164,329 Sahoo R.N...... 92,118,335,338,343 Satyamoorthy K...... 195 Ravi Iyyakutty ...... 333 Sahu Alok Kumar ...... 183 Satyavathi C. Tara ...... 216 Ravichandran V...... 154,308 Sahu Balram ...... 117 Savita ...... 349 Ravikanth G...... 114 Sahu Pranav Pankaj ...... 244 Sawant S.V...... 210 Ravikiran K.T...... 173 Sahu Sarika ...... 216 Saxena A...... 348 Ravindran Nevedha ...... 189 Sai Prasad S.V...... 287 Saxena D.C...... 140,287 Rawal S...... 126 Sailaja B...... 307,339,349 Saxena Rachit K...... 217,322 Raxwal Vivek Kumar ...... 176 Sain R.S...... 126,273,357 Saxena S.N...... 170 Ray Krishna ...... 141 Saini Niveditha ...... 112 Schaaf Gabriel ...... 109 Raychaudhuri S...... 346 Sairam R.K...... 142,275 Scheel Dierk ...... 31 Raza S.K...... 242 Sajeevan R.S...... 326 Scheffler Brian ...... 199 Reddy A.R.150,151,180,231,307, 303,308 Sakai Hajime ...... 79 Schnell Danny ...... 37 Reddy C.S...... 264 Saketi Ananda Rao...... 314 Scholz Sandra ...... 250 Reddy Chandrashekar ...... 224 Sakhare Akshay ...... 142,147,327 Seal Anindita ...... 129,184 Reddy D. Damodar ...... 267 Salini K...... 135 Sekhar K.M...... 308 Reddy Kanubothula Sita Rami...... 151 Salvi Prafull ...... 138,352 Sekhar Sudhanshu ...... 247 Reddy M.K...77,129,200,225,239,240,315 Samanta Subhasis ...... 213 Sekhwal Manoj Kumar ...... 233 Reddy V. Amarnatha ...... 133 Sameer C...... 217 Selter Liselotte ...... 243 Reddy V.R...... 97 Samundeswari R...... 163,164 Semwal D.P...... 156 Reddy Vijay ...... 240,315 Sanagala Raghavendrarao ...... 272 Semwal Vimal ...... 134 Reema Khurana...... 352 Sane A...... 331 Sen Soumitra Kumar ...... 150 Rey Soham ...... 22 Sane A.P...... 299,300,301 Sen Sunetra ...... 177 Rice Elena...... 76 Sane P.V...... 331 Senapati Dhirodatta ...... 13 Richter Sandra ...... 201 Sane V.A...... 182,299,300 Sengupta Sonali ...... 10 Robertson Masumi ...... 81 Sangita Sushree ...... 203 Senthil A...... 282 Robin S...... 197,308,329 Sangwan Chetna ...... 209 Seth Romit ...... 325 Roopendra Kriti ...... 222 Sangwan Rajender Singh ...... 38 Shaanker R. Uma ...... 114 Roorkiwal Manish ...... 215,322,323 Sanyal Sibaji K...... 15,113 Shah Diti ...... 177 Rout Ellojita ...... 238 Sarada C...... 204 Shah Divya ...... 98,159 Roy Amit ...... 246 Saradhi A.V. Pardha ...... 102 Shamim M.Z...... 175 Roy Shweta ...... 222,249 Sarala N...... 197,329 Shamkuwar G.R...... 152 Roy Suchismita ...... 119,314 Saraswati Nayar ...... 78 Shankar Rama ...... 211 Roy Suman ...... 132 Sarathambal C...... 345 Shankar Ravi ...... 57 Roy Tapas K...... 106 Sardar Atish ...... 252 Shanker Arun K...... 135 Ruhil Kamal ...... 309 Sarin Neera Bhalla ...... 123,136,148,253 Shankhdha Shailesh Chandra ...... 263 Ryan Elizabeth P...... 235 Sarkar Arijita ...... 259 Shankhdhar Deepti...... 127,128,263 Rydquist Shane W...... 218 Sarkar B...... 135,151 Shankhdhar S.C...... 127,128
