The binary plant cloning vector pCAMBIA1305.2 carrying the MYMV Replicase (Rep) gene was constructed and transformed into a competent Agrobacterium tumefaciens, strain LBA4404 and was used for co-cultivation of mungbean varieties, KKM-3, IC-39340-1, China mung and LM-1668 for imparting mungbean yellow mosaic virus (MYMV) resistance. The putative transformants (To generation) were selected on shooting media (SM) supplemented with 5 mgl-1 BAP, 50 mgl-1 hygromycin and 250 mgl-1 cefotaxime. The multiple shoots were inoculated on shoot elongation media (SEM) with 0.3 mgl-1 zeatin and 2.5 mgl-1 BAP for elongation and rooted on rooting media (RM), comprising half MS supplemented with 0.2 mgl-1 NAA. The GUS assay and genomic PCR analysis of the To transformants revealed, four positive putative transgenic lines for Rep gene specific primers and had a positive correlation with vector specific hptII primers. Likewise, VirG amplification revealed no Agrobacterium contamination in the apoplast of allthe putative To transformants.