Journal
PLANT CELL REPORTS
Volume 26, Issue 2, Pages 187-198Publisher
SPRINGER
DOI: 10.1007/s00299-006-0224-4
Keywords
Agrobacterium tumefaciens; alpha-amylase inhibitor; genetic transformation; herbicide; mungbean; phosphinothricin; Vigna radiata
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Morphologically normal and fertile transgenic plants of mungbean with two transgenes, bar and alpha-amylase inhibitor, have been developed for the first time. Cotyledonary node explants were transformed by cocultivation with Agrobacterium tumefaciens strain EHA105 harboring a binary vector pKSB that carried bialaphos resistance (bar) gene and Phaseolus vulgaris alpha-amylase inhibitor-1 (alpha AI-1) gene. Green transformed shoots were regenerated and rooted on medium containing phosphinothricin (PPT). Preculture and wounding of the explants, presence of acetosyringone and PPT-based selection of transformants played significant role in enhancing transformation frequency. Presence and expression of the bar gene in primary transformants was evidenced by PCR-Southern analysis and PPT leaf paint assay, respectively. Integration of the Phaseolus vulgaris alpha-amylase inhibitor gene was confirmed by Southern blot analysis. PCR analysis revealed inheritance of both the transgenes in most of the T-1 lines. Tolerance to herbicide was evidenced from seed germination test and chlorophenol red assay in T-1 plants. Transgenic plants could be recovered after 8-10 weeks of cocultivation with Agrobacterium. An overall transformation frequency of 1.51% was achieved.
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