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 367 Author Index
Shanthi N...... 304 Shashibhushan N.B...... 334 Singh Anil ...... 210 Sharan Ashutosh ...... 183,184 Sheeba...... 309 Singh Anil Kumar ...... 57 Sharma Abhishek ...... 271 Sheeba J. Annie ...... 153 Singh Anoop ...... 220 Sharma Amrapali ...... 201 Shekhar Shubhendu ...... 133 Singh Anshu ...... 155,262,271 Sharma Amrita ...... 271 Shekharappa Gurumurthy ...... 143 Singh Anuj ...... 156 Sharma Arun ...... 201 Shembekar W.K...... 152 Singh Anup Kumar ...... 235 Sharma Chhavi ...... 289 Sheoran Promila ...... 220 Singh Anupriya ...... 249 Sharma D.K...... 29,158,161 Sheoran Sonia ...... 140 Singh Ashok K...... 20 Sharma Deepika ...... 127,128,205 Sheri Vijay ...... 77,225 Singh Atar ...... 178,233,335,338 Sharma Eshan ...... 186 Sherp Ashley ...... 58 Singh Balwant ...... 20 Sharma Gaurav ...... 233 Sheshshayee M.S...... 30,180,196,197, Singh Bharti ...... 286 Sharma Indu ...... 140,346 234,327,328,329 Singh Bhupinder ...... 270 Sharma Isha ...... 167 Shine M.B...... 178 Singh Brajesh ...... 126,236 Sharma Kamal Dev...... 105 Shiriga Kaliyugam ...... 53,274 Singh Brinderjit ...... 138 Sharma Kamal Dutt ...... 261 Shirokov Aleksandr...... 60 Singh D.K...... 102 Sharma Kewal K...... 331 Shivaraj S.M...... 332 Singh D.V...... 296 Sharma Laxmi ...... 282 Shivashankara K.S...... 106 Singh Deepika...... 300 Sharma Madhu ...... 289 Shome S...... 163 Singh Deepjyoti ...... 208 Sharma Mamta ...... 217 Shrivastava Madhup ...... 276 Singh Dhriti ...... 186 Sharma Manoj K...... 203,314,342 Shrivastava Sanya ...... 16,146,189,191 Singh Dipali ...... 65 Sharma Manvi ...... 204 Shrivastva Ruchi Nag ...... 227 Singh G.P...... 112,178 Sharma Meenakshi...... 250 Shukla Alok ...... 118,263 Singh Gagandeep ...... 210,221,325 Sharma Mohan A...... 124 Shukla Brihaspati N...... 124 Singh Geeta ...... 181 Sharma Namisha ...... 244 Shukla Neha ...... 51 Singh Harpreet ...... 114 Sharma Namrata ...... 355 Shukla Rakesh K...... 229 Singh Ishwar ...... 174,297 Sharma Naveen ...... 219 Shukla S.K...... 262,271 Singh Jagdev ...... 261 Sharma P.C...... 158,161 Shukla Tapsi ...... 205 Singh Jagdish ...... 143 Sharma P.K...... 167,310 Shukla Tushita ...... 286,287 Singh Jogendra ...... 29,161 Sharma Parbodh C...... 29,161,165,173 Shukla Yogesh M...... 288 Singh Karmal ...... 261 Sharma Prakash C...... 28 Siddappa Sundaresha ...... 189 Singh Khangembam ...... 213 Sharma R...... 283,348 Siddiqi E.A...... 180 Singh Kushal ...... 332 Sharma R.P...... 197,329 Siddiqi I...... 102 Singh Manjul ...... 280,296 Sharma Ram Kumar ...... 210,221,325 Siddique K.H.M...... 95 Singh Manoj Kumar ...... 201 Sharma Rameshwar Prasad ...... 3 Sidhu Gurpreet Kaur ...... 205 Singh Mohan ...... 272 Sharma Rinku ...... 53,274 Sikdar Samir Ranjan ...... 237 Singh Mohan B...... 73,80 Sharma Rita ...... 