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A kidney bean trypsin inhibitor with an insecticidal potential against Helicoverpa armigera and Spodoptera litura

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Abstract

In the present study, trypsin inhibitor extracts of ten kidney bean seed (Phaseolus vulgaris) varieties exhibiting trypsin and gut trypsin-like protease inhibitor activity were tested on Helicoverpa armigera and Spodoptera litura. Trypsin inhibitor protein was isolated and purified using multi-step strategy with a recovery of ~15 % and purification fold by ~39.4. SDS-PAGE revealed a single band corresponding to molecular mass of ~15 kDa and inhibitory activity was confirmed by reverse zymogram analyses. The inhibitor retained its inhibitory activity over a broad range of pH (3–11), temperature (40–60 °C) and thermostability was promoted by casein, CaCl2, BSA and sucrose. The purified inhibitor inhibited bovine trypsin in 1:1 molar ratio. Kinetic studies showed that the protein is a competitive inhibitor with an equilibrium dissociation constant of 1.85 μM. The purified trypsin inhibitor protein was further incorporated in the artificial diet and fed to second instar larvae. A maximum of 91.7 % inhibition was obtained in H. armigera, while it was moderate in S. litura (29 %) with slight varietal differences. The insect bioassay showed 40 and 22 % decrease in larval growth followed by 3 and 2 days delay in pupation of H. armigera and S. litura, respectively. Some of the adults emerged were deformed and not fully formed. Trypsin inhibitor protein was more effective against H. armigera as it showed 46.7 % mortality during larval growth period compared to S. litura (13.3 %).

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Acknowledgments

Ms. Anuradha Mittal greatly acknowledges the financial assistance from Department of Science and Technology, India as INSPIRE Junior Research Fellowship under INSPIRE Fellowship Scheme.

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Authors and Affiliations

  1. Department of Biochemistry, Kurukshetra University, Kurukshetra, 136119, India

    Anuradha Mittal & Vijay Kumar Gupta

  2. NRC on Plant Biotechnology, Lal Bahadur Shastri Building, PUSA Campus, New Delhi, India

    Rekha Kansal

  3. Division of Entomology, Indian Agricultural Research Institute, New Delhi, India

    Vinay Kalia & Monika Tripathi

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  1. Anuradha Mittal
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  2. Rekha Kansal
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  3. Vinay Kalia
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  4. Monika Tripathi
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  5. Vijay Kumar Gupta
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Correspondence to Vijay Kumar Gupta.

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Communicated by B. Barna.

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Mittal, A., Kansal, R., Kalia, V. et al. A kidney bean trypsin inhibitor with an insecticidal potential against Helicoverpa armigera and Spodoptera litura . Acta Physiol Plant 36, 525–539 (2014). https://doi.org/10.1007/s11738-013-1433-4

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  • DOI: https://doi.org/10.1007/s11738-013-1433-4

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