Abstract: This investigation highlights a reproducible method of Agrobacterium-mediated genetic transformation developed for V. aconitifolia cv BMB-43. Efficient plant regeneration via somatic embryogenesis has been developed using leaf explants of in vitro derived seedlings. The plasmid contained a bi-functional fusion gene conferring both neomycin phosphotransferase gene (nptII) and β-glucuronidase (GUS) gene (gus A) interrupted with an intron, both driven by the Cauliflower Mosaic Virus (CAMV) 35S promoter. Using a series of tissue culture media for the establishment of embryogenic cultures, co-cultivation, selection, maturation and rooting, we recovered transgenic mothbean plants. Expression of positive histochemical GUS activity (51.0%) in the transgenic seedlings was observed. The integration of the stable transgenes (65.3%) in the plant genomes was shown by means of PCR amplification of these genes from plant genomic DNA and Southern blot hybridization with gene-specific probes. This method allows high-efficiency production of transgenic plants in mothbean.