板蓝根花叶病病原分子鉴定  

Molecular Identification of Pathogen of Isatis indigotica Mosaic

作　　者：魏若凡 王蓉[1] 李勇[1] 丁万隆[1] WEI Ruo-fan;WANG Rong;LI Yong;DING Wan-long(Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100193,China)

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193

出　　处：《中国现代中药》2021年第9期1601-1605,共5页Modern Chinese Medicine

基　　金：中央本级重大增减支项目(2060302);中国医学科学院医学与健康科技创新工程项目(2016-I2M-3-017)。

摘　　要：目的:检测安徽省太和县板蓝根花叶病病原。方法:采用反转录聚合酶链式反应(reverse transcription-polymerase chain reaction,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外壳蛋白(coat protein,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: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中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。结论:明确了安徽省太和县板蓝根花叶病的病原为CMV。Objective:To identify the pathogen(s)of mosaic in Isatis indigotica in Taihe County,Anhui Province.Methods:In this study,reverse transcription-polymerase chain reaction(RT-PCR)was performed on 5 I.indigotica mosaic leaf samples with specific primers for cucumber mosaic virus(CMV),broad bean wilt virus 2(BBWV2),and Potyvirus.The resultant products of two samples(AH-BLG3 and AH-BLG4)were sequenced,followed by BLAST analysis of pathogen species and molecular characteristics,and construction of the phylogenetic tree according to the amino acid sequence of coat protein(CP)of CMV.Thereby,the taxa of the pathogen were clarified.Results:All 5 samples were infected by CMV and BBWV2 and Potyvirus were not detected.The CP genes of isolates AH-BLG3 and AH-BLG4 shared the highest identity with CMV ZJAm which was isolated from Atractylodes macrocephala,with 99.54%and 98.93%%nucleotide identity,and 100%amino acid identity,respectively.Phylogenetic analysis showed that isolates AH-BLG3 and AH-BLG4 had the closest kinship with CMV ZJAm and Anhui and belonged to the subgroup IB.Conclusion:CMV was identified as the pathogen of I.indigotica mosaic in Anhui Province.

关 键 词：板蓝根 黄瓜花叶病毒 反转录聚合酶链式反应 外壳蛋白 系统进化分析 

分 类 号：R282.5[医药卫生—中药学]