齿兰环斑病毒与建兰花叶病毒分子检测研究  

Molecular detection of Odontoglossum ringspot virus and Cymbidium mosaic virus

作　　者：闻伟刚[1] 谭钟[1] 崔俊霞[1] 陈先锋[1] 

机构地区：[1]宁波出入境检验检疫局,宁波315012

出　　处：《植物保护》2008年第6期65-69,共5页Plant Protection

基　　金：宁波出入境检验检疫局科技项目(甬K042005)

摘　　要：齿兰环斑病毒(Odontoglossum ringspot virus,ORSV)与建兰花叶病毒(Cymbidium mosaic virus,Cy MV)是严重危害兰科植物的两种主要病毒。本研究根据病毒外壳蛋白基因设计特异性引物,应用ELISA、普通RT-PCR、巢式RT-PCR和免疫捕获RT-PCR4种方法进行了检测研究与比较。结果表明:普通RT-PCR与ELISA方法检测灵敏度相当;巢式RT-PCR检测灵敏度要比普通RT-PCR与ELISA方法高出104倍以上;免疫捕获RT-PCR检测灵敏度介于普通RT-PCR和巢式RT-PCR之间。采用巢式RT-PCR方法对我国台湾进境的蝴蝶兰植株样本检测,1号样本出现与阳性对照一致的特异条带。双向测序分析,扩增产物序列与ORSV外壳蛋白基因具有100%的同源性,表明1号蝴蝶兰样本携带ORSV。Odontoglossum ringspot virus （ORSV） and Cymbidium mosaic virus （CyMV） are the two major types of viruses which infect Orchidaceous plants severely. In this study, based on the previously reported viral coat protein （CP） gene sequences, specific primers for ORSV and CyMV were designed, and four methods, including ELISA, RT-PCR, nested RT-PCR and IC-RT-PCR were performed for molecular detection of the two viruses. The results showed that the sensitivity of RT-PCR assay was equal to that of ELISA, but nested RT-PCR assay showed 10000 times higher in sensitivity than RT-PCR and ELISA, while the sensitivity of IC-RT-PCR was intermediate between RT-PCR and nested RT-PCR. Phalaenopsis that were imported from Taiwan of China were tested by nested RT- PCR, and a specific DNA fragment with the same size as positive control was obtained from sample 1. The sequencing results showed that its identity with the reported ORSV CP gene was 100%, which indicated that the Phalaenopsis sample 1 was infected with ORSV.

关 键 词：齿兰环斑病毒 建兰花叶病毒 巢式RT—PCR 免疫捕获RT-PCR 检测灵敏度 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治]