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Stable genetic transformation of Vigna mungo L. Hepper via Agrobacterium tumefaciens

  • Genetic Transformation and Hybridization
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An Erratum to this article was published on 04 July 2003

Abstract

Vigna mungo is one of the large-seeded grain legumes that has not yet been transformed. We report here for the first time the production of morphologically normal and fertile transgenic plants from cotyledonary-node explants inoculated with Agrobacterium tumefaciens carrying binary vector pCAMBIA2301, the latter of which contains a neomycin phosphotransferase (nptII) gene and a β-glucuronidase (GUS) gene (uidA) interrupted with an intron. The transformed green shoots, selected and rooted on medium containing kanamycin, tested positive for nptII and uidA genes by polymerase chain reaction (PCR) analysis. These shoots were established in soil and grown to maturity to collect the seeds. Mechanical wounding of the explants prior to inoculation with Agrobacterium, time lag in regeneration due to removal of the cotyledons from explants and a second round of selection at the rooting stage were found to be critical for transformation. Analysis of T0 plants showed the expression and integration of uidA into the plant genome. GUS activity in leaves, roots, flowers, anthers and pollen grains was detected by histochemical assay. PCR analysis of T1 progeny revealed a Mendelian transgene inheritance pattern. The transformation frequency was 1%, and 6–8 weeks were required for the generation of transgenics.

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Fig. 1.
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Abbreviations

BA :

6-Benzylaminopurine

IBA :

Indole-3-butyric acid

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Acknowledgements

Our research programme is generously supported by the Department of Biotechnology, New Delhi. We are grateful to the Center for Application of Molecular Biology to International Agriculture (CAMBIA), Australia for plasmid CAMBIA2301 and to Prof. Anil Grover, New Delhi for providing laboratory facilities for the Southern analysis. Raman Saini and Sonia Jaiwal are grateful to DBT and CSIR, New Delhi, respectively, for the award of a Senior Research Fellowship.

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Authors and Affiliations

  1. Department of Biosciences, M.D. University, 124001, Rohtak, India

    R. Saini, S. Jaiwal & P. K. Jaiwal

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  1. R. Saini
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  2. S. Jaiwal
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  3. P. K. Jaiwal
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Correspondence to P. K. Jaiwal.

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Communicated by G.C. Phillips

An erratum to this article is available at http://dx.doi.org/10.1007/s00299-003-0643-4.

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Saini, R., Jaiwal, S. & Jaiwal, P.K. Stable genetic transformation of Vigna mungo L. Hepper via Agrobacterium tumefaciens . Plant Cell Rep 21, 851–859 (2003). https://doi.org/10.1007/s00299-003-0574-0

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