葡萄卷叶病毒Ⅲ RT-PCR检测技术研究  

Detection of Grapevine Leafroll-associated Virus Ⅲ by Reverse Tran-scription-polymerase Chain Reaction

作　　者：刘三军[1] 郭卫东[1] 张秋叶[1] 何水涛[1] 周厚成[1] 张亚冰[2] 

机构地区：[1]中国农业科学院郑州果树研究所,河南郑州450009 [2]西北农林科技大学,陕西杨凌712100

出　　处：《果树学报》2002年第1期15-18,共4页Journal of Fruit Science

基　　金：中国农业科学院院长基金资助项目(编号9921-9)及河南省重大科技攻关项目"苹果;葡萄;草莓脱毒快繁及产业化应用"资助项目

摘　　要：利用ＲＴ－ＰＣＲ技术,对葡萄卷叶病毒株系Ⅲ进行了检测。采用的技术流程为:通过提取葡萄叶片或韧皮部总ＲＮＡ,获得病毒的ＲＮＡ,反转录合成ｃＤＮＡ,经ＰＣＲ扩增,获得病毒ｃＤＮＡ的特征片段,并进行了序列分析。对提取的总ＲＮＡ进行了完整性和纯度检测,确定了良好的ＲＮＡ获得方法。结果表明,通过ＲＴ－ＰＣＲ扩增的片段序列与病毒源序列的同源性高达９９．３%,说明采用本研究确定的方法检测葡萄卷叶病毒株系Ⅲ结果可靠。An efficient procedure for the extraction of high-quality RNA from grape leaf and bark is described.A stable proce-dure for detection of grapevine leafroll-associated virus type Ⅲ was set up by the Reverse Transcription-Polymerase ChainReaction.The main technique is the method of the extraction of total RNA from grapevine leaves or bark.A 300 bp fragmentwas acquired by RT-PCR and the sequence of this fragment was aligned with the original sequence of grapevine leafroll-as-sociated virus type Ⅲ.The result showed that the identity of two sequence reach 99.3%.This indicated that the detection resultwas reliable.

关 键 词：葡萄 卷叶病毒Ⅲ RT-PCR 检测 克隆 序列分析 

分 类 号：S436.631.1[农业科学—农业昆虫与害虫防治] S432.41[农业科学—植物保护]