二重RT-PCR快速检测马铃薯病毒的方法  

Simultaneous detection of potato viruses with Duplex RT-PCR.

作　　者：袁青[1] 殷幼平[1] 王中康[1] 黄振霖 

机构地区：[1]重庆大学基因工程研究中心,400044 [2]重庆市植保植检站

出　　处：《植物检疫》2005年第3期135-138,共4页Plant Quarantine

摘　　要：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)、3 3 6bp(PLRV)、2 55bp(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。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.

关 键 词：PCR快速检测 RT-PCR 马铃薯卷叶病毒 马铃薯X病毒 马铃薯Y病毒 马铃薯A病毒 马铃薯病毒病 反应条件 马铃薯块茎 RNA 制样技术 自然感染 脱毒种薯 浸提法 蛋白酶 试管苗 优化 同步 茎干 田间 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]