A Method of Multiplex PCR for Detection of Field

Total Page:16

File Type:pdf, Size:1020Kb

A Method of Multiplex PCR for Detection of Field World J Microbiol Biotechnol (2015) 31:675–679 DOI 10.1007/s11274-015-1821-6 SHORT COMMUNICATION A method of multiplex PCR for detection of field released Beauveria bassiana, a fungal entomopathogen applied for pest management in jute (Corchorus olitorius) Chinmay Biswas • Piyali Dey • B. S. Gotyal • Subrata Satpathy Received: 9 December 2014 / Accepted: 7 February 2015 / Published online: 14 February 2015 Ó Springer Science+Business Media Dordrecht 2015 Abstract The fungal entomopathogen Beauveria bassi- jewellery and home decorations (Kundu 1956). Jute is of ana is a promising biocontrol agent for many pests. Some two kinds namely, tossa jute (Corchorus olitorius) and B. bassiana strains have been found effective against jute white jute (C. capsularis). Jute is severely affected by pests. To monitor the survival of field released B. bassiana various insect-pests such stem weevil (Apion corchori), a rapid and efficient detection technique is essential. jute semilooper (Anomis sabulifera), Bihar hairy caterpillar Conventional methods such as plating method or direct (Spilosoma obliqua) etc. (Ramasubramanian et al. 2010). culture method which are based on cultivation on selective Earlier reports suggest that jute pests particularly Bihar media followed by microscopy are time consuming and not hairy caterpillar and semilooper can be controlled by so sensitive. PCR based methods are rapid, sensitive and Beauveria bassiana (Pandit and Som 1988). Apart from reliable. A single primer PCR may fail to amplify some of causing ecological hazards complete reliance on chemical the strains. However, multiplex PCR increases the possi- pesticides becomes a burden on resource poor jute farmers bility of detection as it uses multiple primers. Therefore, in of the Indian subcontinent. Therefore, efforts are on to the present investigation a multiplex PCR protocol was substitute some use of pesticides by low cost biocontrol developed by multiplexing three primers SCA 14, SCA 15 agents. B. bassiana (Balsamo) Vuillemin (Ascomycota: and SCB 9 to detect field released B. bassiana strains from Hypocreales) is an important entomopathogenic fungus. soil as well as foliage of jute field. Using our multiplex Since long it is known to cause white muscardine disease in PCR protocol all the five B. bassiana strains could be de- silkworm (Bombyx mori). Spores of B. bassiana germinate tected from soil and three strains viz., ITCC 6063, ITCC and grow directly through the cuticle to the inner body of 4563 and ITCC 4796 could be detected even from the crop their host, proliferate throughout the insect’s body and foliage after 45 days of spray. eventually kill it. It has been found useful to control many crop pests such as stem borer (Chilo partellus) in maize Keywords Multiplex PCR Á Beauveria bassiana Á Jute Á and sorghum (Maniania 1993; Reddy et al. 2009), leaf SCAR marker Á Endophyte roller (Sylepta derogata) in cotton (Ramesh et al. 1999); beetle (Leptinotarsa decemlineata) in potato (Wraight and Jute is an important fibre crop only second to cotton which Ramos 2002), aphids (Aphis spp.) as well as mites (Am- is mainly grown in the South East Asian countries like blyomma maculatum and A. americanum) in wheat (Hat- India, Bangladesh, Nepal, China, Indonesia, Thailand, ting et al. 2004; Kirkland et al. 2004) etc. Presently many Myanmar and few South American countries. It is used in commercial formulations of B. bassiana are available in making sacks, ropes, bags, carpets, shoes, geo-textiles, the market and its use is increasing. However, as it is a live product and is introduced into a particular ecosystem from outside its survival is always