关键词: 太子参, 芜菁花叶病毒, 蚕豆萎蔫病毒, 双重RT-PCR

Abstract:

Ahigh incidenceof both Turnip mosaic virus（TuMV）and Broad bean wilt virus （BBWV） in cultivated Pseudostellaria heterophylla in different producing areas causes serious damage. A duplex RT-PCR method was developed for simultaneous and rapid detection of those 2 viruses. Degenerate primers were designed according to the conserved sequence regions of coat protein（CP）genes from GenBank. Duplex RT-PCR detection system was established and optimized based on single RT-PCR. The optimal parameters for the duplex RT-PCR were 35 PCR cycles，0.4 μmol · L^-1 for the concentration，and an annealing temperature at 51 ℃. of each of the primers Sensitivity analysis revealed that viruses could be detected from 10⁴-fold dilution of the first strand cDNA. Duplex RT-PCR established in this study was used to detect 2 viruses in 7 samples of cultivated P. heterophylla and 4 samples of virus-free test-tube plantlets. The results proved that it could be used for rapid detection of TuMV and BBWV  simultaneously in P. heterophylla with high stability，accuracy and sensitivity.

Key words: Pseudostellaria heterophylla, TuMV, BBWV, duplex RT-PCR