[1]涂丽琴,吴淑华,干射香,等.江苏省蚕豆上菜豆黄花叶病毒的分子鉴定[J].江苏农业学报,2019,(04):804-810.[doi:doi:10.3969/j.issn.1000-4440.2019.04.008]
　TU Li qin,WU Shu hua,GAN She xiang,et al.Molecular identification of bean yellow mosaic virus infecting Vicia faba from Jiangsu province[J].,2019,(04):804-810.[doi:doi:10.3969/j.issn.1000-4440.2019.04.008]
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江苏省蚕豆上菜豆黄花叶病毒的分子鉴定()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2019年04期
页码:
804-810
栏目:
植物保护
出版日期:
2019-08-31

文章信息/Info

Title:
Molecular identification of bean yellow mosaic virus infecting Vicia faba from Jiangsu province
作者:
涂丽琴1; 2; 吴淑华2; 干射香2; 崔晓艳3; 赵文浩2; 程兆榜2; 陈新3; 周益军2; 季英华2; 朱月林1
（1.南京农业大学园艺学院，江苏南京210095；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.江苏省农业科学院经济作物研究所，江苏南京210014）
Author(s):
TU Liqin1; 2; WU Shuhua2; GAN Shexiang2; CUI Xiaoyan3; ZHAO Wenhao2; CHENG Zhaobang2; CHEN Xin3; ZHOU Yijun2; JI Yinghua2; ZHU Yuelin1
（1. College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China;2.Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences/Jiangsu Key Laboratory for Food Quality and SafetyState Key Laboratory Cultivation Base of Ministry of Science and Technology， Nanjing 210014, China;3.Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences， Nanjing 210014, China）
关键词:
菜豆黄花叶病毒; 蚕豆; 分子鉴定
Keywords:
bean yellow mosaic virus; Vicia faba; molecular identification
分类号:
S643
DOI:
doi:10.3969/j.issn.1000-4440.2019.04.008
文献标志码:
A
摘要:
为掌握江苏省豆类作物上的病毒病种类，本研究于2018年对江苏省蚕豆作物病毒进行了调查，累计采集田间疑似病样55份，为明确其中伴随的病毒种类，我们对田间采集的疑似病样提取总RNA后进行了分子检测，结果发现，在用菜豆黄花叶病毒（Bean yellow mosaic virus，BYMV）检测引物进行RTPCR扩增时有16份样品扩增到1条525 bp左右的条带，与目的片段大小一致，说明其中可能伴随有菜豆黄花叶病毒的感染。为进一步确定这一结果，我们针对BYMV CP基因设计了引物，以阳性样品RNA为模板进行RTPCR，结果发现均扩增到预期大小的目的条带，对该片段回收、克隆并测定碱基序列，结果发现其CP基因碱基序列全长819 bp，编码1个由273个氨基酸组成大小约20 800的蛋白质。BLAST分析结果显示其与BYMV同源性最高（AB439731，99%），表明其属于BYMV的一个分离物。利用MEGA软件对其进行系统进化分析，结果发现这些分离物与日本的蚕豆分离物（AB439731）同源性最高，其次是澳大利亚的蚕豆分离物（HG970867）和中国青海的菜豆分离物（916）。这些结果表明江苏蚕豆上检测到的病毒是菜豆黄花叶病毒的一个分离物，这是该病毒在江苏侵染蚕豆的首次分子鉴定报道。
Abstract:
In order to have a better understanding about the virus infecting bean in Jiangsu province, an investigation on faba bean virus was carried out and 55 diseased samples were collected in 2018. Total RNA was extracted and RTPCR was performed to clarify the virus associated with the diseased samples and a fragment about 525 bp was amplified from 16 samples using primers BYMVF1/BYMVR2, indicating bean yellow mosaic virus (BYMV) was present in these samples. To further confirm the result, specific primers were designed based on coat protein of BYMV and the target fragment was amplified from all 16 samples. Three samples were selected randomly for cloning. The sequencing and sequence analysis results showed that the CP obtained in this study was 819 bp in length and encoded a protein consisting of 273 amino acids with a molecular weight of approximately 20 800. BLAST results showed that it had the highest identity with BYMV (AB439731, 99%) and sequence phylogenetic analyses results also showed that they shared the highest sequence identity with Japanese isolates of BYMV (AB439731, BYMV902), and clustered with Australian isolates (HG970867, BYMVFB) and Qinghai isolates (916) in the branch of BYMV. All these results show that the virus detected in faba bean from Jiangsu province is an isolate of BYMV, this is the first molecular identification report of BYMV infecting faba bean in Jiangsu province.

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备注/Memo

备注/Memo:
收稿日期：2018-10-19 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）；国家食用豆产业技术体系项目( CARS-08G15)；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]；江苏现代农业（特粮特经）产业技术体系集成创新中心项目[SXGC(2017)260]；江苏省农业科学院基金项目（6111614） 作者简介：涂丽琴（1996-），女，江西吉安人，硕士研究生，研究方向为园艺蔬菜作物病毒。吴淑华为共同第一作者。 通讯作者：朱月林，（E-mail）ylzhu@njau.edu.cn；季英华，（E-mail）jiyinghua@jaas.ac.cn
更新日期/Last Update: 2019-08-31