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Transgenic cowpea (Vigna unguiculata) seeds expressing a bean α-amylase inhibitor 1 confer resistance to storage pests, bruchid beetles

  • Genetic Transformation and Hybridization
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Abstract

Cowpea is one of the important grain legumes. Storage pests, Callosobruchus maculatus and C. chinensis cause severe damage to the cowpea seeds during storage. We employ a highly efficient Agrobacterium-mediated cowpea transformation method for introduction of the bean (Phaseolus vulgaris) α-amylase inhibitor-1 (αAI-1) gene into a commercially important Indian cowpea cultivar, Pusa Komal and generated fertile transgenic plants. The use of constitutive expression of additional vir genes in resident pSB1 vector in Agrobacterium strain LBA4404, thiol compounds during cocultivation and a geneticin based selection system resulted in twofold increase in stable transformation frequency. Expression of αAI-1 gene under bean phytohemagglutinin promoter results in accumulation of αAI-1 in transgenic seeds. The transgenic protein was active as an inhibitor of porcine α-amylase in vitro. Transgenic cowpeas expressing αAI-1 strongly inhibited the development of C. maculatus and C. chinensis in insect bioassays.

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Acknowledgments

We thank Commonwealth Scientific and Industrial Research Organization (CSIRO), Australia for providing the pTA3 (αAI-1 construct), Dr. T. J. V. Higgins, CSIRO Plant Industry, Australia, for his valuable suggestions and Prof. P. B. Kirti, University of Hyderabad, India for his generous technical assistance. We thank Dr. Toshihiko Komari, Japan Tobacco, Inc., Japan, for providing pSB1, Prof. K. Veluthambi, MKU, Madurai, India for the Agrobacterium strains and to the Center for Application of Molecular Biology to International Agriculture (CAMBIA), Australia for plasmid pCAMBIA2301. This work was funded by the Department of Biotechnology, Government of India. SKS and SB are grateful to Indian Institute of Technology Guwahati for a doctoral fellowship.

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Authors and Affiliations

  1. Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India

    Siva Kumar Solleti, Souvika Bakshi & Lingaraj Sahoo

  2. Center for Energy, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India

    Jubilee Purkayastha & Lingaraj Sahoo

  3. Department of Life Science, School of Life Sciences, Assam University, Silchar, 788 011, India

    Sanjib Kumar Panda

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  1. Siva Kumar Solleti
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  2. Souvika Bakshi
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  3. Jubilee Purkayastha
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  4. Sanjib Kumar Panda
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  5. Lingaraj Sahoo
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Correspondence to Lingaraj Sahoo.

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Communicated by R. Rose.

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Solleti, S.K., Bakshi, S., Purkayastha, J. et al. Transgenic cowpea (Vigna unguiculata) seeds expressing a bean α-amylase inhibitor 1 confer resistance to storage pests, bruchid beetles. Plant Cell Rep 27, 1841–1850 (2008). https://doi.org/10.1007/s00299-008-0606-x

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  • DOI: https://doi.org/10.1007/s00299-008-0606-x

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