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Screening, Diversity and Partial Sequence Comparison of Vegetative Insecticidal Protein (vip3A) Genes in the Local Isolates of Bacillus thuringiensis Berliner

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Abstract

Characterization, direct sequencing of the PCR amplicon and phylogenetic relationship was done to discover a novel Vip protein genes of the Bt isolates, to improve the prospects for insect control, more Vip proteins should be sought out and researched to predict their insecticidal activity. Characterization was based on direct sequencing of PCR amplicon using primers specific to vip3A gene was presented here. 12 out of 18 isolates screened were positive for vip gene-specific primers. Homology search for the partial sequences using BLAST showed that 11 isolates had high similarity to vip3Aa gene and only one fragment with vip3Ae gene (25–100% at nucleotide and amino acid level). Phylogenetic analysis showed that the gene sequences were responsible for geographic separation for divergence within vip genes, consistent with the evaluation of distinct bacterial population. Despite the geographical distances, strains harbouring vip genes have originated from common ancestors may significantly contribute to control resistant insect pests. Some strains have evolved to be quite distinct and others remain as members of closely related groups. The reported method is a powerful tool to find novel Vip3A proteins from large-scale Bt strains which is effective in terms of time and cost. Further the Vip proteins produced by different strains of B. thuringiensis are unique in terms of the sequence divergence and hence may also differ in their insecticidal activities.

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Acknowledgments

Infrastructure facility and encouragement by The Director, Indian Institute of Horticultural Research (IIHR) is duly acknowledged. The authors are grateful to ICAR, New Delhi for funding this study under Network project on Application of Microbes in Agriculture and Allied Sectors (AMAAS).

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Authors and Affiliations

  1. Bio Pesticide Laboratory (BPL), Division of Biotechnology, Indian Institute of Horticultural Research (IIHR), Hessarghatta Lake Post, Bangalore, 560 089, Karnataka, India

    R. Asokan & H. M. Mahadeva Swamy

  2. National Bureau of Agriculturally Important Micro Organisms (NBAIM), Mau Nath Bhanjan, 275101, Uttar Pradesh, India

    D. K. Arora

Authors
  1. R. Asokan
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  2. H. M. Mahadeva Swamy
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  3. D. K. Arora
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Correspondence to R. Asokan or H. M. Mahadeva Swamy.

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Asokan, R., Swamy, H.M.M. & Arora, D.K. Screening, Diversity and Partial Sequence Comparison of Vegetative Insecticidal Protein (vip3A) Genes in the Local Isolates of Bacillus thuringiensis Berliner. Curr Microbiol 64, 365–370 (2012). https://doi.org/10.1007/s00284-011-0078-z

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