马铃薯纺锤块茎类病毒RT-qPCR检测技术体系的建立  

Establishment of RT-qPCR Detection System for Potato Spindle Tuber Viroid

作　　者：邱彩玲[1] 范国权[1] 申宇[1] 高艳玲[1] 张威[1] 韩树鑫 张抒[1] 董学志[1] 马纪[1] 白艳菊[1] Qiu Cailing;Fan Guoquan;Shen Yu;Gao Yanling;Zhang Wei;Han Shuxin;Zhang Shu;Dong Xuezhi;Ma Ji;Bai Yanju(Potato Research Institute,Heilongjiang Academy of Agricultural Sciences/Supervision and Testing Center for Virus-Free Seed Potatoes Quality of Ministry of Agriculture and Rural Affairs(Harbin),Harbin 150086,Heilongjiang,China;Plant Protection Research Institure of Heilongjiang Academy of Land Reclamation Sciences,Harbin 150000,Heilongjiang,China)

机构地区：[1]黑龙江省农业科学院马铃薯研究所/农业农村部脱毒马铃薯种薯质量监督检验测试中心(哈尔滨),黑龙江哈尔滨150086 [2]黑龙江省农垦科学院植物保护研究所,黑龙江哈尔滨150000

出　　处：《作物杂志》2020年第3期79-84,共6页Crops

基　　金：黑龙江省农业科学院科研项目(2019JJPY013,2019CGJL004);黑龙江马铃薯产业技术协同创新推广体系。

摘　　要：马铃薯纺锤块茎类病毒(PSTVd)是马铃薯生产中的一种重要病害,目前主要通过使用脱毒种薯及隔离措施来防控,探索高效、灵敏、特异性强的检测技术对于防治该病害具有重要意义。设计了3组引物,1条探针,从3组引物中筛选出最优组合,并以他们为引物,以PSTV 251T为探针,利用5′FAM、3′TAMRA标记,建立了PSTVd的RT-qPCR检测技术体系。利用该检测体系成功地检测了22份马铃薯样本。与灵敏度相对较高的RT-PCR技术相比,该检测技术体系的检测灵敏度又提升了100~1000倍。Potato spindle tuber viroid(PSTVd)is an important disease in potato production.The disease currently relies mainly on the use of virus-free seed potatoes and isolation measures to control.For PSTVd control,it is very important to explore a detection technology with high efficiency,sensitivity and specificity.In this study,three pairs of primers and one probe were designed and the optimal combination was selected from them.By use of these primer pairs and the probe PSTV 251T,marked with 5'FAM and 3'TAMRA,a PSTVd RT-qPCR detection system was established and 22 potato samples were tested successfully using this system.Compared with the RT-PCR technology with relatively high sensitivity,the sensitivity of this RT-qPCR system was increased 100-1000 times.

关 键 词：马铃薯纺锤块茎类病毒(PSTVd) RT-QPCR 检测技术 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]