番茄环纹斑点病毒核衣壳蛋白N的RT-LAMP检测方法的建立  

Establishment of a Reverse Transcription Loop￣Mediated Isothermal Amplification(RT￣LAMP)Method for Detecting Nucleocapsid Protein N of Tomato Zonate Spot Virus

作　　者：罗海燕 李恒 陆承聪 魏辉[1] 郑雪[2] 陈勇[1] Luo Haiyan;Li Heng;Lu Chengcong;Wei Hui;Zheng Xue;Chen Yong(Institute of Plant Protection,Fujian Academy of Agricultural Sciences/State Key Laboratory for Ecological Pest Control of Fujian and Taiwan Crops/Fuzhou Scientific Observing and Experimental Station of Crop Pests,Ministry of Agriculture and Rural Affairs,Fuzhou 350013,China;Institute of Biotechnology and Germplasm Resources,Yunnan Academy of Agricultural Sciences,Kunming 650205,China)

机构地区：[1]福建省农业科学院植物保护研究所/闽台作物有害生物生态防控国家重点实验室/农业农村部福州作物有害生物科学观测试验站,福建福州350013 [2]云南省农业科学院生物技术与种质资源研究所,云南昆明650205

出　　处：《山东农业科学》2023年第11期176-180,共5页Shandong Agricultural Sciences

基　　金：福建省科技计划项目(2023J06040,2022R1024006);福建省农业科学院科技项目(CXTD2021004-3,XTCXGC2021011,XTCXGC2021017)。

摘　　要：番茄环纹斑点病毒(Tomato zonate spot virus,TZSV)属正番茄斑萎病毒属(Orthotospovirus),严重影响番茄、辣椒等作物的产量及经济价值。本研究根据TZSV的核衣壳蛋白N的基因序列设计逆转录环介导等温扩增(reverse transcription loop-mediated isothermal amplification,RT-LAMP)反应引物,建立了TZSV的RT-LAMP检测方法。结果显示,该方法能够特异扩增TZSV,与侵染辣椒的番茄斑萎病毒和烟草花叶病毒不发生反应;RNA最低检测限为0.01 pg/μL,灵敏度为普通逆转录PCR(RT-PCR)的10倍;田间样品检测应用结果表明,RT-LAMP扩增和可视化检测结果与RT-PCR一致。综上,本研究建立的RT-LAMP快速检测方法可有效应用于TZSV的田间检测。Tomato zonate spot virus(TZSV)is a phytopathogen of the genus Orthotospovirus,which cau-ses severe economic value and yield loss of tomato,pepper and other crops.In this study,the reverse tran-scription loop-mediated isothermal amplification(RT-LAMP)primers were designed originally using the gene sequence of the nucleocapsid protein N of TZSV,and then a specific method was established for the detection of TZSV by using RT-LAMP method.The results showed that this method could amplify TZSV specifically,and could not react with tomato spotted wilt virus and tobacco mosaic virus.The lowest detection limit of RNA was 0.01 pg/μL,and the sensitivity was 10 times of that of ordinary reverse transcription PCR(RT-PCR).The RT-LAMP method was applied to detect TZSV in field samples,and the results of RT-LAMP amplification and visual inspection were consistent with those of RT-PCR.In conclusion,the RT-LAMP rapid detection method established in this study could be effectively applied to the field detection of TZSV.

关 键 词：番茄环纹斑点病毒 核衣壳蛋白N 逆转录环介导等温扩增技术 检测 

分 类 号：S432.41[农业科学—植物病理学]