香蕉3种病毒的多重PCR检测体系  

Development of a Multiplex PCR Protocol for the Detection of Three Viruses in Banana

作　　者：谭小勇[1] 阮小蕾[1] 吴丽婷[1] 翟国会[1] 李华平[1] 

机构地区：[1]华南农业大学植物病毒研究室,广州510642

出　　处：《园艺学报》2010年第5期829-834,共6页Acta Horticulturae Sinica

基　　金：国家公益性行业科研专项(nyhyzx07-29);国家香蕉产业技术体系项目(nycytx-33-06);广东省产业化推进项目专项(2009B020201011)

摘　　要：根据GenBank中已发表的香蕉束顶病毒(Banana bunchy top virus,BBTV)﹑黄瓜花叶病毒(Cucumber mosaic virus,CMV)和香蕉线条病毒(Banana streak virus,BSV)的基因序列,找出保守序列分别设计出特异引物,在建立各种病毒单项PCR技术的基础上,通过优化多重PCR反应条件,建立了能同时检测3种病毒的多重PCR检测体系。该体系可同时扩增BBTV的615bp、CMV的480bp和BSV的945bp的特异片段。这些片段克隆测序结果表明,多重PCR扩增的不同病毒片段是特异和专化的,其核苷酸序列与相应的病毒基因片段的相似性都达91%以上。该体系的灵敏度测定结果表明,从相当于或大于10-2mg感病植物组织中均能检测到这3种病毒。Based on the establishment of the optimal PCR and RT-PCR for the detection of single virus,a multiplex PCR protocol for the detection of three main viruses （Banana bunchy top virus,BBTV;Cucumber mosaic virus,CMV;Banana streak virus,BSV） in banana is developed.Three specific fragments with molecular weights at 615 bp for BBTV,480 bp for CMV and 945 bp for BSV were simultaneously amplified in one PCR reaction system.The sequence analysis of amplified fragments indicates that three virus fragments shared above 91 percent identities with that of other relative viruses.It was showed that the sensitivity of this protocol equaled to be able to detect the three viruses at the level of 10-2 mg banana tissue.

关 键 词：香蕉 病毒 黄瓜花叶病毒 香蕉束顶病毒 香蕉线条病毒 多重PCR 

分 类 号：S668.1[农业科学—果树学]