Characterization and Screening of Bacillus thuringiensis Isolated from Nashik and Ahmednagar Region of Maharashtra
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- Bacillus thuringiensis, Colony Morphology, Crystal Morphology, DNA Isolation, SDS-PAGE, Protein Harvesting
- Mane, V. A; Shinde, H. S; Pagar, T. A; Garud, M. P.
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- Bacillus thuringiensis is a Gram positive, facultative anaerobe, Soil dwelling, spore forming bacteria that produces toxic against different insect species. This feature makes it the widely used biological control agent in agriculture. The aim of this study was to isolate and cloned B. thuringiensis cry genes and check the toxicity level against Helicoverpa armigera. Soil samples were collected Nashik and Ahmednagar region of Maharashtra. The fried egg type colonies which were obtained from this soil sample were identified as Bacillus thuringiensis. The crystal morphology of isolates was observed under phase contrast microscope during spore formation and rectangular type crystal morphology was present in both the cultures. The bioassay was done to check mortality percentage in Helicoverpa armigera. For genotypic characterization, the cry gene content of the isolates was screened by polymerase chain reaction (PCR) analysis using universal primers as per Ben-Dov, et al., 1997. Both isolates were screened for cry genes and it was found that both isolates showing the band size more than the expected amplicon size, thus it was hypothesized that particular gene been amplified in different manner. So it is concluded that it may be a cry gene or Vegetative insecticidal Proteins (Vips) genes.
Full text: IJRAS_283_Final.pdf