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园艺学报 ›› 2020, Vol. 47 ›› Issue (7): 1391-1400.doi: 10.16420/j.issn.0513-353x.2019-0920

• 研究报告 • 上一篇    下一篇

青海辣椒上番茄斑萎病毒检测及鉴定

吴淑华1,*,涂丽琴1,*,咸文荣2,闫佳会2,高丹娜1,吉 颖2,陶小荣3,周益军1,郭青云2,**,季英华1,**   

  1. 1江苏省农业科学院植物保护研究所,南京 210014;2青海大学农林科学院,青海省农业有害生物综合治理重点实验室,农业部西宁作物有害生物科学观测实验站,西宁 810016;3南京农业大学植物保护学院,南京 210095
  • 出版日期:2020-07-25 发布日期:2020-07-25
  • 基金资助:
    国家重点研发计划项目(2018YFD0201208);国家自然科学基金项目(31572074,31770168);国家现代农业产业技术体系建设专项资金项目(CARS-24-C-01);江苏省农业自主创新基金项目[CX(18)2005];江苏省农业科学院基金项目(6111614);青海省科技成果转化专项(2018-NK-127)

Detection and Identification of Tomato spotted wilt tospovirus on Pepper from Qinghai Province

WU Shuhua1,*,TU Liqin1,*,XIAN Wenrong2,YAN Jiahui2,GAO Danna1,JI Ying2,TAO Xiaorong3,ZHOU Yijun1,GUO Qingyun2,**,and JI Yinghua1,**   

  1. 1Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;2Academy of Agriculture and Forestry Sciences,Qinghai University,Key Laboratory of Agricultural Integrated Pest Management, Scientific Observing and Experimental Station of Crop Pests in Xining,Ministry of Agriculture,Xining 810016,China; 3College of plant protection,Nanjing Agricultural University,Nanjing 210095,China
  • Online:2020-07-25 Published:2020-07-25

摘要: 在对青海辣椒上的病毒病进行调查时发现疑似番茄斑萎病毒(Tomato spotted wilt tospovirus,TSWV)病样,表现为环斑、坏死等症状。为明确其感染的病毒种类,对其进行了检测和鉴定。对Tospovirus的通用引物进行检测时发现采集的49份样品中有14份样品扩增到了目的条带,序列测定结果显示其与TSWV的同源性最高;同时利用TSWV特异性引物对阳性样品进行检测,结果全部为阳性;基于克隆的NSs基因序列聚类分析结果显示其属于TSWV的1个分离物,与中国云南、黑龙江等地分离物相对近缘,而与国外其他分离物相对远缘。在分子鉴定的基础上,利用TSWV的抗体通过Western blot进一步对样品进行了检测,结果也证明采集样品中存在TSWV的感染。这些结果表明青海辣椒上存在TSWV的感染。

关键词: 辣椒, 番茄斑萎病毒, 青海, 检测

Abstract: An investigation on pepper virus in Qinghai province was carried out and samples with leaf spot and necrosis symptoms was found in the fields,which was very similar with the symptoms induced by Tomato spotted wilt tospovirus(TSWV). To identify the possible virus associated with the disease,49 samples was collected and detected with various pepper viruses using RT-PCR and 14 samples was found positive when using a pair of degenerate primers for Tospovirus. Samples were selected randomly for sequencing and the result of sequence analysis revealed that the virus shared the highest sequence identities with TSWV. Then 14 samples were further detected using specific primers for TSWV and the results showed that all samples were detected. Further phylogenetic analysis based on the sequence of NSs also revealed that the virus detected was clustered with TSWV and shared the highest sequence identities with the isolates from China,including Yunnan and Heilongjiang and relatively lower identities with others abroad,which show that the virus is an isolate of TSWV. Further results based on western blot using TSWV antibody also confirmed that the samples were infected by TSWV. All these results showed that the virus infected pepper is an isolate of TSWV.

Key words: pepper, Tomato spotted wilt tospovirus, Qinghai, detection

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