新疆小西葫芦黄化花叶病毒的分子鉴定与序列分析

在新疆石河子地区采集具有黄化、花叶和疱斑等症状的多种葫芦科作物,分别提取总RNA,利用小西葫芦黄化花叶病毒(Zucchini yellow mosaic virus,ZYMV)特异性引物进行RT-PCR扩增,其中来自黄瓜、甜瓜和丝瓜的模板RNA上可以得到1185bp片段。将扩增得到的片段克隆到pMD18-T上,序列分析结果表明:该片段含有1185个核苷酸,包含完整CP基因。与ZYMV Florida分离物、California、韩国KR-PA分离物、新加坡S及中国北京CH-BJ等GenBank报道的7个不同地域、不同种寄主植物上的分离物序列进行同源性分析的结果显示:ZYMV新疆分离物的CP基因核苷酸与其它11种ZYMV分离物的同源性高达99.42%,氨基酸序列同源性高达98.48%;新疆5个来源于不同寄主植物的ZYMV,不存在明显的遗传变异性,同源性高达97%,而与北京CH-BJ,Reunion和Singapore分离物相比存在较大差异,说明ZYMV地域的相关性要大于寄主上相关性。

Molecular Identification and Sequence Analysis for Zuccihini Yellow Masaic Virus Isolates in Xinjiang

A variety of Cucurbitaceae crops with yellow mosaic,yellow and blister spot were collected In Shihezi,Xinjiang.The coat protein gene of Zucchini yellow mosaic virus(ZYMV) was amplified via RT-PCR from total RNA of infect Cucumis sativus,Cucumis melo,and Cuffa cyliudrica leaves and cloned pMD18-T vector.Sequence analysis indicated that the nucleotide fragment containing 1185bp and includes complete CP gene.The obtained sequences were compared with other 7 reported ZYMV which were isolated from different regions and host plants.Nucleic acid sequence of CP gene was found relating to their host origin but little difference with geological distribution.Similar results were obtained from the CP amino acid sequences deduced from nucleic acid sequences.