Phenetic Characterization, Molecular Phylogeny and Bio-Prospecting of Selected Saline Microalgae from Indian Sub-Continent
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PHENETIC CHARACTERIZATION, MOLECULAR PHYLOGENY AND BIO-PROSPECTING OF SELECTED SALINE MICROALGAE FROM INDIAN SUB-CONTINENT Thesis submitted to the COCHIN UNIVERSITY OF SCIENCE AND TECHNOLOGY In partial fulfillment of the requirements for the Degree of Doctor of Philosophy in Marine Biology Under the Faculty of Marine Sciences By PREETHA. K (Reg. No. 3780) Marine Biotechnology Division Central Marine Fisheries Research Institute Post Box No. 1603, Ernakulam North P.O., Kochi-682 018 October 2015 भा.कृ .अनु.प.- के द्रीय खारा जललजीवपालनअनुसंधानसंथान ICAR-Central Institute of Brackishwater Aquaculture #75, संथोमहाई रोड़ राजा अणामलै पुरम चेनई – 600 028, तिमलनाडु, भारत (ISO 9001:2008 certified) Indian Council of Agricultural Research, Ministry of Agriculture, Govt. of India 75, Santhome High Road, R A Puram, Chennai 600 028 Tamil Nadu, India डॉके .के .िवजयन, पी.एच.डी., ए.आर.एस, िनदेशक Dr. K.K. Vijayan, Ph. D., ARS, Director This is to certify that the thesis entitled “Phenetic characterization, molecular phylogeny and bio-prospecting of selected saline microalgae from Indian sub-continent” is a bonafide record of research work carried out by Ms. PREETHA. K (Reg. No. 3780) under my supervision at Central Marine Fisheries Research Institute (CMFRI), Kochi for partial fulfillment of requirement for the degree of Doctor of Philosophy in Marine Sciences, Cochin University of Science and Technology, Kochi, and that no part of this work has previously formed the basis for the award of any degree, diploma, associate-ship, fellowship or any other similar title or recognition in any universities or institutes. Dr. K K Vijayan October 2015 (Supervising Guide) Kochi-18 (Former Principal Scientist and Head, Marine Biotechnology Division Central Marine Fisheries Research Institute Kochi-682018) I hereby do declare that the thesis entitled “Phenetic characterization, molecular phylogeny and bio-prospecting of selected saline microalgae from Indian sub-continent”, is an authentic record of research work done by me under the supervision of Dr. K. K. Vijayan, Director Central Institute of Brackish water Aquaculture, Chennai, (former Principal Scientist & Head, Marine Biotechnology Division, CMFRI, Kochi, 682018) and that no part of this work, has previously formed the basis for the award of any degree, diploma associate-ship, fellowship or any other similar title of any university or institution. Kochi-18 Preetha K October 2015 (Reg. No. 3780) I have been fortunate to have Dr. K. K. Vijayan, Director, CIBA, Chennai (former Principal Scientist and Head of Marine Biotechnology Division, CMFRI, Kochi), as my mentor and supervising guide, who has inspired me to think further than I thought I could. I will be grateful to him forever for his flawless support and encouragement throughout the work. PhD was always a dream, and it came through the CSIR-UGC JRF. Here, I acknowledge UGC for their financial support provided for those five years. I also acknowledge National Agriculture Innovation Project (NAIP), ICAR, for the financial support provided for developing the culture collection. Being a part of CMFRI was a memorable and rewarding experience and it would have been not possible unless Dr. A. Gopalakrishnan, Director and Dr. G. Syda Rao, Former Director, CMFRI and had given me a chance. I extend my thanks to them from the bottom of my heart. Dr. P.C. Thomas, Principal Scientist (Retd.), was a part of our division and was the SIC of HR division. His timely decisions and interventions made with funding agency and university could not be forgotten. I will be indebted to him for all his support. I express my sincere gratitude to the Head of Marine Biotechnology Division Dr. P. Vijayagoplal, Principal Scientist for his value adding suggestions and the support provided throughout the period of my work. I thank all the scientists of the division for their unlimited support and motivation. Particularly want to acknowledge Mr. N.K. Sanil, for his help provided during microscopic examinations and proof reading of the write up and Dr. Kajal Chakraborty, for his assistance given during chemical evaluation. I thank Dr. Srinivasa Raghavan V., Dr. Pradeep M. A., Dr. Sandhya Sukumaran, and Dr. C. P. Suja and other scientists of the division. I specially render my gratitude to the technical staffs, Mr. K. Surendran for his valuable suggestions, Mr. S. Nandakumar Rao for helping out me in doing the nutritional profiling and Dr. M. P. Paulton, for rendering helping hands during molecular analysis. I also thank Mrs. P. Vineetha, Mrs. Parukkutty Mr. Gireesh P. M., Mr. Shaji A. K., and Mr. Baiju for their limitless help rendered during different phases of my PhD. I am grateful