203,314,339,342 Silas Wungrampha ...... 199 Singh Muneendra ...... 323 Sharma Sadan ...... 208 Silvester Naveen ...... 90 Singh N.K...... 112 Sharma Shivam ...... 298 Sindhuja Vajravel...... 291 Singh N.P...... 143 Sharma Shweta ...... 64 Sindhumol P...... 143 Singh Nagendra K...... 20 Sharma Surinder Kumar ...... 170 Singal Rajesh Kumar ...... 197,329 Singh Name ...... 126 Sharma T.R...... 22,330 Singh A...... 102 Singh Namita ...... 337 Sharma Vasundhara ...... 270 Singh A.K...... 43,74,197,273,320,329 Singh Narendra ...... 322 Sharma Veerendra Kumar ...... 245 Singh A.L...... 72,159,269 Singh Narpinder ...... 123 Sharma Vijay K...... 76 Singh Ajay ...... 120 Singh Navneet ...... 191 Sharma Vinay ...... 148 Singh Amar Pal ...... 300 Singh Neer ...... 319 Sharma Vineet ...... 220,328,344 Singh Amarjeet ...... 15 Singh Neera ...... 127,144,347 Sharma Vishakha ...... 16 Singh Amit Kumar ...... 165 Singh Nidhi ...... 249,254 Shasany Ajit Kumar ...... 235 Singh Anandita ...... 332 Singh P.K...... 112
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 368 Author Index
Singh Pawan Kumar ...... 318 Somasundaram G...... 288 Surekha K...... 54 Singh Prabhjeet ...... 114,138 Soni Dharamvir ...... 178 Surendra S...... 102 Singh Preeti ...... 166 Soni Praveen ...... 183,184,188 Suryanarayana V...... 217 Singh Priya ...... 300 Soni Sweta ...... 220 Sutradhar Tapan...... 141 Singh R...... 345 Sonti Ramesh V...... 46 Sutton Tim ...... 323 Singh R.K...... 155,262,271 Sood Hemant ...... 298 Swain P...... 343 Singh Rakesh ...... 165 Sopory S.K...... 64,112,121 Swamy K.N...... 321 Singh Rakesh K...... 103 Soumen Bhattacharjee ...... 140,141,227 Swapna M...... 155 Singh Ravindra Pal...... 357 Soumya P.R...... 302 Swarnalakshmi K...... 272 Singh Renu ...... 278 Sowmya H.R...... 197,329 Swarnkar Mohit Kumar ...... 57, 210 Singh S...... 116 Spall Kaur Nirmal K...... 179 Swaroop Vanka ...... 252 Singh S.K...... 116 Sreedeep S...... 172 Swathy P.S...... 195 Singh S.P...... 155,262,271 Sreeharsha R.V...... 307 Syiemlieh Hadienlarisa ...... 349 Singh Sandeep ...... 266 Sreevathsa Rohini ...... 334 Tahir Inayatullah ...... 83 Singh Shardendu K...... 97 Sri Tanu ...... 332 Taj Gohar ...... 240 Singh Shreenivas K...... 234 Srikanth B...... 54 Talwar H.S...... 101,130 Singh Smita ...... 155,262,271 Srimany Amitava ...... 114 Tanushri Kaul ...... 314 Singh Sudhanshu ...... 156 Srinivas Agurla ...... 188 Tasleem Mohd ...... 240 Singh Sukhbir ...... 149 Srinivasan Samineni ...... 215,323 Tavva Sresty ...... 79 Singh Sukhwinder ...... 318 Sritharan N...... 348 Teja V. Dharma ...... 233 Singh Sunil Kumar ...... 210 Srivani V.P...... 233 Tetali Sarada ...... 232 Singh Supreet ...... 114 Srivastava Anjil Kumar ...... 13 Tewari S...... 163 Singh Surendra Pratap ...... 210 Srivastava Ashish Kumar ...... 