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当前位置: 首页 > 文章 > 黄瓜花叶病毒分离物核酸酶保护试验(RPA)方法的株系分化研究 植物病理学报 2005 (1) 47-51
Position: Home > Articles > Strain differentiation analysis of Cucumber mosaic virus with ribonuclease protection assay(RPA) Acta Phytopathologica Sinica 2005 (1) 47-51

黄瓜花叶病毒分离物核酸酶保护试验(RPA)方法的株系分化研究

作  者：
田兆丰;裘季燕;刘伟成;李永丹
单  位：
中国农业大学植物保护系;北京市农林科学院植保环保所
关键词：
黄瓜花叶病毒(CMV);株系分化;核酸酶保护试验
摘  要：
采用黄瓜花叶病毒((CMV)亚组Ⅰ株系Fny—CMV及亚组Ⅱ株系Ls—CMV的RNA2的特定序列片段的cDNA克隆,体外转录,同时掺入32P标记制备负链RNA探针,再与纯化的甜椒上的CMV中国分离物的RNA进行杂交,检测其与探针之间的同源性。共检测样品分离物3份。试验结果表明:河南新乡和北京密云的CMV甜椒分离物与Fny-CMV的核苷酸有高度同源性,隶属于Fny-CMV为代表的亚组Ⅰ株系。来自福建的样品与亚组Ⅱ的Ls-CMV株系有高度同源性,隶属于 CMV亚组Ⅱ株系。本试验同时利用源于我国CMV亚组Ⅰ的K株系的RNA2两个EcoR Ⅰ位点间1657-2125 nt的核苷酸序列为探针,同样与以上3份CMV中国分离物进行RNA杂交,进一步比较分析了这几个分离物与我国亚组Ⅰ的K-CMV 株系的关系,证明了我国CMV存在亚组与株系分化。
译  名：
Strain differentiation analysis of Cucumber mosaic virus with ribonuclease protection assay(RPA)
作  者：
TIAN Zhao-feng, QIU Ji-yan, LIU Wei-cheng, LI Yong-dan (Institute of Plant & Environment Protection, Beijing Academy of Agriculture & Forestry Sciences, Beijing 100089, China; Department of Plant Protection,China Agriculture University,Beijing 100094,China)
关键词：
Cucumber mosaic virus (CMV); strain differentiations; ribonuclease protection assay
摘  要：
cDNA sequences spanning 1209 nt to 1626 nt of Fny-CMV RNA2(a subgroup Ⅰ strain) and 2002 nt to 2433 nt of Ls-CMV RNA2(a subgroup Ⅱ strain) were used for probe preparation. The 32P-labeled negative strand RNAs were prepared by transcription in vitro. Then 3 Chinese CMV isolates from pepper were purified and hybridized with RNA-probes respectively. The results showed that Beijing and Henan isolates share high sequence identities with Fny-CMV, belonging to CMV subgroup Ⅰ strain. Another isolate from Fujan province was belonged to CMV subgroup Ⅱ strain. Meanwhile, the purified RNAs of the 3 Chinese isolates were hybridized with a transcription probe complementary to the fragment flanking 1657nt to 2125nt of K-CMV RNA2(a subgroup Ⅰ strain originated from China). The differentiations of Chinese isolate were analyzed according to the results.

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