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
刘三军,郭卫东,张秋叶,何水涛,周厚成,张亚冰
摘要(Abstract):
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。
关键词(KeyWords): 葡萄卷叶病毒Ⅲ;RT-PCR;检测;克隆;序列分析
基金项目(Foundation): 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
作者(Author): 刘三军,郭卫东,张秋叶,何水涛,周厚成,张亚冰
Email:
DOI: 10.13925/j.cnki.gsxb.2002.01.005
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