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Formulation and optimisation of various nuclear polyhedrosis virus isolates and assessment of their insecticidal activity against Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) larvae
Authors:Elangovan Elamathi  J. RajaRajaCholan  N. Vijayakumar  A. Ramamourti
Affiliation:1. Department of Microbiology , Tamil Nadu Agricultural University , Coimbatore , 641003 , India egelamathy@gmail.com;3. Centre for Plant Breeding and Genetics , Tamil Nadu Agricultural University , Coimbatore , 641003 , India;4. Perunthalaivar Kamaraj Krish Vigyan Kendra (PKKVK) , Kurumbapet , Puducherry , 605009 , India
Abstract:Helicoverpa armigera (Hubner) is a pest that poses serious threat to the tomato crop in India. Larvae of this species are susceptible to the nucleopolyhedrovirus (NPV), which has attracted interest as a potential biocontrol agent. Rearing of larvae in natural diet resulted in less number of pupae, while in artificial diet more pupae and healthy adults were obtained. The NPV was tested for its insecticidal action against H. armigera in natural and artificial diets. The bioassay results of inoculated H. armigera nucleopolyhedrosis virus (HaNPV) with different concentrations indicate that the 4.0 g/l dosage caused maximum mortality (70.3% and 60.54%), and minimum mortality 46.83% and 44.08% was recorded in the 0.5 g/l dosage under laboratory and pot culture conditions, respectively. Polyhedral inclusion bodies (PIBs) were estimated using a standard haemocytometer. According to the estimate, purified PIBs were found at a concentration of 1 × 109/ml. The different formulation of NPV, i.e. wettable fine powder with a hue of white, had a particle size of 100 μm with a concentration of 1 × 108/ml. The encapsulated product was found to be in the form of beads and was red in colour with a concentration of 1 × 108 PIBs/ml. The effectiveness of NPV against H. armigera larvae was noted at maximum LC50 values at a concentration of 1 × 109/ml. Qualitative analysis estimates showed that protein polyhedrin and enzyme chitinase had a molecular weight of 33 and 45 kDa, respectively. Viral DNA was isolated from NPV-infected larvae and was then separated using SDS-PAGE. The estimated polyhedrin with a molecular weight of 33 kDa, which is responsible for the increased mortality percent of larvae, was confirmed by the bioassay. Hence it was concluded that NPV is ecofriendly in the management of pests on tomato and other crops.
Keywords:Helicoverpa armigera  nucleopolyhedrovirus  HaNPV  media optimisation  formulation  occlusion bodies  virus infectivity  bioassay  qualitative analysis
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