球根鸢尾3种主要病毒多重RT-PCR检测体系的建立  

Development of Multiplex RT-PCR for Simultaneous Detection of Three Viruses in Bulbous Iris

作　　者：樊荣辉[1,2,3] 黄敏玲[1,2,3] 钟淮钦[1,2,3] 吴建设[1,2,3] 林兵[1,2,3] 

机构地区：[1]福建省农业科学院作物研究所,福建福州350013 [2]福建省农业科学院花卉研究中心,福建福州350013 [3]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2期172-175,共4页Fujian Journal of Agricultural Sciences

基　　金：福建省科技计划重大专项(2010NZ0003;2012NZ0002);福建省财政专项--福建省农业科学院科技创新团队建设项目(CXTD-2-17;CXTD-2-1317);福建省科技计划项目--省属公益类科研院所基本科研专项(2014R1026-12)

摘　　要：根据GenBank中已发表的鸢尾轻花叶病毒(Iris mild mosaic virus,IMMV)、水仙潜隐病毒(Narcissus latent virus,NLV)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,建立能同时检测球根鸢尾3种病毒的多重PCR检测体系。该体系能够一次扩增出IMMV、NLV和BYMV的特异片段,其大小分别是770bp、266bp和186bp。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 （CP） gene ,a multiplex PCR protocol for the detection of three main viruses （Iris mild mosaic virus ,IMMV ;Narcissus latent virus ,NLV ;Bean yellow mosaic virus ,BYMV） in Bulbous Iris was developed. Three specific fragments were simultaneously amplified in one PCR reaction system. Their molecular weights were 770 bp ,266 bp and186 bp respectively. Sequence analysis indicated that the three virus sequences shared at least 98 percent identities with that of other relative viruses. The result showed that the sensitivity of this protocol equaled to be able to detect the three viruses at the level of 10-2 mg Bulbous Iris tissue.

关 键 词：球根鸢尾 鸢尾轻花叶病毒 水仙潜隐病毒 菜豆黄花叶病毒 多重RT-PCR 

分 类 号：S432[农业科学—植物病理学]