challenged by the native C. Biswas (&) Á P. Dey Á B. S. Gotyal Á S. Satpathy microbial populations. Therefore, monitoring of field re- Central Research Institute for Jute and Allied Fibres (CRIJAF), leased B. bassiana is essential and it requires an efficient Barrackpore, Kolkata 700120, West Bengal, India e-mail: [email protected]; and robust detection technique. Like other fungi, B. [email protected] bassiana is conventionally detected by plating or culturing 123 676 World J Microbiol Biotechnol (2015) 31:675–679 on selective media. But these cultivation methods are time K were added and kept at 37 °C for half an hour and 0.6 consuming and not so sensitive (Biswas et al. 2012). volume of ice cold isopropanol was added. The precipitate However, PCR based methods are rapid, sensitive and re- was again centrifuged at 80009g and the pellet was washed liable in microbial diagnostics (Yamamoto 2002; Baric and with 70 % ethanol and dried at room temperature. Then the Dalla-Via 2004). The technique is recently being applied in DNA pellet was dissolved in TE buffer (10 mmol-1 Tris– detection of B. bassiana (Castrillo et al. 2003; Quesada- HCl, 1.0 mmol-1 EDTA) and stored at -20 °C. Moraga et al. 2006; Ownley et al. 2008). But, a single Genomic DNA was extracted from the young leaves of primer PCR may fail to amplify some of the strains. The both B. bassiana treated as well as untreated jute (C. oli- probability of detection in PCR can be increased by using torius) plant samples collected from the field. 300 mg tis- multiple primers in a single reaction, commonly known as sue was taken from each sample and was surface sterilized multiplex PCR (Loncaric et al. 2008). As many strains of by treating with 0.5 % sodium hypochlorite for 2 min and B. bassiana are used in pest management, multiplex PCR then with 70 % ethyl alcohol for 2 min. Then it was would be more useful for their detection. Therefore, in the thoroughly washed with sterile distilled water. DNA was present investigation a multiplex PCR protocol was de- extracted by using cetyl trimethyl ammonium bromide veloped to detect field released B. bassiana strains from (CTAB) method (Biswas et al. 2012). The sample was soil as well as foliage. crushed properly by adding Polyvinylpyrrolidone (PVP) Beauveria bassiana isolates viz., ITCC 6063, ITCC which helps to dissolve the mucilage present in jute plant. 4512, ITCC 4925, ITCC 4796 and ITCC 4644 used in the The tissues were ground in CTAB and transferred to 500 ll experiment were collected from Indian Type Culture Col- 4 M NaCl which was kept at 60 °C for 1 h with occasional lection (ITCC), Division of Plant Pathology, Indian Agri- stirring. Then equal volume of dichloromethane was added cultural Research Institute, New Delhi. All the fungal and centrifuged at 14,0009g for 15 min. The supernatant cultures were maintained at 25 °C on potato dextrose agar was carefully taken and 5 ll RNAse (5 mg/ml) was added (PDA). The leading C. olitorius variety JRO 524 was used and kept at 37 °C for half an hour. After that equal volume for the field experiments. Seeds were collected from Crop of dichloromethane was added followed by centrifugation Improvement Division, Central Research Institute for Jute at 10,0009g for 15 min. The supernatant was carefully and Allied Fibres, Barrackpore, India. taken out and was precipitated with 0.6 volume of ice cold The conidia of B. bassiana were harvested by scraping isopropanol. The precipitate was again centrifuged at the surface of fungal culture with a sterile camel hair brush 14,0009g and the pellet was washed with 70 % ethanol into a 100 ml glass beaker containing 50 ml sterile distilled and dried at room temperature. Then the DNA pellet was water. The conidial suspension was prepared by mixing the dissolved in TE buffer (10 mM Tris, 1.0 mM EDTA) and solution with a magnetic stirrer for 5 min. The concentra- stored at -20 °C. tion of conidia was adjusted to the desired concentration of For isolation of DNA from B. bassiana, monoconodial 1 9 108 conidia/ml using haemocytometer and a light cultures of all the seven strains were grown in potato microscope (409 magnifications). The conidial concen- dextrose agar (PDA) broth (pH 5.5) for 7 days at tration of B. bassiana @19 108 conidia/ml was chosen 25 ± 1 °C. The mycelia were filtered through Whatman following Biswas et al. (2012). For pest management in No. 1 filter paper. An amount of 500 mg mycelia was jute conidial suspension of the five B. bassiana isolates ground in liquid nitrogen and transferred to DNA extrac- were sprayed in the field where the crop was sown in tion buffer (100 mM Tris, 1.4 M NaCl, 20.0 mM EDTA, 3 9 2.5 m plots with four replications for each isolate in 4 % CTAB (Murray and Thompson 1980) and incubated at randomized block design along with untreated check. The 60 °C for 1 h with occasional stirring. Equal volume of crop was grown with recommended package of practices dichloromethane was added followed by centrifugation at without using any plant protection chemical. 14,0009g for 15 min. The supernatant was carefully taken Chemical lysis buffer was prepared by using CTAB out and was precipitated with 0.6 volume of ice cold iso- method (Robe et al. 2003) with major modifications. 400 mg propanol. The precipitate was again centrifuged at soil sample was grinded with liquid nitrogen and 1 ml of 14,0009g and the pellet was washed with 70 % ethanol preheated (65 °C) chemical lysis buffer was added in the and dried at room temperature. Then the DNA pellet was sample. Extraction lysis buffer was modified by adding dissolved in TE buffer (10 mM Tris, 1.0 mM EDTA) and 10 % CTAB, 40 mM Tris, 1.4 M NaCl, 20.0 mM EDTA, stored at -20 °C. The DNA yield obtained by modified 0.2 % b-marcaptoethanol and 3 % w/v PVP (Sigma). CTAB method from different soil samples varied from 370 Sample was then incubated at 60 °C for 1 h.
Recommended publications
  • RECENT LITERATURE on LEPIDOPTERA (Under the Supervision of PETER F
    1960 .loumal of the Lepidopterists' Society 161 RECENT LITERATURE ON LEPIDOPTERA (Under the supervision of PETER F. BELLINGER) F. BIOLOGY AND IMMATURE STAGES Comstock, John Adams, "Notes on metamorphoses of the Giant Skippers (LepidopTera: Megathymimc) and (he life history of an Arizona species." Rull. southern Calif. Acad. Sri., vol.55: pp.19-27, 3 figs. 1956. Describes mature larva & pupa of Mega­ thymus evalls;. [P B.] Comstock, John Adams, "Notes on the life history of a rare Arizona sphinx moth, Xylophanes faito Walker." Bull. southerll Calif. Acad. Sci., vo1.55: pp.102-106. 5 figs. 1956. Describes mature larva & pupa; foodplant B01lvardia glaberrima. LP.B.] Comstock, John A., "Brief notes on the life histories of two Arizona geometrid moths." Bull. southem Calif. A (ad. Sci., vol. 56 : pp.99-100 1957. Describes larva & pupa of Philobia aspirata (from Black Walnut), & egg & young larva of Pero modest1ls. [P.B.] Com';tock, John Adams, "Life histories of two southern Arizona moths of the genus Caripeta." Bull. southern Calif. Ilrad. Sci., vol. 56 : pp.88-96, 4 pIs. 1957. De£cribes & figures early stages of C. hilumaria (reared on willow, probably not the normal food plant) & C. macularia (reared on oak). [Po B.] Comstock, John Adams, "Notes on the early stages of two western American moths." Bull. southern Calif. Acad. Sci., vol.56: pp.42-47, 6 figs. 1957. Describes mature larva; & pupa; of Cisthene nexa & Agriopodes viridata; both feed on the lichen Ramalina combeoides. [Po B J Comstock, John Adams, "Notes on the metamorphosis of an Agave-boring butterfly from Baja California, Mexico." Trans.
