双重RT-PCR法快速检测多种马铃薯病毒的研究  

Quick Detection of Potato Viruses by Duplex RT-PCR

作　　者：陈阳婷[1] 桑有顺[1] 冯焱[1] 陈涛[1] 

机构地区：[1]成都市农林科学院,成都温江611130

出　　处：《西南农业学报》2012年第1期179-182,共4页Southwest China Journal of Agricultural Sciences

基　　金：国家马铃薯产业技术体系成都综合试验站项目(CARS-10-ES17)

摘　　要：为建立快速、简便、灵敏度高、特异性强的RT-PCR检测马铃薯病毒技术,依据马铃薯X病毒、马铃薯S病毒、马铃薯A病毒以及马铃薯卷叶病毒的基因组RNA保守序列设计特异性引物对,从反转录和PCR等方面优化双重RT-PCR反应条件,同步扩增出上述四4种病毒,分别得到620(PVX)、435(PVS)、300(PVA)、222 bp(PLRV)大小的扩增片段。结果表明,反转录反应中引物浓度比例、dNTPs浓度和PCR反应中Mg2+浓度对双重RT-PCR检测多种马铃薯病毒的影响较大。优化的双重RT-PCR反应体系可以同步快速检测田间自然感染的马铃薯病毒,此方法还适合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。In order to establish a technique using RT-PCR to detect potato viruses,which was rapid,accurate and sensitive,the primers for PVX,PVS,PVA and PLRV fragments were designed based on their coat protein,and a duplex reverse transcription polymerase chain reaction（d-RT-PCR） was optimized from reverse transcription（RT） stage and polymerase chainreaction（PCR） stage.The optimized d-RT-PCR can amplify PVX,PVS,PVA and PLRV simultaneously,and the fragments were 620 bp（PVX）,435 bp（PVS）,300 bp（PVA）, 222bp（PLRV）.The result showed that antisense primer rations of two viruses,dNTPs concentration and Mg2＋ concentration had great impacts on the detection.The duplex RT-PCR was also suitable for viruses detection of propotato seed both from tissue cultural factory and from naturally field growing potato plant.

关 键 词：马铃薯 RT-PCR 病毒检测 

分 类 号：S436.32[农业科学—农业昆虫与害虫防治]