TaqMan实时荧光定量PCR检测茶黄蓟马  

TaqMan real-time fluorescent quantitative PCR for identification of Scirtothrips dorsalis Hoods

作　　者：李茵 刘英 双巧云[3] 彭徽 黄丽莉 Li Yin;Liu Ying;Shuang Qiaoyun;Peng Hui;Huang Lili(Department of Ecology and Environment,Yuzhang Normal University,Nanchang 330103,China;Technology Centre of Nanchang Customs District;Jiangxi Sericulture and Tea Research Institute)

机构地区：[1]豫章师范学院生态与环境系,江西南昌330103 [2]南昌海关技术中心 [3]江西省蚕桑茶叶研究所

出　　处：《植物检疫》2021年第6期43-47,共5页Plant Quarantine

基　　金：江西省重点研发计划一般项目(S2020ZPYFB0042)。

摘　　要：传统形态学方法难以区分茶黄蓟马与其相似种,特别在幼虫等未成熟阶段,本研究基于茶黄蓟马ITS2基因序列设计合成了特异性引物和TaqMan探针,建立了一种基于TaqMan实时荧光定量PCR技术的茶黄蓟马快速检测方法。结果表明:制作的标准曲线有良好的线性关系;对包括茶黄蓟马在内的茶园常见蓟马进行检测,能够特异性地检测到茶黄蓟马的荧光信号,而其他蓟马没有荧光信号;灵敏度检测结果显示可检测浓度为1690拷贝/μL的靶标目的片段DNA。该方法具有较强的特异性、较高的灵敏度,为茶黄蓟马的早期监测和口岸快速检疫提供了高效的检测鉴定手段。It is difficult to distinguish Scirtothrips dorsalis from its similar species,especially at immature stages such as larvae,by traditional morphological methods,so we designed and synthesized specific primers and TaqMan probes based on the specificity of ITS2 gene sequence of S.dorsalis,and established a rapid detection method based on TaqMan real-time fluorescence quantitative PCR.The results showed that the standard curve had an excellent linear relationship,and the real-time fluorescence quantitative PCR method could specifically detect the fluorescence signal of S.dorsalis from common tea thrip species,but it had no detection signal for other thrips.The sensitivity test showed that the target DNA fragment could be detected at a concentration of 1690 copies per microliter.The method has strong specificity and sensitivity,which provides an efficient detection and identification method for early monitoring and rapid quarantine of S.dorsalis at ports.

关 键 词：茶黄蓟马 实时荧光定量PCR TAQMAN探针 鉴定 

分 类 号：S41[农业科学—植物保护]