Abstract： The present research aims to develop transgenic potato plants resistance to PVX and PVY. RT-PCR was employed to clone a 267 bp fragment X from PVX-cp gene and a 294 bp fragment Y from PVY-cp gene. The two cloned fragment were ligated to form a fused gene XY. Then the fused gene XY was inserted in both sense and antisense orientation into vector pHANNIBAL to get a vector named pHIV. A Not Ⅰ fragment from pHIV containing a complete reading framework of an inverted repeat of fused gene XY was inserted to pART27 to get a vector pARIV which aimed to silence PVX-cp gene and PVY-cp gene simultaneously. Potato callus from elite potato cultivar Favorita was transformed via Laser microbeam puncture technique and 14 transgenic plants were regenerated. Southern blot analysis shows that there are 2-4 copies of fused XY gene in the transgenic plants. Virus infection indicates that 11 of the transgenic plants are immune to the infection of PVX、PVYo or PVX and PVYo , while the other three can be infected but virus concentration is much lower than that of untransformed plants infected by the same virus or viruses. This research would be helpful to develop transgenic plants resistance to multiple viruses using RNAi technology. It also suggests that Laser microbeam puncture technique is an efficient method to deliver RNAi vector into plants.