    [Show full text]
  • Noctuoidea: Erebidae: Others
    Staude et al. / Metamorphosis 27: S165–S188 S165 ____________________________________________________________________________________________________________________________ Noctuoidea: Erebidae: Others Reference/ Lepidoptera Host plant Locality rearing no. Taxon Subfamily Family Taxon Family M1148 Anoba angulilinea Anobinae Erebidae Dalbergia Fabaceae Tshukudu Game melanoxylon Reserve, Hoedspruit M998 Anoba atripuncta Anobinae Erebidae Ormocarpum Fabaceae Tshukudu Game trichocarpum Reserve, Hoedspruit Gv71 Baniana arvorum Anobinae Erebidae Elephantorrhiza Fabaceae Steenkoppies, farm, elephantina Magaliesburg 14HSS52 Baniana arvorum Anobinae Erebidae Elephantorrhiza Fabaceae Steenkoppies, farm, elephantina Magaliesburg 13HSS84 Plecoptera arctinotata Anobinae Erebidae Senegalia caffra Fabaceae Steenkoppies, farm, Magaliesburg M1020a Plecoptera flaviceps Anobinae Erebidae Dalbergia Fabaceae Casketts, farm, melanoxylon Hoedspruit M317 Bareia incidens Calpinae Erebidae Ficus lutea Moraceae Casketts, farm, (unplaced as to Hoedspruit tribe) 14HSS87 Egnasia vicaria Calpinae Erebidae Afrocanthium Rubiaceae Dlinsa Forest, (unplaced as to mundianum Eshowe tribe) 12HSS163 Exophyla multistriata Calpinae Erebidae Celtis africana Cannabaceae Golden Valley, (unplaced as to Magaliesburg tribe) M416 Exophyla multistriata Calpinae Erebidae Trema orientalis Cannabaceae Sekororo, Tzaneen (unplaced as to (Fed on Celtis tribe) africana) M743 Lacera alope Calpinae Erebidae Pterolobium Fabaceae Moholoholo Rehab (unplaced as to stellatum Centre, Hoedspruit tribe)
    [Show full text]
  • Dr. Abdul Sadat Assistant Professor of Sericulture
    Dr. Abdul Sadat Assistant Professor of Sericulture Raiganj University West Bengal India Academic Qualifications: M.Sc in Zoology Ph.D in Science (Zoology) Contact Address: Department of Sericulture, Raiganj University, PO. Raiganj, Dist. U/D 733134 Email: [email protected] and [email protected] Phone numbers: +919932341504 Research Interest: Conservation Biology and Entomology Specialization: Ecology and Animal behavior Fellowships: NA Award: GATE-2010, CSIR-UGC NET-2013 (June), CSIR-UGC NET-2013 (December) Conference/Seminar/Organisation: 1. International seminar, Department of Chemistry, RGU 2. National seminar, Department of Zoology, UGB 3. International seminar, Department of Chemistry, UGB 4. National seminar, Department of Zoology, RGU 5. Rigional Science Congress, WB Govt., WB 6. National seminar, RGU Teaching Experience: 6 years Research Experience: 4 years Research Guiding Experience: NA Ph.D. Scholars: NA Post-Doctoral – Research Associate: NA Completed Research Projects: NA Ongoing Project : NA Previous & Present Employment: 1. Assistant teacher in School (L.M.C.B, kaliyaganj, U/D,WB, India) 2. Assistant professor in College (Gangarampur College, WB, India) 3. Assistant professor in University (Raiganj University, WB, India) Administrative Experience: Nodal teacher for Minority Scholarship Financial Assistances from following Organizations for attending International Conferences in abroad: NA Publications: Books: NA Research Papers: Eleven (11) Technical Report: NA Advisory Editor: NA Special Lectures: NA Keynote Addresses: NA Country Visited: NA Editorial Board Member: NA Reviewer of the following Books and Journals: One (01) Jury Member: NA LIST OF PUBLICATIONS: RESEARCH PAPERS : National: 1. Sadat A. Bhutia D. Rai BK. Pal J. 2006. Effect of cypermethrin on cytocrom P-450 elevation in fresh water cat fish, Heteropneustes fossilis.
