An in vitro generated variant of Tephrosia villosa defensin (α-TvD1) enhances biotic stress tolerance in transgenic tobacco

Abstract

Plant defensins are small, cationic, cysteine-rich peptides with well conserved four disulphide bridges between an α-helix and three anti-parallel β-strands, produced by plants as a first line of innate immunity against the invading fungal pathogens. Some of the plant defensins exhibit strong antibacterial and anti-insect activities in vitro as well as in vivo. TvD1, a defensin from Tephrosia villosa was characterized in our previous studies with demonstrated inhibitory effect against many plant pathogenic fungi and some insects. To further enhance its activity, α-TvD1, a peptide variant of TvD1, was generated through site directed mutagenesis. It showed enhanced antifungal and Tenebrio molitor α-amylase inhibitory activity under in vitro condition. In the present study, we compared the antifungal and anti-insect activities of α-TvD1 with the native TvD1 in transgenic tobacco plants. α-TvD1 transgenic plants showed enhanced tolerance against the oomycetes pathogen Phytophthora parasitica var. nicotianae and the fungal pathogens Alternaria alternata and Rhizoctonia solani. Also, it showed enhanced anti-insect activity against the 1st and 2nd instar larvae of the generalist herbivore, Spodoptera litura. These results demonstrated that α-TvD1 appears to be a potent antimicrobial gene that can be used for generating superior transgenic crops with enhanced resistance against many phytopathogens and insect pests.

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Acknowledgements

The authors are grateful to AP-Netherlands Biotechnology Programme (rechristened as Agri Biotech Foundation, PJTS Agricultural University Campus, Hyderabad). The authors are grateful to the Facilities under DST-FIST, DBT-CREBB and UGC-SAP of the School of Life Sciences used in the Investigation. AS, VS, PS and SR acknowledge DBT, UGC, CSIR, and DST-INSPIRE, India for fellowships respectively.

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AS, VS, PBK conceived and designed the experiments. AS, PS performed the experiments. AS analyzed the data. SR contributed suggestions/reagents/materials/analysis tools. AS and VS wrote the manuscript. All the authors read and approved the manuscript.

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Correspondence to Vijayan Sambasivam or Pulugurtha Bharadwaja Kirti.

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Key message

Ectopic expression of α-TvD1, a peptide variant of Tephrosia villosa defensin imparts enhanced fungal tolerance in tobacco plants as compared to native TvD1 establishing its potential role in crop transformations.

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Sharma, A., Sambasivam, V., Shukla, P. et al. An in vitro generated variant of Tephrosia villosa defensin (α-TvD1) enhances biotic stress tolerance in transgenic tobacco. J Plant Pathol (2020). https://doi.org/10.1007/s42161-020-00591-6

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Keywords

  • Defensin variant
  • Transgenic tobacco plant
  • Native TvD1
  • Insect α-amylase
  • Necrotic lesion