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瓜类褪绿黄化病毒山东分离物全基因组序列扩增及分析

引用本文：臧连毅,孙晓辉,苏文敏,张先平,刘晓莹,竺晓平.瓜类褪绿黄化病毒山东分离物全基因组序列扩增及分析.植物保护学报,2019,46(6):1195-1202

DOI：10.13802/j.cnki.zwbhxb.2019.2018181

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作者	单位	E-mail
臧连毅	山东农业大学植物保护学院, 山东省蔬菜病虫生物学重点实验室, 泰安 271018
孙晓辉	山东农业大学植物保护学院, 山东省蔬菜病虫生物学重点实验室, 泰安 271018
苏文敏	上海市浦东新区高东镇城镇建设管理中心, 上海 200120
张先平	山东农业大学植物保护学院, 山东省蔬菜病虫生物学重点实验室, 泰安 271018
刘晓莹	山东农业大学植物保护学院, 山东省蔬菜病虫生物学重点实验室, 泰安 271018
竺晓平	山东农业大学植物保护学院, 山东省蔬菜病虫生物学重点实验室, 泰安 271018	zhuxp@sdau.edu.cn

中文摘要:为明确分离自山东省寿光市甜瓜上的瓜类褪绿黄化病毒（cucurbit chlorotic yellows virus，CCYV）分离物的全基因组序列信息和遗传变异情况，利用毛形病毒属Crinivirus简并引物进行RTPCR检测，利用RACE技术结合RT-PCR方法克隆CCYV山东分离物2条RNA链的全基因组序列，通过与GenBank中其它地区CCYV分离物的全长序列进行比对分析其同源性，并基于CP基因序列构建系统进化树分析其遗传变异情况。结果表明，山东分离物经RT-PCR检测和测序后确定为CCYV。CCYV山东分离物与其它CCYV分离物的RNA1链和RNA2链的全基因组序列一致性的平均值分别为99.82%和99.88%，且2条链的5'末端均比较保守，没有碱基突变的情况发生；RNA1链3'末端存在2个碱基变异，RNA2链3'末端存在1个碱基变异。CCYV不同地区分离物主要分为3个簇群，其中山东分离物和中国其它地区分离物、日本分离物、苏丹分离物、黎巴嫩分离物和塞浦路斯分离物聚类在一起。研究表明CCYV基因组序列比较保守，该病毒的分化可能与地理来源存在一定的相关性。

中文关键词:瓜类褪绿黄化病毒全基因组系统进化树

Amplification and analysis of whole genome sequence of cucurbit chlorotic yellows virus from Shandong Province, China

Author Name	Affiliation	E-mail
Zang Lianyi	Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China
Sun Xiaohui	Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China
Su Wenmin	Gaodong Urban Construction Management Center, Pudong New Area, Shanghai 200120, China
Zhang Xianping	Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China
Liu Xiaoying	Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China
Zhu Xiaoping	Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests, College of Plant Protection, Shandong Agricultural University, Tai'an 271018, Shandong Province, China	zhuxp@sdau.edu.cn

Abstract:To clarify the whole-genome sequence and genetic variation of cucurbit chlorotic yellows virus (CCYV) isolated from melon in Shouguang City, Shandong Province, degenerate primers of the genus Crinivirus were used for RT-PCR detection, and the whole-genome sequences of two RNA strands of CCYV Shandong isolate were obtained by RACE and RT-PCR cloning. The similarity of the wholegenome sequence between Shandong isolate and isolates of CCYV from other regions in GenBank was compared. The genetic variation was analyzed by constructing the phylogenetic tree based on the coat protein (CP) gene sequences. The results showed that the isolate from Shandong was identified as CCYV by RT-PCR detection and sequencing, and the average sequence identity of RNA1 and RNA2 among Shandong isolate and other isolates were 99.82% and 99.88%, respectively. The 5' ends of the two strands of Shandong isolate were both conservative and no base mutation occurred. Furthermore, there were two base variations at the 3' end of the RNA1 strand, and one base variation at the 3' end of the RNA2 strand. The isolates from different geographical origins of CCYV were mainly divided into three clusters. The isolate from Shandong was clustered with isolates from other areas of China, Japanese isolates, Sudan isolate, Lebanese isolates and Cyprus isolates. This study indicated that the genomic sequence of CCYV was relatively conserved and that differentiation of the virus might be related to geographical occurrence.

keywords:cucurbit chlorotic yellows virus whole-genome sequences phylogenetic tree

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