Full Length Research Paper - Recovery of herbicide-resistant Azuki bean [Vigna angularis (Wild.), Ohwi & Ohashi] plants via Agrobacterium- mediated transformation
Transgenic azuki bean [Vigna angularis (Willd.) Ohwi &
Ohashi] plants expressing the hygromycin phosphotransferase (hpt),
green fluorescent protein (sgfp) and phosphinothricin acetyltransferase
(bar) genes were obtained by Agrobacterium - tumefacients -
mediated transformation. A total of 210 epicotyl explants were
inoculated with A. tumefaciens strain EHA105, harboring the binary
plasmid pZHBG on MS co-cultivation medium supplemented with 100 mM
acetosyringone and 10 mg/l of BA. Following selection on MS medium with
15 mg/l of hygromycin, the regenerated adventitious shoots that formed
on the induced calli were further screened for sgfp expression before
transferred to rooting medium. 31 transgenic plants were obtained with
transformation frequency of 14%. The presence of transgenes in
transformed azuki bean plants was confirmed by polymerase chain
reaction (PCR) and southern blot analysis. Transcription of the bar and
hpt genes was assessed by reverse transcription polymerase chain
reaction (RT-PCR) analysis. sgfp- positive transgenic plants exhibited
functional expression of the bar gene as determined by assaying for
resistance to bialaphos applied directly to leaves. This result
demonstrates the feasibility of introducing potentially useful
agronomic traits into azuki bean through genetic engineering