Distribution of vip Genes, Protein Profiling and Determination of Entomopathogenic Potential of Local Isolates of Bacillus thuringiensis

The aim of this study was to screen the Bacillus thuringiensis ( Bt ) strains isolated from local ecological niche for the presence of vegetative insecticidal protein ( vip ) genes and its characterization. PCR screening was carried out to determine different vip genes viz., vip1 / vip2 and vip3 genes, harboured by native Bt isolates using four gene specific primers. The PCR results depicted the presence of all three vip -type genes in these isolates. Strains containing vip3 -type genes were the most abundant (12.5%) in local isolates, followed by vip1 / vip2 -type genes (10%). Insect bioassays for these proteins were conducted against Helicoverpa armigera ( H. armigera ). Four Bt isolates PDKV-08 (161.80), PDKV-21 (173.62), NCIM-5110 (170.57) and NCIM-5132 (178.61) depict lower LC 50 values against H. armigera than the standard HD-1 (190.42). The present foundation work can be used for the production of more effective Bt based biopesticide and the identified genes can be explored for the production of next generation insect resistant transgenic plants.

[1]  P. Kaur,et al.  Inheritance of resistance to Bacillus thuringiensis Cry1Ac toxin in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) from India. , 2011, Pest management science.

[2]  G. Gujar,et al.  Field-evolved resistance to Bt toxin Cry1Ac in the pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), from India. , 2011, Pest management science.

[3]  B. Tabashnik,et al.  Field-Evolved Resistance to Bt Cotton: Bollworm in the U.S. and Pink Bollworm in India , 2010 .

[4]  M. Soberón,et al.  How to cope with insect resistance to Bt toxins? , 2008, Trends in biotechnology.

[5]  R. Akhurst,et al.  Unusually High Frequency of Genes Encoding Vegetative Insecticidal Proteins in an Australian Bacillus thuringiensis Collection , 2008, Current Microbiology.

[6]  Juhua Chang,et al.  Resistance Allele Frequency to Bt Cotton in Field Populations of Helicoverpa armigera (Lepidoptera: Noctuidae) in China , 2008, Journal of economic entomology.

[7]  D. Crowder,et al.  Insect resistance to Bt crops: evidence versus theory , 2008, Nature Biotechnology.

[8]  S. Downes,et al.  Frequency of Alleles Conferring Resistance to the Bt Toxins Cry1Ac and Cry2Ab in Australian Populations of Helicoverpa armigera (Lepidoptera: Noctuidae) , 2007, Journal of economic entomology.

[9]  Qiyu Bao,et al.  Evidence for positive Darwinian selection of Vip gene in Bacillus thuringiensis. , 2007, Journal of genetics and genomics = Yi chuan xue bao.

[10]  C. Berry,et al.  Screening of Brazilian Bacillus thuringiensis isolates active against Spodoptera frugiperda, Plutella xylostella and Anticarsia gemmatalis , 2007 .

[11]  J. B.J. van Rensburg,et al.  First report of field resistance by the stem borer, Busseola fusca (Fuller) to Bt-transgenic maize , 2007 .

[12]  M. Yuan,et al.  `Cloning of vip1/vip2 genes and expression of Vip1Ca/Vip2Ac proteins in Bacillus thuringiensis , 2007 .

[13]  A. Mandaokar,et al.  Isolation, characterization and expression of a novel vegetative insecticidal protein gene of Bacillus thuringiensis. , 2005, FEMS microbiology letters.

[14]  X. Guan,et al.  Cloning and localization of vip3A gene of Bacillus thuringiensis , 2004, Biotechnology Letters.

[15]  L. Loguercio,et al.  Combined analysis of supernatant‐based feeding bioassays and PCR as a first‐tier screening strategy for Vip ‐derived activities in Bacillus thuringiensis strains effective against tropical fall armyworm , 2002, Journal of applied microbiology.

[16]  S. K. Jalali,et al.  Toxicity Analysis of N- and C-Terminus-Deleted Vegetative Insecticidal Protein from Bacillus thuringiensis , 2001, Applied and Environmental Microbiology.

[17]  K. Wilson Preparation of Genomic DNA from Bacteria , 2001, Current protocols in molecular biology.

[18]  J. Tainer,et al.  Evolution and mechanism from structures of an ADP-ribosylating toxin and NAD complex , 1999, Nature Structural Biology.

[19]  W. C. Rice Specific primers for the detection of vip3A insecticidal gene within a Bacillus thuringiensis collection , 1999 .

[20]  G. W. Warren Vegetative Insecticidal Proteins: Novel Proteins for Control of Corn Pests , 1997 .

[21]  M. Koziel,et al.  Vip3A, a novel Bacillus thuringiensis vegetative insecticidal protein with a wide spectrum of activities against lepidopteran insects. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[22]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[23]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[24]  J. Lindsey Flexner,et al.  on Microbial Insecticides , 2022 .