to Dr. K.J. Joseph, Professor (Retd.), Department of Marine Biology, Microbiology and Biochemistry, CUSAT for his guidance during initial period of my work, particularly in identification of microalgae. I wish to express my gratitude to Dr. P. Kaladharan, Principal Scientist, CMFRI who provided timely support during my sample collections and taxonomic identification. I extend my special thanks to Dr. T. V. Sathianandan, Principal Scientist and Head, FRAD, CMFRI and Dr. Jayalakshmi, Chief Scientist, NIO, Kochi for their critical advice rendered in statistical analysis of my data. I would like to thank Dr. Rita Jayashankar and Dr. Shoji Joseph (Principal Scientists, CMFRI) for sharing algal culturing facilities. I sincerely thank scientists of CMFRI - Dr. Joe K. Kizhakudan, Dr. P.K. Asokan, Dr. Sunil Mohammed, Dr. V. Kripa, Dr. D. Prema, Dr. Shyam S. Salim, Dr. Rema Madhu Dr. Madhu K. and Dr. K. S. Sobhana for their support and cooperation provided during my work. I also thank other staffs of all scientific divisions, accounts and audit, bill & cash, library, central lab, canteen etc. Like to mention the names of Mr. Khambadkar (FRAD), Ms. K. P. Salini (ATIC), Ms. G Shylaja (central lab), Mr. A. Padmanabha (E&M) and Mr. K. M. Venugopalan for their prompt help during various stages of my work. I thank wholeheartedly, Dr. Bobby Ignatious (SIC), Mr. C N Chandrasekharan and other staffs of HRD for all their assistance, including taking timely steps in forwarding the files and other documents. I would like to express my deep sense of gratitude to the Doctoral and Research Committee members faculty members of Department of Marine Biology, Microbiology, Biochemistry & School of Industrial Fisheries, CUSAT including Dr. Aneykutty Joseph, my external subject expert, Dr. Rosamma Philip & Dr. M. Harikrishnan, my examiners, and Dr. Mohd Hatha for their support and helpful suggestions. I sincerely thank Dr. C. N. Ravishankar, Director, CIFT, Dr. Zynudheen A.A., Senior Scientist, CIFT, and Dr. T. V. Shankar, Principal Scientist, CIFT for their helpful ideas. I appreciatively acknowledge Dr. V. Sivasubramanian, Director, Vivekananda Institute of Algal Technology (VIAT), Chennai, Dr. L. Uma, Director, NFMC and Professor, Bharathidasan University, Thrichy and Dr. Malliga P., Professor, BDU, for their valuable suggestions. My co-scholars are my strength, with whom I shared my thoughts and my feelings, from whom I learned the laboratory techniques and important lessons of Biotechnology, and who extended their hands to help me unlimitedly. My dear friends - members of “TEAM MBTD” thank you all. Specially want to acknowledge Dr. Lijo John (present Asst. Director, Export Inspection Council) and Subin C. S. who helped me to initiate the work. I express my deep sense of gratitude to Dr. Neil Scolastin, and Dr. P.A. Vikas who were there to support throghout. Also thank Mrs. Anusree. V. Nair, Sandhya S.V., Reynold Peter, Dr. Praveen N. K, Dr. Deepu Joseph, Dr. Sajesh S. V., Dr. Jeswin Joseph, Dr. Anju Antony, Mr. Leo Antony, Mrs. Esha Arshad, Mr. Arunkumar T. V., Dr. Meera Menon, Mrs. Suja Gangadharan, Mr. Renjith Kumar, Mr. Iyyapparaja, Mrs. Jaseera Sabeel, Mr. Sayooj P., Mrs. Selsa J. Chakkalakkal, Mrs. Bini Thilakan, Mr. Binesh C.P, Mr. Shamal P., Mr. Jijo, Dr. Sajeela K. A, Vidhya K Ms. Sheethal and Mrs. Githa (Mariculture division), Mrs. Preetha G. Nair (FEM Division). I also thank all research scholars and skilled staffs of CMFRI for their helping hands. I expand my gratitude to the cleaning staffs and the security guards who maintain clean and safe surroundings. Once again I thank all those who lend a hand during my PhD work. Without support from my family I would have not completed this journey of PhD. My parents always stood by me, and supported me unconditionally. I am obliged to them beyond words for their affection and blessings. My sister Nandini, always was a patient companion with whom I shared my anxieties and stresses during my work. I express my sincere love and gratitude to her. I also thank my in-laws for their prayers and blessings. I am fortunate to have a perfect life partner Suresh, my husband, who was with me at each step of my work, as a supporter and an adviser. I express my respect and appreciation to him. My dear boy, Sumesh – from his limits he always encouraged and supported me. Thank you my son for being so adoring. Above all I thank the Divine Power, for showering blessings on me throughout my life and keeping me always happy and contented. Preetha K 1. Introduction and Scope of Thesis --------------------------------------------------------- 1 1.1 What are Microalgae? --------------------------------------------------------------------- 1 1.2 Ecological Perspective of Microalgae --------------------------------------------------- 3 1.3 Molecular Taxonomy of Microalgae - an Emerging Field: