[1]干射香,涂丽琴,吴淑华,等.山东寿光黄瓜上瓜类褪绿黄化病毒的分子鉴定[J].江苏农业学报,2019,(05):1047-1053.[doi:doi:10.3969/j.issn.1000-4440.2019.05.007]
　GAN She-xiang,TU Li-qin,WU Shu-hua,et al.Molecular identification of cucurbit chlorotic yellows virus on cucumber in Shouguang, Shandong province[J].,2019,(05):1047-1053.[doi:doi:10.3969/j.issn.1000-4440.2019.05.007]
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山东寿光黄瓜上瓜类褪绿黄化病毒的分子鉴定()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2019年05期
页码:
1047-1053
栏目:
植物保护
出版日期:
2019-10-31

文章信息/Info

Title:
Molecular identification of cucurbit chlorotic yellows virus on cucumber in Shouguang, Shandong province
作者:
干射香1; 2; 涂丽琴2; 吴淑华2; 范小燕2; 吴瑞雪1; 2; 赵文浩2; 程兆榜2; 周益军2; 竺晓平3; 季英华2; 章松柏1
(1.长江大学农学院，湖北荆州434025；2.江苏省农业科学院植物保护研究所/省部共建国家重点实验室培养基地——江苏省食品质量安全重点实验室，江苏南京210014；3.山东农业大学植物保护学院，山东泰安271018)
Author(s):
GAN She-xiang1; 2; TU Li-qin2; WU Shu-hua2; FAN Xiao-yan2; WU Rui-xue1; 2; ZHAO Wen-hao2; CHENG Zhao-bang2; ZHOU Yi-jun2; ZHU Xiao-ping3; JI Ying-hua2; ZHANG Song-bai1
(1.College of Agriculture, Yangtze University, Jingzhou 434025, China;2.Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences State Key Laboratory Breeding Base-Key Lab of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China;3.College of Plant Protection, Shandong Agricultural University, Taian 271018, China)
关键词:
黄瓜; 瓜类褪绿黄化病毒; 分子鉴定
Keywords:
cucumber; cucurbit chlorotic yellows virus; molecular identification
分类号:
S642.2
DOI:
doi:10.3969/j.issn.1000-4440.2019.05.007
文献标志码:
A
摘要:
2016年，本研究在对山东黄瓜病毒病调查时发现当地黄瓜上出现一种叶片表现黄化植株伴有轻微矮缩的病害，为明确其中伴随的病毒种类，我们对田间病样进行了采集，提取总RNA后利用瓜类病毒引物进行RT-PCR检测，结果发现在利用瓜类褪绿黄化病毒( Cucurbit chlorotic yellows virus，CCYV)引物检测样品时，田间采集的25份疑似病样中13份能扩增到400 bp左右的目的条带。为进一步确认，我们又针对CCYV的外壳蛋白基因(CP)设计了特异性引物，对13份阳性样品进行检测，结果均能扩增到目的片段，对获得的CP基因片段进行克隆并测定序列，序列分析结果显示其CP基因序列全长753 bp，编码1个由250个氨基酸组成的分子量28 700的蛋白质。进一步序列分析结果显示其与CCYV已报道分离物有很高的同源性，并聚类到同一个大的分支，其中与日本分离物、希腊分离物及中国其他地区分离物相对近缘，聚类到一支，而与伊朗分离物近缘关系相对较远。这些结果说明山东黄瓜黄化病样上检测到的病毒为瓜类褪绿黄化病毒，这也是该病毒在山东侵染黄瓜的首次报道。
Abstract:
An investigation on cucumber virus in Shandong province was carried out in 2006 and a cucumber disease with yellowing leaves and mild dwarf symptoms was found. To charify the types of viruses associated with the disease, 25 samples were collected and RNA was extracted. RT-PCR was performed using diverse cucumber-infecting virus primers and a fragment about 400 bp was amplified from 13 samples using primers CCYV-HSPS1 and CCYV-HSPS2, indicating that cucurbit chlorotic yellows virus (CCYV) was present in these samples. To further confirm the result, a pair of specific primers CCCP-F and CCCP-R were designed based on the coat protein(CP) gene of CCYV, and the target fragment was amplified from all 13 samples. Sequence analysis results showed that the CP obtained in this study was 735 bp in length and encoded a protein consisting of 250 amino acids with molecular weight of 28 700. Further sequence analysis results showed that it had high homology with other CCYV isolates, including Japanese isolates, Greece isolates and other isolates from China. All these results indicate that the virus detected in cucumber from Shandong province is CCYV, this is the first report of CCYV infecting cucumber in Shandong province.

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备注/Memo

备注/Memo:
收稿日期：2018-11-28 基金项目：国家重点研发计划项目(2018YFD0201208)；国家特色蔬菜产业技术体系项目(CARS-24-C-01)；国家自然科学基金项目(31572074、31770168)；省部共建国家重点实验室培育基地自主研究课题(zb201705)；江苏省农业科技自主创新基金项目(CX(18)2005)；江苏省农业科学院基金项目(6111614) 作者简介：干射香(1994-)，女，湖北黄冈人，硕士，研究方向为分子植物病毒学。 通讯作者：章松柏,(E-mail)yangtze2008@126.com；季英华,(E-mail)jiyinghua@jaas.ac.cn
更新日期/Last Update: 2019-11-11