马铃薯4种病毒多重PCR检测体系的建立  

Establishment of multiplex PCR for four potato viruses

作　　者：庞博[1] 刘秀丽 张金文[1] 王蒂[1] 张俊莲[1] 

机构地区：[1]甘肃省作物遗传改良与种质创新重点实验室,甘肃农业大学农学院,兰州730070

出　　处：《植物保护》2015年第2期102-107,共6页Plant Protection

基　　金：国家自然科学基金(31260343);甘肃省科技重大专项(1102NKDA025)

摘　　要：我国马铃薯病毒主要有马铃薯Y病毒(PVY)、马铃薯X病毒(PVX)、马铃薯S病毒(PVS)、马铃薯卷叶病毒(PLRV),常发生复合侵染。根据GenBank中4种马铃薯病毒的外壳蛋白(coat protein,CP)基因全长设计引物,通过RT-PCR扩增得到4种病毒CP基因全长片段,测序结果显示序列同源性96%以上;针对4种病毒CP基因的保守序列分别设计引物,在一个PCR体系中同步对4种病毒进行扩增,得到421、202、516、330bp的特异性条带,优化建立了能同步检测PVY、PVX、PVS和PLRV的多重RT-PCR检测体系。检测结果证明优化后的多重RT-PCR体系能在田间样品中快速、高效地检测出4种病毒。Potato virus diseases are mainly caused by complex infection of Potato virus Y（PVY）,Potato virus X（PVX）,Potato virus S（PVS）,and Potato leaf roll virus（PLRV）.The primers for PVY,PVX,PVS and PLRV were designed based on the complete sequences of their coat protein（CP）genes and the sequence analysis of amplified fragments,which indicated that the four virus fragments shared at least 96 percent homology with other related viruses.The four pairs of specific primers based on the highly conserved sequences of the CP genes of the four viruses were designed.The target fragment sizes of 421 bp（PVY）,202 bp（PVX）,516 bp（PVS）and330 bp（PLRV）were amplified using multiplex RT-PCR.The optimized multiplex RT-PCR provides a quick and high efficiency method for simultaneous detection of the four viruses from field samples.

关 键 词：分子检测 多重RT-PCR 马铃薯病毒 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]