二重RT-PCR快速检测马铃薯病毒的方法

本研究采用传统的蛋白酶K法和病毒RNA简易浸提法,从马铃薯块茎、茎干、叶梗、叶片中提取马铃薯X病毒,马铃薯Y病毒,马铃薯A病毒及马铃薯卷叶病毒RNA,并设计了4种马铃薯病毒引物,优化了二重RT-PCR反应条件,可以同步扩增出上述4种病毒,扩增产生的靶带分别为562bp(PVX)、480bp(PVY)、336bp(PLRV)、255bp(PVA).应用病毒RNA简易制样技术和优化的二重RT-PCR反应条件,可以同步快速检测田间自然感染的马铃薯病毒,此研究还可适合于检测马铃薯脱毒种薯及试管苗,对马铃薯病毒病早期监测有一定的作用.

Simultaneous detection of potato viruses with Duplex RT-PCR

The protein K and simple virus RNA-release methods we re developed for extracting potato viruses(PVX,PVY,PLRV,PVA)from potato tuber s,stems,petioles and leaves. The primers for the viruses were designed and the duplex RT-PCR system was developed,which could detect PVY,PLRV simultaneousl y or PVX,PVA simultaneously and the amplification fragments of 562bp(PVX),480b p(PVY),336bp(PLRV),225bp(PVA) were generated respectively. The virus RNA-rele ase method and the duplex RT-PCR were also suitable for detection the viruses o f pro-potato-seed both from tissue cultural factory and from naturally field g rowing potato plant.