银川番茄斑萎病毒的分子鉴定  

Molecular Identification of Tomato Spotted Wilt Virus on Tomato in Yinchuan

作　　者：王晓敏[1,2,3,4] 李洪磊 王林[1] 周鹏泽 白圣懿[1] 李国花 郑福顺 陶小荣 程国新 高艳明[1,2,3,4] 李建设 Xiaomin Wang;Honglei Li;Lin Wang;Pengze Zhou;Shengyi Bai;Guohua Li;Fushun Zheng;Xiaorong Tao;Guoxin Cheng;Yanming Gao;Jianshe Li(School of Agriculture,Ningxia University,Yinchuan 750021,China;Ningxia Modern Facility Horticulture Engineering Technology Research Center,Yinchuan 750021,China;Key Laboratory of Modern Molecular Breeding for Dominant and Special Crops in Ningxia,Yinchuan 750021,China;Ningxia Facility Horticulture(Ningxia University)Technology Innovation Center,Yinchuan 750021,China;College of Plant Protection,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]宁夏大学农学院,银川750021 [2]宁夏现代设施园艺工程技术研究中心,银川750021 [3]宁夏优势特色作物现代分子育种重点实验室,银川750021 [4]宁夏设施园艺(宁夏大学)技术创新中心,银川750021 [5]南京农业大学植物保护学院,南京210095

出　　处：《植物学报》2021年第6期715-721,共7页Chinese Bulletin of Botany

基　　金：宁夏回族自治区农业特色优势产业育种专项(No.NXNYYZ20200101);宁夏回族自治区重点研发计划(No.2021BBF02002)和宁夏回族自治区重点研发计划(重大)重点项目(No.2019BBF02022)。

摘　　要：为明确银川番茄(Lycopersicon esculentum)是否遭受了番茄斑萎病毒(TSWV)的危害,采用国家标准TSWV RT-PCR检测技术对银川番茄上采集的14份疑似感染TSWV病叶样本进行分子鉴定,对克隆得到的核衣壳蛋白基因N(Nucleocapsid)序列进行多序列比对和系统进化树分析,随后对PCR阳性样本进行蛋白检测。结果表明,14份病叶样本中有8份扩增出长度为394bp的TSWV N基因序列,且8条序列完全一致;获得的银川番茄TSWV分离物与云南番茄、中国莴苣(Lactuca sativa)、中国鸢尾(Iris tectorum)和重庆辣椒(Capsicum annuum)TSWV分离物相对近缘,与山东、黑龙江和北京等地及国外TSWV分离物相对远缘;利用TSWV的抗体通过Western blot对8个PCR阳性样本进一步检测,结果也证实8个阳性样本中存在TSWV感染。该研究首次通过分子鉴定及蛋白检测证明银川番茄上存在TSWV感染,需要加快抗TSWV番茄品种的选育工作。In order to determine whether Yinchuan tomato(Lycopersicon esculentum)was damaged by tomato spotted wilt virus(TSWV),14 disease leaf samples of suspected infected TSWV collected from Yinchuan tomato were identified by the national standard TSWV RT-PCR technique.The cloned N gene sequences were analyzed by multi sequence alignment and phylogenetic tree analysis.Eight of the 14 disease leaf samples amplified the TSWV N gene sequence with a length of 394 bp,and the 8 sequences were completely consistent.The TSWV sequence was relatively close to those from Yunnan tomato,Chinese lettuce(Lactuca sativa),Chinese Iris(Iris tectorum)and Chongqing pepper(Capsicum annuum),while relatively distant to those from Shandong,Heilongjiang,Beijing and the abroad.Detection of TSWV in the 8 PCR positive samples by use of Western bolt showed that TSWV infection existed in these samples.This study proved that there was TSWV infection in Yinchuan tomato.Therefore,it is necessary to speed up the breeding of TSWV resistant tomato varieties.

关 键 词：银川 番茄斑萎病毒 RT-PCR Western blot 

分 类 号：S436.412.11[农业科学—农业昆虫与害虫防治]