156,202 Thakur A.K...... 137,346 Singh V.K...... 158 Srivastava J.P...... 118 Thakur Jitendra K...... 192,211,213,338 Singh V.P...... 127,132,144,286,347 Srivastava Jayanti ...... 237 Thamilarasi K...... 331 Singh Vijayata ...... 208 Srivastava M.K ...... 271 Thawait Lokesh Kumar ...... 42 Singh Vijendra ...... 165 Srivastava Monika ...... 278 Thayssa Schley...... 304 Singh Vikas K...... 217 Srivastava S...... 171 Thirunavukkarasu Nepolean ...... 53,274 Singh Vikash ...... 212 Srivastava Shruti ...... 301 Thiruvettai Vivek ...... 326 Singh Vinay...... 342 Srivastava Smriti ...... 299 Thomas Payne ...... 318 Singh Yogeshwar ...... 345 Srivastava V.K...... 155 Thomas S...... 171 Singla Priyanka ...... 125 Srivastava Vikas ...... 221,234 Thombare Nandkishore ...... 331 Sinha Alok Krishna ...... 14,185,244 Steinhorst Leonie ...... 15 Thudi Mahendar ...... 215 Sinha Neelima ...... 110 Stephen Roy ...... 226 Thulasiram H.V...... 234 Sinha Pallavi ...... 217 Stierhof York ...... 201 Tikaniya Divya ...... 220 Sinha Sangram ...... 356 Strader Lucia ...... 58 Tikkanen Mikko ...... 109 Sinha Somya ...... 176 Subbarayaudu B...... 130 Tittal Megha ...... 276 Sinha Veena ...... 247 Subbiah V...... 102 Tiwari Archana ...... 181 Sinharoy Senjuti ...... 199 Subrahmanyam D.... 54,63,174,307,339, 349 Tiwari Manish ...... 316 Sirhindi Geetika ...... 179,285 Subramaniam V.R...... 331 Tiwari Rahul ...... 92,172,343 Sivakumar R...... 154,292 Subramanyam R...... 309 Tiwari Shalini ...... 125 Sivaprakasam J.V...... 295 Suchendranathbabu M...... 321 Tiwari Sneha ...... 190 Sivaraju K...... 124 Sudhakar Chinta ...... 133,254 Tiwari Sushma ...... 20 Skottke Kyle ...... 90 Sujatha T...... 311 Tokas Indu ...... 15 Slewinski Tom ...... 76 Sumbria Veena ...... 355 Tomar Ajeet Singh ...... 280 Smitharani J...... 327 Sunder Singh ...... 168 Tomar Vandana ...... 205 Smitharani J.A...... 329 Suneja Yadhu ...... 149 Townsley Brad ...... 110 Snedden W.A...... 114 Sunil Bobba ...... 67 Tracy Lawson ...... 69
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 369 Author Index
Tripathi A.K...... 27,136,281 Vashist Enesh...... 298 Wakchaure G.C...... 276 Tripathi N...... 345 Vemanna Ramu S...... 254 Wang Jun ...... 322 Tripathi Shailesh ...... 104,323 Venkatesh B...... 133 Wani Zahoor ...... 224 Tripathy B.C...... 68,309 Vennapusa Amaranatha ...... 254 Wany Aakanksha ...... 195 Tripathy Jayant ...... 199 Vergish Satyam ...... 162,193 Watt Mitchelle ...... 140 Trivedi A.K...... 160 Verma Baburam ...... 77,239,240 Weiming He...... 322 Trivedi P.K...... 40,205,206,317,318 Verma Deepanjali ...... 185 Weinig Cynthia ...... 44 Trivedi S.M...... 145 Verma Indu ...... 222 Westall Corey ...... 58 Tula Suresh ...... 118 Verma Monika ...... 220 Wicker Thomas ...... 243 Turlapati Swathi ...... 8,26 Verma Pooja ...... 137 Wong Chui E...... 80 Tuteja Narendra ...... 