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Transformation of azuki bean by Agrobacterium tumefaciens

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Abstract

Stable transformation and regeneration was developed for a grain legume, azuki bean (Vigna angularis Willd. Ohwi & Ohashi). Two constructs containing the neomycin phosphotransferase II gene (nptII) and either the β-glucuronidase (GUS) gene or the modified green fluorescent protein [sGFP(S65T)] gene were introduced independently via Agrobacterium tumefaciens-mediated transformation. After 2 days of co-cultivation on MS medium supplemented with 100 μM acetosyringone and 10 mg l−1 6-benzyladenine, seedling epicotyl explants were placed on regeneration medium containing 100 mg l−1 kanamycin. Adventitious shoots developing from explant calli were excised onto rooting medium containing 100 mg l−1 kanamycin. Rooted shoots were excised and repeatedly selected on the same medium containing kanamycin. Surviving plants were transferred to soil and grown in a green house to produce viable seeds. This process took 5 to 7 months after co-cultivation. Molecular analysis confirmed the stable integration and expression of foreign genes.

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Authors and Affiliations

  1. School of Agricultural Sciences, Nagoya University, Chikusa, Nagoya, 464-01, Japan

    Tsuyoshi Yamada & Kazumi Hattori

  2. National Agriculture Research Center, Tsukuba, Ibaraki, 305-8666, Japan

    Masayoshi Teraishi

  3. Chugoku National Agricultural Experiment Station, Fukuyama, Hiroshima, 721-8514, Japan

    Masao Ishimoto

Authors
  1. Tsuyoshi Yamada
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  2. Masayoshi Teraishi
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  3. Kazumi Hattori
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  4. Masao Ishimoto
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Correspondence to Masao Ishimoto.

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Yamada, T., Teraishi, M., Hattori, K. et al. Transformation of azuki bean by Agrobacterium tumefaciens. Plant Cell, Tissue and Organ Culture 64, 47–54 (2001). https://doi.org/10.1023/A:1010635832468

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  • DOI: https://doi.org/10.1023/A:1010635832468

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