    [Show full text]
  • Lepidoptera Fauna of Namibia. I. Seasonal Distribution of Moths of the Koakoland (Mopane) Savanna in Ogongo, Northern Namibia
    FRAGMENTA FAUNISTICA 57 (2): 117–129, 2014 PL ISSN 0015-9301 © MUSEUM AND INSTITUTE OF ZOOLOGY PAS DOI 10.3161/00159301FF2014.57.2.117 Lepidoptera fauna of Namibia. I. Seasonal distribution of moths of the Koakoland (Mopane) Savanna in Ogongo, northern Namibia Grzegorz KOPIJ Department of Wildlife Management, University of Namibia, Katima Mulilio Campus, Private Bag 1096, Katima Mulilo, Namibia; e-mail: [email protected] Abstract: During the years 2011–2013, moths were collected in Koakoland (Mopane) Savanna in the Cuvelai Drainage System, Ovamboland, northern Namibia. In total, 77 species from 13 families have been identified. Their seasonal occurrence in this habitat was also investigated, with most species recorded in wet season between September and April, but with clear peak in February and March. The family Noctuidae was by far the most speciose (38 recorded species), followed by Crambidae (8 spp.), Sphingidae (6 spp.) and Arctiidae (4 spp.). All other families were represented by 1–3 species. For each species listed date of collection is given, and data on its global distribution. Key words: Lepidoptera, check-list, biodiversity, distribution, moths, Ovamboland INTRODUCTION According to recent quite precise estimate, there are 15 5181 species, 16 650 genera and 121 families of Lepidoptera worldwide (Pouge 2009). Lepidoptera fauna of Namibia has recently attracted attention of European entomologists. However, thorough surveys were conducted hitherto in a few areas only, such as Brandberg and Hobatere. The northern regions of the country were especially badly neglected. In southern Africa (south of Zambezi and Kunene Rivers) – 8 511 species, 2 368 genera and 89 families were recently catalogued (Vári et al.
    [Show full text]
  • Modeling Hydroprene Effects on Eggs and 5 Instar Wandering Phase Larvae of the Indianmeal Moth, Plodia Interpunctella
    MODELING HYDROPRENE EFFECTS ON EGGS AND 5th INSTAR WANDERING PHASE LARVAE OF THE INDIANMEAL MOTH, PLODIA INTERPUNCTELLA (LEPIDOPTERA: PYRALIDAE) by SIVAKUMAR MOHANDASS B.Sc., (Agriculture) Mahathma Phule Agriculture University, 1998 __________________________________________________________ A THESIS Submitted in partial fulfillment of the requirements for the degree MASTER OF SCIENCE Department of Entomology College of Agriculture KANSAS STATE UNIVERSITY Manhattan, Kansas 2005 ABSTRACT The control of Indianmeal moth [Plodia interpunctella (Hübner)], a commonly found serious stored product pest around the world, relies mainly upon chemical control methods. Because of recent changes in the laws and regulations governing pesticide usage in the United States, there is an increasing need for finding safer chemicals to control insect pests. Hydroprene, an insect growth regulator, is considered to be a safe alternative. In this study, I quantified the effects of hydroprene on two critical life stages of Indianmeal moth, the eggs and 5th instar wandering phase larvae. Maximum development time in the untreated controls was 13.6 ± 0.6 d at 16°C and minimum development time was 2.3 ± 0.4 d at 32°C. At 20°C and 24°C, the effect of hydroprene on egg development became more evident; development time generally increased with exposure interval, with some variability in the data. The mean egg mortality among all temperatures was 7.3 ± 4.6%. Among the treatments, mortality of eggs increased as the exposure periods increased within any given temperature, with a dramatic increase in mortality with increase in temperature. Egg mortality was lowest at 16°C when exposed for 1 h (0 ± 3%), but mortality gradually increased up to 32 ± 3% when exposed for 18 h.
    [Show full text]
  • Invited Talk
    National Conference on Frontiers in Plant Physiology Towards Sustainable Agriculture, 5-7 November, 2009, Jorhat, Assam Invited Talk 1 National Conference on Frontiers in Plant Physiology Towards Sustainable Agriculture, 5-7 November, 2009, Jorhat, Assam SINGLE CROSS HYBRID TECHNOLOGY FOR MANAGING ABIOTIC STRESSES IN MAIZE Sain Dass and Ishwar Singh Directorate of Maize Research, Pusa Campus, New Delhi 110012 Maize (Zea mays L.) is the most widely cultivated crop of the world being grown in tropics, sub-tropics and temperate regions under irrigated to semi-arid conditions. It is crop of more than 150 counties. The major maize growing countries are USA, China, Brazil, Mexico, France and India. It is third most important cereal crop in India after rice and wheat that occupies about 8.12 million hectares having production of 19.3 million tones with average productivity of 2.37 tonnes/ ha during 2007-08. It is an important crop that holds a unique position in world agriculture, for food (25%), animal feed (12%), and poultry feed (49%), starch (12%), brewery (1%) and seed 1%. Apart from normal maize it has many other types viz. quality protein maize (QPM), sweet corn, baby corn, pop corn, waxy corn, high oil, etc. It generates employment for many million people in the world. Single cross hybrids (SCH) are most productive among the other types of hybrids and composite varieties of maize. Productive inbred lines are the strength of hybrid development programme. As many as 44 single cross hybrids of different maturity have been developed in a short span of period for different agro-ecological conditions.