118 Verma Praveen K...... 50,221,247,234,258 Woody Scott T...... 44 Tuteja Omprakash ...... 140 Verma Rakesh ...... 16,157,171,172, 191, Xie Qiguang ...... 44 Tyagi A.K...... 43,153,104,250,352, 282,296,324,343 Yadav A.K...... 15,113 353,355 Verma S.K...... 160 Yadav Amita ...... 209 Tyagi Isha ...... 245 Verma Sandhya...... 258 Yadav Archana ...... 145,220,338 Tyagi Shikha ...... 332 Verma Subodh ...... 200 Yadav Chandrabhan ...... 206 Udayakumar M...... 30,45,180,196,198, Verma Susheel ...... 354 Yadav D.K...... 115 224,234,254,327,329,334 Verma Vibha ...... 350 Yadav Gitanjali ...... 93,336,338 Uddin Moin ...... 291,294,298 Veronica N...... 174 Yadav Karuna ...... 267 Udhayakumar Nagan ...... 291 Vetcha Aswani...... 187 Yadav Mahesh ...... 333 Udvardi Michael ...... 199 Vigneswaran Veena ...... 143 Yadav Mohit ...... 220 Ulrike Bechtold ...... 69 Vijay Priyanka ...... 112 Yadav N.D...... 120 Umesh M.R...... 131 Vijaya Bramha...... 344 Yadav P...... 171 Upadhyay Natvarlal V...... 288 Vijayakumar H.P...... 288 Yadav Pallavi ...... 228 Upadhyay Rakesh ...... 301 Vijayalakshmi D...... 163,164 Yadav Praduman ...... 145 Upadhyaya C.P...... 115,262,281 Vijayalakshmi P...... 321 Yadav Pragya ...... 164,191 Upadhyaya D.C...... 262 Vijayaraghavan H...... 130 Yadav Rachita ...... 51 Upadhyaya H...... 163 Vijaybhaskar V...... 102 Yadav Rakesh ...... 337 Upadhyaya H.D...... 104 Viji Maluventhen ...... 291 Yadav S...... 171 Upadhyaya Hari Deo ...... 272 Vimalakumari N.K...... 255 Yadav S.K...... 135,151 Vadassery Jyothilakshmi ...... 250 Vineeth T.V...... 165 Yadav Sanjay ...... 148 Vadez V...... 101 Vinod K.K...... 74 Yadav Saurabh ...... 94,335 Vadez Vincent ...... 313 Vinod Kumar ...... 278 Yadav Shashank K ...... 16,146,157,164, Vaganan M. Mayil ...... 333 Vinya V...... 102 172,189,191,324 Vagheera P...... 151 Virdi Singh ...... 123 Yadav Sonam ...... 220 Vaishnav Piyush R...... 288 Vishnukiran T...... 321 Yadav Sudesh Kumar ...... 117 Vaishnaw Vivek ...... 316 Vishwakarma C...... 171,172,327 Yadava Pranjal ...... 174,297 Vakharia Dinesh N...... 255 Vishwakarma Harinder ...... 216 Yadukrishnan P...... 166 Vanaja M...... 135,151 Vispo Naireen A...... 103 Yamini K.N...... 217 Vanvari Rythm ...... 195 Viswanathan C...... 16,92,146,147,157, Yasin Jeshima Khan ...... 326 Varsani Suresh ...... 47 164,171,172,189,191,216,230,275,282, Yogindran Sneha ...... 248 Varsheny Rajeev K...... 322 296,324,326,327,341,343 Yongle Li ...... 323 Varshney Lalit ...... 291,298 Voleti S.R...... 54,63,143,174,307,321, Zala P.V...... 159,269 Varshney Rajeev K...... 215,216,217,323 339,349 Zhang Gengyun ...... 322 Vashisht Ira ...... 337 Vyas Dhiraj ...... 113,347 Zhou W.H...... 172 Zinta Gaurav ...... 110
IPPCongress 2015 3rd International Plant Physiology Congress “Challenges and Strategies in Plant Biology Research” December 11-14, 2015, JNU, New Delhi, India 370