西瓜花叶病毒(WMV)外壳蛋白基因克隆及序列分析  

Coat Protein Gene Cloning and Sequence Analysis of Watermelon Mosaic Virus (WMV) in Xinjiang Main Planting Areas

作　　者：何丹[1] 海利力.库尔班 玉山江.麦麦提 李继阳[1] 杨渡[2] 韩盛[2] 

机构地区：[1]新疆农业大学林学与园艺学院,乌鲁木齐830091 [2]新疆农业科学院植物保护研究所,乌鲁木齐830091

出　　处：《新疆农业科学》2015年第10期1849-1858,共10页Xinjiang Agricultural Sciences

基　　金：新疆农业科学院青年基金项目(xjnkq-2013033);中国科学院"西部之光"人才培养计划项目(YB201307)~~

摘　　要：【目的】研究新疆甜瓜主栽区西瓜花叶病毒(WMV)外壳蛋白的分类地位,以及与国内外分离株亲缘关系。【方法】田间采集具有西瓜花叶病毒症状的幼嫩叶片,利用改良的TRIZOL总RNA提取法获取RNA,进一步分离纯化、克隆及测序,将测序结果利用BLAST、DNAMAN 5.0、CLUSTALX、MEGA 5.0 EXPASY、TMHMM等软件进行碱基和氨基酸序列比对,以及疏水性平均值、脂肪系数、亲缘关系等分析。【结果】序列比对发现8个地区WMV病毒外壳蛋白核酸序列的5'端保守性较高,碱基和氨基酸序列相似性分别达到86.25%和93.01%,存在部分碱基和氨基酸突变,相似性较高。【结论】通过分子进化树可得出新疆8个分离物之间亲缘性关系较近,同源性较高,与国内外其它地区的亲缘性关系较远、同源性较低。[ Objective] To study the watermelon mosaic virus （WMV） taxonomic status in Xinjiang Main Planting Areas and kinship coat protein isolates at home and abroad. [ Method ] The young leaves taken from fields had watermelon mosaic virus symptoms, RNA was extracted using a modified method, and further separated, purified, cloned and sequenced. Analyses of the nucleotide and amino acid sequence alignment and hydrophobic average coefficient of fat EXPASY, TMHMM and other originals were carried out from the results of the sequencing by the means of the Internet, DNAMAN 5.0, CLUSTAL X, and MEGA 5.0. [ Result ] The sequence alignments found the 5 and of the eight regions WMV viral nucleic acid sequences conserved high nucleotide and amino acid sequence similarity reach 86.25% and 93.01%, higher similarity. The existence of partial nucleotide and amino acid mutations, the affinity between the eight isolates in Xinjiang had closer affinity relationships with other regions farther abroad.

关 键 词：甜瓜 西瓜花叶病毒 外壳蛋白 序列分析 

分 类 号：S436.5[农业科学—农业昆虫与害虫防治] S188[农业科学—植物保护]