山药潜隐病毒RT-LAMP快速检测方法的建立  

Establishment of RT-LAMP rapid detection method for yam latent virus

作　　者：王凤丽 王飞[2] 鲁传涛[2] 高素霞[2] 刘玉霞[2] 文艺[2] 杨瑾 李雪梦 戚文平 刘国彬 张红瑞[3] 蔡丽[1] 秦艳红[2] WANG Fengli;WANG fei;LU Chuantao;GAO Suxia;LIU Yuxia;WEN Yi;YANG Jin;LI Xuemeng;QI Wenping;LIU Guobin;ZHANG Hongrui;CAI Li;QIN Yanhong(College of Plant Science and Technology,Huazhong Agricultural University,Wuhan 430070,China;Institute of Plant Protection,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;College of Agriculture,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]华中农业大学植物科学技术学院,武汉430070 [2]河南省农业科学院植物保护研究所,郑州450002 [3]河南农业大学农学院,郑州450046

出　　处：《植物病理学报》2023年第4期743-747,共5页Acta Phytopathologica Sinica

基　　金：国家中药材产业技术体系(CARS-B21);河南省中药材产业技术体系(Z2019-13-01);河南省农业科学院新兴学科发展专项(2020XK05、2021XK02、2022XK07)。

摘　　要：0引言山药(Dioscorea oppositifolia L.)是薯蓣科薯蓣属植物,可作为药用或食用材料[1],广泛分布于全球热带及亚热带地区,在我国河南、山东、江苏、广西和江西等省份都有种植。山药生产中以块茎无性繁殖方式为主,导致病毒病发生严重[2]。侵染山药的病毒主要包括马铃薯Y病毒属(Potyvirus)[3]、杆状DNA病毒属(Badnavirus)[4]、香石竹潜隐病毒属(Carlavirus)[5]以及蚕豆病毒属(Fabavirus)[6]的一些病毒。Yam latent virus is an important virus that infects yam.It can cause mosaics and chlorosis on yam leaves and affect both the quality and yield.In this study,6 specific primer pairs were designed according to the cp gene sequence of yam latent virus(YLV),and a reverse transcription loop-mediated isothermal amplification(RT-LAMP)assay with strong specificity and high sensitivity was established.The target product was amplified at 50-65℃for 1 hour,and the optimal reaction temperature was 58-65℃.The specificity result showed that this method was able to specifically identify YLV from the other three RNA viruses infecting yam.The sensitivity test showed that the detection limit of the LAMP method was 5.08×101 copies·μL^(-1).The sensitivity of this method was 100 times that of the conventional PCR.The results indicated that this method can accurately detect YLV from the field samples.The RT-LAMP established in this study is sensitive,specific and reliable,thus making it suitable for the rapid detection of YLV.

关 键 词：YAM yam latent virus reverse transcription loop-mediated isothermal amplification rapid detection 

分 类 号：S432.44[农业科学—植物病理学]