油菜花叶病毒RT-LAMP快速检测方法的建立  

Establishment of RT-LAMP method for rapid detection ofyoucai mosaic virus

作　　者：秦艳红[1] 王凤丽 文艺[1] 赵正伟 高素霞[1] 张德胜[1] 李绍建[1] 刘永康 刘玉霞[1] 鲁书豪 王飞[1] 鲁传涛[1] QIN Yanhong;WANG Fengli;WEN Yi;ZHAO Zhengwei;GAO Suxia;ZHANG Desheng;LI Shaojian;LIU Yongkang;LIU Yuxia;LU Shuhao;WANG Fei;LU Chuantao(Institute of Plant Protection,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;Wenxian Agriculture Science Research Institute,Wenxian 475004,China)

机构地区：[1]河南省农业科学院植物保护研究所,郑州450002 [2]温县农业科学研究所,温县475004

出　　处：《植物病理学报》2024年第4期829-834,共6页Acta Phytopathologica Sinica

基　　金：国家中药材产业技术体系(CARS-21);河南省中药材产业技术体系(HARS-22-11-Z1);河南省农业科学院基本科研业务费项目(2023ZC051)。

摘　　要：油菜花叶病毒(youcai mosaic virus,YoMV)是侵染地黄和山药的重要病毒之一,能导致中药材的产量下降及品质降低。本研究基于逆转录环介导恒温扩增技术(reverse transcription loop-mediated isothermal amplification,RT-LAMP),建立了针对YoMV的快速、特异、灵敏的RT-LAMP检测方法。根据YoMV外壳蛋白(coatprotein,CP)的核苷酸序列设计了5条特异性引物,建立的RT-LAMP方法最适检测温度为60℃,反应时间为60min,该方法能特异性检测到YoMV,灵敏度是常规PCR的1000倍,可检测9.18×10^(2)拷贝·μL^(-1)模板,本研究建立的RT-LAMP检测技术为地黄和山药上YoMV的准确鉴定提供了快速、高效的方法。Youcai mosaic virus(YoMV)is one of the important viruses infecting Rehmannia glutinosa and Dioscorea oppositifolia.It decreases the yield and quality of these Chinese medicinal herbs.Based on reverse transcription loop-mediated isothermal amplification(RT-LAMP),a rapid,specific and sensitive method for detecting YoMV was developed.Five specific primers were designed according to the nucleotide sequence of YoMV coat protein(CP).The established RT-LAMP method for detecting YoMV was optimally performed at 60℃for 60 min.This method can detect YoMV specifically.The sensitivity was 1000 times higher than that of the conventional PCR,and it could detect 9.18×10^(2)copies·μL^(-1).The RT-LAMP method established in this study provided a rapid and efficient diagnostic tool for correct detection of YoMV on Rehmannia glutinosa and Dioscorea oppositifolia.

关 键 词：地黄 山药 油菜花叶病毒 逆转录环介导恒温扩增 快速检测 

分 类 号：S432.44[农业科学—植物病理学]