板蓝根花叶病病原分子鉴定
投稿时间:2020-11-30  修订日期:2021-02-19   点此下载全文
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作者中文名作者英文名单位中文名单位英文名E-Mail
魏若凡 Wei Ruofan 中国医学科学院北京协和医学院药用植物研究所 Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &
Peking Union Medical College
1461104385@qq.com 
王蓉 Wang Rong 中国医学科学院北京协和医学院药用植物研究所 Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &
Peking Union Medical College
wangrong1019@163.com 
李勇 Li Yong 中国医学科学院北京协和医学院药用植物研究所 Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &
Peking Union Medical College
liyong@implad.ac.cn 
丁万隆* Ding Wanlong 中国医学科学院北京协和医学院药用植物研究所 Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &
Peking Union Medical College
wlding@implad.ac.cn 
基金项目:中国医学科学院医学与健康科技创新工程 (2016-I2M-3-017);中央本级重大增减支项目“名贵中药资源可持续利用能力建设”(2060302)
中文摘要:安徽省太和县发现板蓝根花叶病,该病害成为为害板蓝根生产的一种重要病害。为此,本研究采用RT-PCR方法,使用黄瓜花叶病毒(Cucumber mosaic virus,CMV)、蚕豆萎蔫病毒2号(Broad bean mosaic virus 2,BBWV2)和马铃薯Y病毒属(Potyvirus)引物,对5个板蓝根花叶样品进行分子鉴定。选取2个样品(AH-BLG3和AH-BLG4)的扩增产物进行测序,BLAST分析病原种类及分子特征,根据CMV CP氨基酸序列构建系统发育进化树明确病原的分类地位。结果显示:5个板蓝根样品均被CMV侵染,未检测到BBWV2和马铃薯Y病毒属病毒。BLASTn分析发现,该CMV板蓝根分离物与我国白术上的ZJAm分离物相似度最高,核苷酸相似度分别为 98.93%和99.54%,氨基酸相似度均为100%。系统进化分析发现该CMV板蓝根分离物与ZJAm和Anhui分离物亲缘关系最近,同属CMV IB亚组。
中文关键词:板蓝根  黄瓜花叶病毒  RT-PCR检测  外壳蛋白  系统进化分析
 
Molecular identification of the pathogen causing Isatis mosaic disease
Abstract:Isatidis mosaic disease was found in Taihe County, Anhui Province, and this disease has become an important disease threatening the production of Isatidis Radix. To identify the causal agent(s) of this disease, reverse transcription (RT)-PCR was performed with specific primers detecting Cucumber mosaic virus (CMV), Broad bean wilt virus 2 (BBWV2) and viruses in the genus Potyvirus. The results showed that all 5 samples were infected by CMV. No BBWV2 and potyvirus were detected in all of them. Isolates AH-BLG3 and AH-BLG4 were sequenced and used for BLASTn analysis and phylogenetic analysis. The results indicated that these two fragments shared the highest nucleotide identity with CMV ZJAm which isolated from Atractylodes macrocephala, with 99.54% and 98.93%% nucleotide identity respectively, and 100% amino acid identity. Phylogenetic analysis based on amino acid sequences of CMV CPs showed that isolate AH-BLG3 and AH-BLG4 clustered with CMV ZJAm and Anhui, and belongs to the subgroup IB.
keywords:Isatis indigotica  Cucumber mosaic virus  RT-PCR detection  Coat protein  Phylogenetic analysis
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