三种马铃薯病毒多重RT-PCR检测体系建立  

Establishment of multiplex RT-PCR system for detection of three potato viruses

作　　者：张威[1] 白艳菊[1] 罗丽环 范国权[1] 高艳玲[1] 申宇[1] 张抒[1] 孟宪欣[2] 

机构地区：[1]黑龙江省农业科学院植物脱毒苗木研究所,哈尔滨150086 [2]黑龙江省农业科学院作物育种研究所,哈尔滨150086

出　　处：《东北农业大学学报》2014年第5期13-18,共6页Journal of Northeast Agricultural University

基　　金：现代农业产业技术体系专项资金资助(CARS-10-P14);国家科技支撑项目(2012BAD06B02-02C2);黑龙江省自然科学基金项目(C201234);黑龙江省农业科技创新工程重点资助项目(2012ZD015)

摘　　要：马铃薯卷叶病毒(PLRV)、马铃薯Y病毒(PVY)和马铃薯S病毒(PVS)是马铃薯生产田中发生率较高、危害较严重的病毒,通常复合侵染。研究通过综合评价和筛选,确定每种病毒的特异性引物,对PCR部分各试剂用量和反应程序进行优化,在同一体系中对三种病毒复合侵染的马铃薯材料进行多重RT-PCR扩增,得到长度分别为336、447、729 bp的3条特异性条带,建立能同时检测PLRV、PVY和PVS病毒的多重RT-PCR检测体系。应用该体系对田间样品进行检测,检测结果特异性强、灵敏度高,表明该多重RT-PCR体系能够同时检测三种马铃薯病毒。The higher incidence and more serious viruses in potato field are potato leafroll virus（PLRV）, potato virus Y（PVY） and potato virus S（PVS）, and sometimes complexly infected. In this study, we found primers from Chinese and English documents, and national standards, through comprehensive evaluation and screening to determine the specific primers of each virus, at the same time, by optimization each reagent and response procedures of PCR, finally, the expected fragments of336 bp（PLRV）, 447 bp（PVY） and 729 bp（PVS） were successfully amplified by the multiplex RT-PCR system, and the multiplex RT- PCR detection system of PLRV, PVY and PVS were established. This system was useful for simultaneous detection of these three viruses from field samples. Using the system to detect field samples, the results were specificity and high sensitivity, indicate that the multiplex RT-PCR system can simultaneously detect three potato viruses.

关 键 词：马铃薯 PLRV PVY PVS 多重RT-PCR 检测 

分 类 号：S532[农业科学—作物学] Q944.57[生物学—植物学]