收稿日期: 2021-07-09
录用日期: 2021-11-20
网络出版日期: 2021-11-17
基金资助
宁夏回族自治区农业特色优势产业育种专项(NXNYYZ20200101);宁夏回族自治区重点研发计划(2021BBF02002);宁夏回族自治区重点研发计划(重大)重点项目(2019BBF02022)
Molecular Identification of Tomato Spotted Wilt Virus on Tomato in Yinchuan
Received date: 2021-07-09
Accepted date: 2021-11-20
Online published: 2021-11-17
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394 bp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。
关键词： 银川; 番茄斑萎病毒; RT-PCR; Western blot
王晓敏, 李洪磊, 王林, 周鹏泽, 白圣懿, 李国花, 郑福顺, 陶小荣, 程国新, 高艳明, 李建设 . 银川番茄斑萎病毒的分子鉴定[J]. 植物学报, 2021 , 56(6) : 715 -721 . DOI: 10.11983/CBB21113
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.
Key words： Yinchuan; tomato spotted wilt virus; RT-PCR; Western blot
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