    [Show full text]
  • Rashid Mumtaz
    BIO EFFICACY OF SOME PLANT EXTRACTS AGAINST SUGARCANE STALK BORER CHILD AURICILIUS DUDGEON (LEPIDOPTERA: CRAMBIDAE) ^ •••-.X V X ABSTRACT '\ THESIS ^^ SUBMITTED FOR THE AWARD OF THE DEGREE OF ZOOLOGY '^p-' (ENTOMOLOGY) .^ BY RASHID MUMTAZ DEPARTMENT OF ZDaLOCSY AL.IC3ARH MUSLIM UIMIVERSITY ALJGARH ((NiOfA) 2005 ABSTRACT Bio efficacy of some plant extracts against sugarcane stalk borer Chilo auricilius Dudgeon (Lepidoptera: Crambidae) Sugarcane is liable to attack by insect pests right from planting till harvest. The pests cause enormous losses both in tonnage and recovery of sugar in the mill. It is estimated about 8 to 10 percent of total sugarcane crop in the country is damaged by insects every year and this works out to a loss of 80 to 100 corers of rupees. The occurrence of drought and flood influenced the heavy buildup certain pests over large areas in epidemic from. (Siddiqui, 1960; Verma et, al. 1991). In general, the insect pest problems of sugarcane are more sever in subtropical belt than the fropical belt. Widespread use of synthetic pesticides has led to the development of pesticidal resistance in a large number of insect pests (Saxena and Khan, 1986). There has been an urgent need for selective and effective pest control methods which can be easily included in the modem pest management concept (Back, 1972) new bioactive products will be needed which are pest specific. Ecofriendly to man and other beneficial organisms biodegradable, less prone to development of pest resistance and economical viable. Insecticides of plant origin provided a substantial answer to many of the problems particularly the environmental pollution and ecofriendly in nature.
    [Show full text]
  • Combined Effect of Local Isolate Spodoptera Littoralis Nucleopolyhedrosis Virus and Bacillus Thuringiensis on Culex Pipiens L. L
    The Journal of Basic & Applied Zoology (2012) 65, 74–78 The Egyptian German Society for Zoology The Journal of Basic & Applied Zoology www.egsz.org www.sciencedirect.com Combined effect of local isolate Spodoptera littoralis nucleopolyhedrosis virus and Bacillus thuringiensis on Culex pipiens L. larvae (Culicidae: Diptera) Dalia M. Mahmoud 1, Marah M. Abd El-Bar *, Magda H. Abdul Aziz Radi 2 Department of Entomology, Faculty of Science, Ain Shams University, Abbassya, Cairo, Egypt Received 13 June 2012; accepted 25 August 2012 Available online 29 October 2012 KEYWORDS Abstract A local isolate of Spodoptera littoralis nucleopolyhedrosis virus (SlNPV) showed patho- Culex pipiens; genicity against Culex pipiens larvae, its long incubation period was broken using a combination Nucleopolyhedrosis virus; between SlNPV and Bacillus thuringiensis subsp. kurstaki (Btk). Laboratory bioassay tests revealed Bacillus thuringiensis; that Culex pipiens 3rd larval instars were susceptible to the applied combination. Data revealed that Combination; the addition of Btk generally increased the toxic effect of SlNPV since LC50 decreased from Biocontrol 1.3 · 103 PIB/ml (for SlNPV alone) to 3.6 · 102 mixed polyhedra and spores/ml (SlNPV-Btk com- bination), indicating a synergistic ratio of 3.6. LC95 was surprisingly 137-folds dropped as well. The addition of SlNPV generally increased the toxic effect of Btk at a low concentration, where the LC50 outstandingly decreased from 2.5 · 105 spores/ml (for Btk) to 3.6 · 102 mixed polyhedra and spores/ml (for SlNPV-Btk combination), indicating a synergistic ratio of 6.9 · 102. ª 2012 The Egyptian German Society for Zoology. Production and hosting by Elsevier B.V.
