Transformation of azuki bean by Agrobacterium tumefaciens

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 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.