Css. Vidya et al., Agrobacterium-mediated transformation of tomato (Lycopersicon esculentum var. Pusa ruby) with coat-protein gene of Physalis mottle tymovirus, J PLANT PHY, 156(1), 2000, pp. 106-110
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.