Detection of PVY, PVS and PLRV by Multiplex RT-PCR

Virus is the one of the plant pathogen which is harmful to the potatoes production. In order to rapid detect the main potatoes virus species of Yunnan Province, the PCR primers for potato virus Y (PVY), potato virus S (PVS), and potato leaf roll virus (PLRV) were designed and synthesized based on their coat protein gene sequences. Then the optimal system of triple RT-PCR detection system was established from annealing temperature, minimum detectable concentration, thermal cycling condition and best combination concentration of virus template with primers. The result showed that this optimized multiplex RT-PCR could amplify three viruses simultaneously, and the fragments were 480bp (PVY), 187bp (PVS) and 336bp (PLRV), respectively. This optimized multiplex RT-PCR reaction system was proved to be rapid, sensitive and simple to detect single or compound infection of the above potato virus and to early detection and control of potato virus diseases. This result will have an important practical effect on potato seedlings detoxification, trans-generational and high yield cultivation in the field in Yunnan Province.