小西葫芦黄花叶病毒石河子分离物的分子变异  

Molecular Variability Analysis of Zucchini Yellow Mosaic Virus Isolates from Shihezi

作　　者：王志江[1] 杨海燕[1] 向本春[1] 郑银英[1] 

机构地区：[1]石河子大学生命科学学院/石河子大学农业生物技术重点实验室,石河子832003

出　　处：《石河子大学学报（自然科学版）》2015年第6期677-682,共6页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金项目(31460466);国际科技合作与交流专项(20072072)

摘　　要：为了研究小西葫芦黄花叶病毒(Zucchini yellow mosaic virus,ZYMV)石河子分离物的分子变异情况,本研究采用ELISA及一步法RT-PCR对2010-2014年采自石河子地区的30个葫芦科植物样品进行ZYMV检测,从阳性样品中通过一步法RT-PCR克隆其ZYMV分离物的CP基因,经PCR-SSCP分析,将带型一致的ZYMV分离物进行归类,每种类型中选取1个代表性的分离物进行CP基因测序,分析其核苷酸及推导的氨基酸序列的差异。结果显示:2种检测方法的检出率分别为63.33%和83.33%;根据ZYMV CP基因SSCP带型,将21个分离物分为5种不同的类型;其5个代表性分离物的CP核苷酸序列相似性为93.0%-99.8%,在系统进化树中位于2个不同的亚组;氨基酸序列的相似性为96.1%-99.6%,变异主要集中在N端的前41个氨基酸。这表明ZYMV石河子分离物间存在一定程度的分子变异。In order to learn the molecular variability of Zucchini yellow mosaic virus （ZYMV） isolates from Shihezi,in this study, ELISA and RT-PCR were used to detect ZYMV of 30 Cucurbitaceae crops from Shihezi region between 2010 and 2014.CP gene of ZYMV isolates was cloned from the positive samples by one-step RT-PCR.PCR-SSCP analysis were performed,and ZYMV isolates were classified depending on the consistent type of banding pattern.Representative isolates were chosen from each type for CP gene sequencing to analyze the nucleotide and deduced amino acid sequences.The results showed that the detected rates of two detection methods were 63.33% and 83.33%,respectively.21 isolates were divided into 5 different types depending on the SSCP banding pattern of ZYMV CP gene.The homology of CP nueleotide sequences of 5 representative isolates was 93.0%-99.8% and phylogenetic analysis indicated that they were grouped into 2 different subgroup.While the homology of CP amino acid sequences was 96.1%-99.6% and the variability mainly concentrated in 41 amino acids of N extremity.It concluded that molecular variability was existent up to a certain extent among ZYMV isolates from Shihezi.

关 键 词：小西葫芦黄花叶病毒 DAS—ELISA PCR—SSCP 分子变异 

分 类 号：S432.412[农业科学—植物病理学]