文心兰3种主要病毒多重RT-PCR检测体系的建立  

Multiplex RT-PCR for Simultaneous Detection of Three Viruses in Oncidium

作　　者：樊荣辉[1] 黄敏玲[1] 钟淮钦[1] 吴建设[1] 罗远华[1] 

机构地区：[1]福建省农业科学院作物研究所/福建省农业科学院花卉研究中心/福建省特色花卉工程技术研究中心,福建福州350013

出　　处：《福建农业学报》2015年第7期697-700,共4页Fujian Journal of Agricultural Sciences

基　　金：福建省科技重大专项(2012NZ0002);福建省自然科学基金(2014J01113);福建省林业科学研究项目(闽林种站[2013]42号);福建农业科学院导师制青年人才创新基金(2013DQA-4)

摘　　要：根据GenBank中已发表的建兰花叶病毒(Cymbidium mosaic virus,CyMV)、齿兰环斑病毒(Odontoglossum ringspot virus,ORSV)和菜豆黄花叶病毒(Bean yellow mosaic virus,BYMV)外壳蛋白(CP)基因序列保守区域分别设计特异性引物,通过优化多重PCR反应条件,建立能同时检测文心兰3种病毒的多重PCR检测体系。该体系能够一次扩增出CyMV、ORSV和BYMV的特异片段,其大小分别是551、325和212bp。测序结果表明,3种病毒序列与相应的参考序列相似性均达98%以上。灵敏度测定结果表明,从相当于或大于10-2 mg的感病植物组织中能够检测到这3种病毒。By using specific primers designed on the basis of conserved sequences of coat protein gene ,a multiplex PCR protocol for the detection of 3 commonly found disease‐causing viruses ,i .e .,Cymbidium mosaic (CyMV) , Odontoglossum ringspot (ORSV) and bean yellow mosaic (BYMV) ,in Oncidium was developed .Three specific fragments were simultaneously amplified in a single PCR reaction system .Their molecular weights were determined to be 571 bp for CyM V ,325 bp for ORSV ,and 212 bp for BYM V .Sequence analysis indicated that the 3 viruses were at least 98% homogenous with their respective reference viruses .The sensitivity test on this newly developed methodology showed that it was capable of accurately detecting the 3 viruses simultaneously in a bulbous iris tissue at the level of 10-2 mg .

关 键 词：文心兰 病毒 建兰花叶病毒 齿兰环斑病毒 菜豆黄花叶病毒 多重RT-PCR 

分 类 号：S682[农业科学—观赏园艺]