Agrobacterium-mediated transformation of tomato (Lycopersicon esculentum var. Pusa ruby) with coat-protein gene of Physalis mottle tymovirus

Transgenic tomato plants harboring the coat protein (CP) gene of Physalis m ottle tymovirus (PhMV) were obtained through Agrobacterium-mediated genetic transformation. The cotyledonary leaves were transformed by cocultivation with Agrobacterium strain LBA4404 carrying the binary vector pBI 121. The C P gene of PhMV was cloned into the binary vector by deleting the GUS gene. Cotyledonary explants were cultured bn the regeneration medium supplemented with 50 mg/L kanamycin and 400 mg/L cefotaxime (MS1). Shoots regenerated v ia callus. Putative transformants were transferred to a rooting medium cont aining 25 mg/L kanamycin (MS2), where successful rooting was obtained. Tran sgenic nature of the transformants was checked by PCR and Western analyses. The T-1 progenies segregate into a 3:1 ratio. Southern analysis confirmed the integration of CP gene into the tomato genome of T-1 progenies. The tra nsgenic tomato plants were further challenged with PhMV and exhibited delay in symptom development, thus confirming the establishment of transgenic pl ants with partial resistance to the virus.