Transformation of azuki bean by Agrobacterium tumefaciens

  • Yamada T
  • Teraishi M
  • Hattori K
 et al. 
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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 beta - 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 muM acetosyringone and 10 mg 1(-1) 6-benzyladenine, seedling epicotyl explants were placed on regeneration medium containing 100 mg 1(-1) kanamycin. Adventitious shoots developing from explant calli were excised onto rooting medium containing 100 mg 1(-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.

Author-supplied keywords

  • 6-benzyladenine
  • Acetosyringone
  • Co-cultivation
  • Grain legume
  • Green fluorescent protein
  • Neomycin phosphotransferase
  • Transgenic plant
  • Vigna angularis

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