马铃薯PVY和PLRV病毒的TC-RT-PCR检测  

Detection of Potato Virus Y and Potato Leafroll Virus Using Tube Capture RT-PCR

作　　者：郑叶叶 韩磊[1] 迟胜起[1] 张剑峰[1] 

机构地区：[1]青岛农业大学农学与植物保护学院,山东青岛266109

出　　处：《中国马铃薯》2016年第5期296-301,共6页Chinese Potato Journal

基　　金：山东现代农业产业技术体系薯类创新团队项目(SDAIT-16-06);国家科技支撑计划项目(2011BAD32B03-3)

摘　　要：传统的RT-PCR技术检测病毒需提取总RNA,RNA容易降解。利用试管捕捉反转录扩增(Tube cap-ture RT-PCR,TC-RT-PCR)方法检测了PVY和PLRV 2种病毒,实现了不需提取总RNA也可在同一反应中同时检测2种病毒。根据已报道的引物用TC-RT-PCR的方法对PVY和PLRV的外壳蛋白基因进行了检测。结果表明,TC-RT-PCR能够成功的检测出感染PVY或PLRV以及2种病毒共同侵染的样品,扩增产物序列长度均与设计片段的长度相符,分别为781和364 bp,2种病毒扩增产物的测序结果同Gene Bank中已知的序列比对后的同源性均高达97%以上。该技术为单独或复合感染的马铃薯病毒的检测提供了更加方便、高效的方法。同时测得试验PVY病毒样本属于PVYNW株系。To detect the virus using traditional RT-PCR technique needs extraction of RNA which degrades easily.Using tube capture reverse transcription amplification（Tube capture RT-PCR, TC-RT-PCR） method to detect two viruses, potato virus Y（PVY）, and potato leafroll virus（PLRV）, can be done in the same reaction system without extraction of total RNA. According to the reported primers, PVY and PLRV coat protein genes were detected with the method of TC-RT-PCR. The results showed that TC-RT-PCR successfully detected the samples infected with PVY or PLRV alone and coinfected with the two viruses, and the length of the amplified PVY and PLRV products was consistent with the length of the design fragments, 781 bp and 364 bp, respectively. Sequencing of the two virus products with the sequence alignment in Gene Bank homology were above 97%. The technique would provide a more convenient and efficient method for detecting potatoes for simple or complex virus infection. At the same time, the experiment found that the resulting PVY virus samples belonged to PVYNWstrain.

关 键 词：PVY PLRV 试管捕捉RT-PCR 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]