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Over-expression of Trigonella foenum-graecum defensin (Tfgd2) and Raphanus sativus antifungal protein (RsAFP2) in transgenic pigeonpea confers resistance to the Helicoverpa armigera

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Abstract

Pigeonpea is an important food legume crop cultivated in tropical and sub-tropical regions around the world wherein the Indian subcontinent accounting for over 90 % of global production. It is a rich source of protein and is an important component of a well-balanced diet for the majority of Indians. Among the other insect pests, pigeonpea productivity is mostly affected by Helicoverpa armigera which is causing severe yield loss. Non-availability of resistant genes in germplasm and constraints with traditional breeding induce the application of a genetic engineering approach to generate insect resistance in pigeonpea. Expression of plant defensins in various crops provided enhanced resistance towards a variety of pests and pathogens. In the current study, two defensins Trigonella foenum-graecum defensin 2 (Tfgd2) and Raphanus sativus antifungal protein 2 (RsAFP2) integrated by a linker peptide was transferred into pigeonpea as a fusion gene by Agrobacterium mediated transformation. Putative transgenic lines were confirmed through PCR and the promising lines were identified in the following generations based upon integration, expression and bioefficiency of the fusion gene. Leaf bioassay conducted against H. armigera larvae showed increased levels of insect resistance compared to the control, where six T2 plants were identified as superior lines showing less than 25 % of leaf damage. Our findings illustrates that Tfgd2–RsAFP2 fusion protein is efficient in imparting protection against the insect pest and the transgenic lines developed in this study could be used for further pigeonpea improvement projects.

Key message

Over-expression of Tfgd2-RsAFP2 fusion gene conferred enhanced insect resistance against Helicoverpa armigera in transgenic pigeonpea plants.

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Data availability

The authors confirm that the data supporting the findings of this study are available within the article and its supplementary material. Data derived supporting the findings of this study are available from the corresponding author on request.

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Acknowledgements

The authors are grateful to Science & Engineering Research Board (SERB)–Young Scientist Scheme (YSS) (SB/YS/LS-83/2014 to Dr. K. Vasavi Rama) for their encouragement and financial support to this project. The authors are thankful to Prof. P. B. Kirti, Department of Plant Sciences, University of Hyderabad for his continuous guidance and valuable support to complete this work. The authors also acknowledge the support of Department of Biotechnology, GITAM School of Technology and Department of Biotechnology, GITAM School of Science, GITAM (Deemed to be University), Visakhapatnam in successful completion of this study.

Funding

This study was financially endorsed by grants from Science and Engineering Research Board (SERB), Department of Science & Technology, Government of India (SERB-YSS (SB/YS/LS-83/2014 to Dr. K. Vasavi Rama).

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  1. Department of Biotechnology, GITAM School of Technology, GITAM (Deemed to be University), Visakhapatnam, Andhra Pradesh, 530045, India

    Vasavirama Karri

  2. Department of Biotechnology, GITAM School of Science, GITAM (Deemed to be University), Visakhapatnam, Andhra Pradesh, 530045, India

    Nirmala Nalluri

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VK designed and guided NN to work on this specific topic to acquire data. NN prepared manuscript according to guidelines under the supervision of VK. Finally the manuscript was checked and corrected by VK for submission in favor of publication. Both worked hard in the analysis of data to complete this investigation. All authors have read and approved the manuscript.

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Nalluri, N., Karri, V. Over-expression of Trigonella foenum-graecum defensin (Tfgd2) and Raphanus sativus antifungal protein (RsAFP2) in transgenic pigeonpea confers resistance to the Helicoverpa armigera. Plant Cell Tiss Organ Cult 152, 569–582 (2023). https://doi.org/10.1007/s11240-022-02431-0

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