RT-PCRによるジャガイモ休眠塊茎からの4種ウイルスの検出

抄録

Several primer pairs were studied for detecting Potato virus Y (PVY), Potato leafroll virus (PLRV), Potato virus X (PVX), and Potato virus S (PVS) in dormant potato tubers by reverse transcriptionpolymerase chain reaction (RT-PCR). The specific c DNA of each virus was amplified well by the following primer pairs: PVYCP6P (5'-CGTCCAAAATGAGAATGCC-3') and PVYCP 6 M (5'-TCTTGTGTACTGATGCCAC-3') for PVY, PLRVCP9P (5'-CGCTCAAGAAGAACTGGAG-3') and PLRVCPE1M (5'-CCGAATTCCTATTTGGGGTTTTGCAA-3') for PLRV, X1P (5'-TCCTTATTCCAACGGCATC-3') and X1M (5'-ATCTAGGCTGGCAAAGTCG-3') for PVX, and S7P (5'-TTCCCAACAGGCGCAGTG-3') and S 2 M (5'-CTAAACGGTCTGCCTTCAT-3') for PVS. The amplified fragments of PVY, PLRV, PVX and PVS were 577bp, 470bp, 337bp, and 426bp, respectively. The minimum detectable purified viral RNA by RT-PCR was 100 fg for PVY, 1 pg each for PLRV and PVX, and 10 pg for PVS. Simple extraction without using organic solvent was studied and found to extract nucleic acids containing virus RNA in dormant potato tubers in about 2 hr.