侵染新疆甜瓜的ZYMV衣壳蛋白基因克隆及其序列分析  

Cloning and Sequence Analysis for Coat Protein Gene of ZYMV Infecting of Melon(Cucumis melo) in Xinjiang

作　　者：李继洋[1] 杨渡[2] 韩盛[2] 何丹[1] 玉山江·麦麦提[2] 

机构地区：[1]新疆农业大学农学院,乌鲁木齐830000 [2]新疆农业科学院植物保护研究所/农业部西北荒漠绿洲作物有害生物综合治理重点实验室,乌鲁木齐830091

出　　处：《农业生物技术学报》2016年第5期718-728,共11页Journal of Agricultural Biotechnology

基　　金：中国科学院“西部之光”人才培养计划项目(No.YB201307)

摘　　要：新疆作为中国有名的瓜果之乡，其瓜类不仅成为其中最具代表性的品牌也日益成为农民增收致富的经济支柱产业。然而近年来由于新疆瓜果日益受到病毒病威胁，严重时造成大面积减产甚至绝收，大幅度制约了农民生产积极性和生活水平的提高。小西葫芦黄花叶病毒（Zucchini yellow mosaic virus，ZYMV）是其中比较具有代表性的病毒，为了对新疆不同主栽地区甜瓜（Cucumismelo）ZYMV进行生物防治和抗病毒甜瓜的研究。本研究利用同源克隆的方法，扩增获得了新疆19个样点ZYMV分离物衣壳蛋白（coatprotein．CP）基因全长序列，ZYMV分离物CP基因全长介于1080～1180bp，编码253～301个氨基酸，蛋白质大小约33kD。序列比对发现，ZYMV核酸序列的3’和5’非编码区末端保守性较低，编码区及其周围序列保守性较高，氨基酸序列的N端保守性高于c端。ZYMV同源性比对结果发现，病毒属于同一ZYMV基因型。通过核酸序列构建的分子进化树可以将ZYMV分为4个亚组，亚组分离依据符合地区分布特性。根据对蛋白质理化性质，包括蛋白质的动力学、化学和热力学稳定性分析得出，ZYMV均属于稳定性蛋白。结合新疆夏季平均降水量和平均气温变化趋势发现，其与对应地区ZYMVCP的疏水性指数和脂溶性指数的变化趋势存在一定相关性。分析认为，不同地区ZYMVCP的稳定性存在一定的差异，这可能与新疆特殊的气候条件与分离物所处地理环境有关。本研究通过序列分析获得19个样点ZYMVCP核酸序列以及CP特征，为今后以RNA干扰技术或基因组编辑技术培育抗病毒甜瓜，进行生物农药研发等提供了基础的数据支持，同时为开展对植物病毒的致病机理以及探究环境与病毒传播机制的研究提供初期的研究方向。Xinjiang is famous for fruit in China. And the melon is not only one of the most iconic brand, but the economic pillar industries that increase the farmers＇ income of home at recent years. However, due to the Xinjiang melon have been threatened increasingly by virus disease, causing a large area yield reduction even total loss of the melon yield when serious. It is affected seriously the farmers＇ production enthusiasm and the improvement of standard of living. And Zucchini yellow mosaic virus （ZYMV） is one of the most representative virual. In order to research biological control and anti-viral melon （Cucumis melo） in different cultivars region of Xinjiang, homologous cloning method were used, the coat protein （CP） complete gene sequence of ZYMV from 19 different regions of Xinjiang were amplified. The CP complete gene sequences of ZYMV were 1 080-1 180 bp, encoding 253-301 amino acids and synthetic protein size was approximately 33 kD. The result indicated that the nucleic acid sequence of 3＇ and 5＇ terminal noncoding region were less conserved, but the coding region and the surrounding sequence were high conserved. The N terminal of the amino acids was more conserved than C terminal. According to ZYMV homology results, isolator from 19 regions belonged to the same genotype of ZYMV. Through the analysis of the evolutionary tree that constructed according to nucleic acid sequences, ZYMV were divided into 4 subgroups, and the basis of subgroups were divided compliance with viral distribution character. The physical and chemical properties of proteins, such as the kinetics of protein, chemical and thermodynamic stability results showed that CP of ZYMV belonged to stable protein. The combined analysis with average summer precipitation and mean temperature trending in Xinjiang results showed that the change trends between index of hydrophobic and fat in CP of ZYMV with the corresponding region had a certain correlation. The analysis results indicated that the stability CP of ZYMV had some differen

关 键 词：新疆 小西葫芦黄瓜花叶病毒(ZYMV) 克隆 序列分析 蛋白稳定性 环境影响 

分 类 号：Q781[生物学—分子生物学]