DEVELOPMENT OF MOLECULAR RESISTANCE IN POTATO AGAINST POTATO LEAF ROLL VIRUS AND POTATO VIRUS Y THROUGH AGROBACTERIUM-MEDIATED DOUBLE TRANSGENESIS

Abstract

Potato leafroll virus (PLRV) and potato virus Y (PVY) are the two major viral problems for the potato production all over the world. Transgenic approaches involving the expression of viral genes are being developed to provide protection for plants against viral diseases. The purpose of this study was to develop double transgenic plants of potato using PLRV replicase and PVY coat protein genes tandemly placed in a single T-DNA transformant through Agrobacterium-mediated transformation. A total of 17 lines of putative transformants of potato cv. Desiree were generated from kanamycine resistant calli originated from co-inoculation of separate Agrobacterium cultures containing PVY CP and PLRV replicase genes. Shoots were excised and cultured onto shoot medium containing 250mg/L cefotaxime and 50mg/L kanamycin sulfate in test tubes. Polymerase chain reaction (PCR) analysis was conducted of 39 plants of 16 transformed lines using primers each of PVY CP and PLRV-replicase genes; 10 plants of 8 lines and 7 plants of 6 lines showed presence of of PVY CP and PLRV-replicase genes, respectively. However