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Variant cry1Ab entomocidal Bacillus thuringiensis toxin gene facilitates the recovery of an increased number of lepidopteran insect resistant independent rice transformants against yellow stem borer (Scirpophaga incertulus) inflicted damage

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Abstract

The primary technical constraint plant scientists face in generating insect resistant transgenic crops with insecticidal Bacillus thuringiensis (Bt) crystal protein (Cry) genes remains failing to generate sufficiently large numbers of effective resistant transgenic plant lines. One possible means to overcome this challenge is through deployment of a Cry toxin gene that contains high levels of insecticidal specific activity for target insect pests. In the present study, we tested this hypothesis using a natural variant of the Cry1Ab toxin under laboratory conditions that possessed increased insecticidal potency against the yellow stem borer (YSB, Scirpophaga incertulus), one of the most damaging rice insect pests. Following adoption of a stringent selection strategy for YSB resistant transgenic rice lines under field conditions, results showed recovery of a significantly higher number of YSB resistant independent transgenic plant lines with the variant cry1Ab gene relative to transgenic plant lines harbouring cry1Ab berliner gene. Structural homology modelling of the variant toxin peptide with the Cry1Aa toxin molecule, circular dichroism spectral analysis, and hydropathy plot analysis indicated that serine substitution by phenylalanine at amino acid position 223 of the Cry1Ab toxin molecule resulted in a changed role for α-helix 7 in domain I of Cry1Ab for enhanced toxicity.

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Abbreviations

EDTA:

Ethylene diamine tetra-acetate

SDS:

Sodium dodecyl sulphate

SDS-PAGE:

Sodium dodecyl sulphate polyacrylamide gel electrophoresis

Tris:

Tris (hydroxymethyl) amino methane

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Acknowledgments

Authors appreciate the technical support received from Messrs Bijon Ghosh for skilful insect handling; Partha Das and Sona Dogra for their expertise in rice transformation; Sudarshan Maity and Manoj Aditya for taking care of experimental materials in containment condition; and finally Meghnath Prasad for his secretarial support.

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Author notes

  1. Abhijit Dandapat

    Present address: University of Minnesota, 312 Church St SE, NHH, Room 4062, Minneapolis, MN, 55455, USA

  2. Anannya Banga

    Present address: Stem Cell Institute, University of Minnesota, Minneapolis, 55455, USA

  3. Chandi Charan Mandal

    Present address: Department of Pathology, University of Texas Health Science Centre at San Antonio, San Antonio, TX, 78 229, USA

  4. Munshi Azad Hossain

    Present address: Division of Biological Sciences, University of California, San Diego, La Jolla, CA, 92093, USA

Authors and Affiliations

  1. Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology, Kharagpur, 721302, India

    Abhijit Dandapat, Jagannath Bhattacharyya, Srimonta Gayen, Anirban Chakraborty, Anannya Banga, Rajeswari Mukherjee, Chandi Charan Mandal, Munshi Azad Hossain, Samarjit Roy, Asitava Basu & Soumitra Kumar Sen

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  1. Abhijit Dandapat
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  2. Jagannath Bhattacharyya
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  10. Asitava Basu
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Corresponding author

Correspondence to Soumitra Kumar Sen.

Additional information

Abhijit Dandapat and Jagannath Bhattacharyya contributed equally.

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Dandapat, A., Bhattacharyya, J., Gayen, S. et al. Variant cry1Ab entomocidal Bacillus thuringiensis toxin gene facilitates the recovery of an increased number of lepidopteran insect resistant independent rice transformants against yellow stem borer (Scirpophaga incertulus) inflicted damage. J. Plant Biochem. Biotechnol. 23, 81–92 (2014). https://doi.org/10.1007/s13562-013-0190-1

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