    [Show full text]
  • Isolation and Characterization of Baculoviruses from Major
    Journal of Entomology and Zoology Studies 2020; 8(5): 122-126 E-ISSN: 2320-7078 P-ISSN: 2349-6800 Isolation and characterization of baculoviruses www.entomoljournal.com JEZS 2020; 8(5): 122-126 from major lepidopteran insect pests infesting © 2020 JEZS Received: 16-07-2020 jute, Corchorus olitorius Linn Accepted: 18-08-2020 V Ramesh Babu V Ramesh Babu, G Sivakumar, S Satpathy and BS Gotyal Crop Protection Division, I.C.A.R- Central Research Institute for Jute and Allied Abstract Fibres, Barrackpore, Kolkata, Jute hairy caterpillar, Spilosoma obliqua Walker (Arctiidae: Lepidoptera) and jute semilooper, Anomis West Bengal, India sabulifera Guenee (Noctuidae: Lepidoptera) duo are the important lepidopteran insect pests infesting jute and causes damage by defoliation. Epizootic caused by a nucleopolyhedrovirus (NPV) was observed in G Sivakumar the jute field infested with S. obliqua and A. sabulifera during a routine survey in farmer’s fields. The Division of Germplasm, NPV was isolated and characterized based on polyhedrin gene of NPV which resulted in an amplicon Conservation and Utilisation, size of 700bp each for the both SpobNPV and AsNPV. Bioassay studies with second instar larvae of A. 4 I.C.A.R- National Bureau of sabulifera and S. obliqua revealed the median lethal concentration (LC50) of AsNPV as 5.37 × 10 Agriculturally Important OBs/ml and 2.44 x 104 OBs/ml at 72 HAT (Hours After Treatment). The SpobNPV at the highest POB Insects, Hebbal, Bengaluru, count @3.2 x106 OBs/ml caused cent per cent larval mortality, similarly AsNPV with POB (Poly Karnataka, India Occlusion Bodies) concentration @2.8 x106 OBs/ml resulted in 83 per cent larval mortality.
    [Show full text]
  • EU Project Number 613678
    EU project number 613678 Strategies to develop effective, innovative and practical approaches to protect major European fruit crops from pests and pathogens Work package 1. Pathways of introduction of fruit pests and pathogens Deliverable 1.3. PART 7 - REPORT on Oranges and Mandarins – Fruit pathway and Alert List Partners involved: EPPO (Grousset F, Petter F, Suffert M) and JKI (Steffen K, Wilstermann A, Schrader G). This document should be cited as ‘Grousset F, Wistermann A, Steffen K, Petter F, Schrader G, Suffert M (2016) DROPSA Deliverable 1.3 Report for Oranges and Mandarins – Fruit pathway and Alert List’. An Excel file containing supporting information is available at https://upload.eppo.int/download/112o3f5b0c014 DROPSA is funded by the European Union’s Seventh Framework Programme for research, technological development and demonstration (grant agreement no. 613678). www.dropsaproject.eu [email protected] DROPSA DELIVERABLE REPORT on ORANGES AND MANDARINS – Fruit pathway and Alert List 1. Introduction ............................................................................................................................................... 2 1.1 Background on oranges and mandarins ..................................................................................................... 2 1.2 Data on production and trade of orange and mandarin fruit ........................................................................ 5 1.3 Characteristics of the pathway ‘orange and mandarin fruit’ .......................................................................
