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

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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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Authors and Affiliations

  1. Department of Plant Sciences, University of Hyderabad, Hyderabad, 500046, Telangana, India

    Akanksha Sharma, Vijayan Sambasivam, Pawan Shukla, Sakshi Rampuria & Pulugurtha Bharadwaja Kirti

  2. Tagore Government Arts and Science College, Puducherry, 605008, India

    Vijayan Sambasivam

  3. Central Sericultural Research and Training Institute, Central Silk Board, NH-1A, Gallandar, Pampore, J & K, 192 121, India

    Pawan Shukla

Authors
  1. Akanksha Sharma
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  2. Vijayan Sambasivam
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  3. Pawan Shukla
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  4. Sakshi Rampuria
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  5. Pulugurtha Bharadwaja Kirti
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Contributions

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 102, 1133–1143 (2020). https://doi.org/10.1007/s42161-020-00591-6

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