    [Show full text]
  • Moths (Lepidoptera) Diversity of District JEZS 2018; 6(2): 1253-1263 © 2018 JEZS Koderma, Jharkhand Received: 05-01-2018 Accepted: 06-02-2018
    Journal of Entomology and Zoology Studies 2018; 6(2): 1253-1263 E-ISSN: 2320-7078 P-ISSN: 2349-6800 Moths (Lepidoptera) diversity of district JEZS 2018; 6(2): 1253-1263 © 2018 JEZS Koderma, Jharkhand Received: 05-01-2018 Accepted: 06-02-2018 Navneet Singh Navneet Singh, Jalil Ahmad and Rahul Joshi Zoological Survey of India, Kolkata, New Alipore, West Abstract Bengal, India The manuscript is about the moths collected from Koderma district of Jharkhand state. Koderma is Jalil Ahmad unexplored as far as its floral and faunal composition is concerned. The information is based on four Zoological Survey of India, surveys conducted in July 2012, August 2013, September 2014 and October 2015. A total of 140 species Gangetic Plains Regional Centre, under 106 genera of moths are reported from Koderma. Out of which, 32 species are reported for the first Sector-8, Bahadurpur Housing time from Jharkhand. Family Erebidae dominated among all the 17 families in diversity as well as in Colony, Patna, Bihar, India abundance. Rahul Joshi Keywords: lepidoptera, moths, diversity, new records, koderma Zoological Survey of India, Gangetic Plains Regional Centre, Introduction Sector-8, Bahadurpur Housing Colony, Patna, Bihar, India The study area Koderma was selected due to its unique geographical location. It is the northern edge of Chhota Nagpur plateau and from where the plateau smoothly disappears into the Gangetic plains of Bihar. The area is also known as a lower Hazaribagh plateau. Koderma lies between 24015.46 & 2404918 N latitude and 8502601 & 8505416 east longitude with an average altitude of 397m above the sea level. It covers an area of about 1500.00 sq kms [1].
    [Show full text]
  • Morphological Characterization and Genetic Divergence in Oleiferous Brassica Species
    FARMERS PRACTICES FOR COMBATING INSECT PESTS OF JUTE IN THREE INTENSIVE JUTE GROWING AREAS IN BANGLADESH AND THEIR ON-STATION VALIDATION TO DEVELOP AN IPM PACKAGE UTILIZING BIO-CONTROL AGENT AND NEEM PRODUCTS BY MD. AZIZUL HAQ DEPARTMENT OF ENTOMOLOGY SHER-E-BANGLA AGRICULTURAL UNIVERSITY DHAKA-1207 JUNE, 2011 FARMERS PRACTICES FOR COMBATING INSECT PESTS OF JUTE IN THREE INTENSIVE JUTE GROWING AREAS IN BANGLADESH AND THEIR ON-STATION VALIDATION TO DEVELOP AN IPM PACKAGE UTILIZING BIO-CONTROL AGENT AND NEEM PRODUCTS BY MD. AZIZUL HAQ REG. NO.: 05-01580 A thesis submitted to the Faculty of Agriculture, Sher-e-Bangla Agricultural University, Dhaka in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE IN ENTOMOLOGY SEMESTER: JANUARY-JUNE, 2011 Approved by: Prof. Dr. Md. Mizanur Rahman Ayesha Akter Supervisor Co-Supervisor & Assistant Professor Prof. Dr. Md. Razzab Ali Chairman Examination Committee DEDICATED TO MY BELOVED PARENTS DEPARTMENT OF ENTOMOLOGY Sher-e-Bangla Agricultural University Sher-e-Bangla Nagar, Dhaka-1207 Memo No: SAU/Entomology/ CERTIFICATE This is to certify that thesis entitled, “Farmers practices for combating insect pests of jute in three intensive jute growing areas in Bangladesh and their on-station validation to develop an IPM package utilizing bio-control agent and neem products” submitted to the Faculty of Agriculture, Sher-e-Bangla Agricultural University, Dhaka, in partial fulfillment of the requirements for the degree of Master of Science in Entomology, embodies the result of a piece of bona fide research work carried out by Md. Azizul Haq, Registration No. 05-01580 under my supervision and guidance.
    [Show full text]