基于DNA序列分析的葡萄茎枯病菌检疫鉴定  

Detection and identification of Didymella glomerata based on DNA sequence analysis

作　　者：段丽君 吕燕 张慧丽 段维军 Duan Lijun;Lu Yan;Zhang Huili;Duan Weijun(Ningbo Academy of Inspection and Quarantine,Ningbo 315012,China;Ningbo Joysun Product Testing Service Company;Technical Service Station of Animal Husbandry and Veterinary Medicine in Haishu District,Ningbo;Ningbo Customs District)

机构地区：[1]宁波检验检疫科学技术研究院,浙江宁波315012 [2]宁波中盛产品检测有限公司 [3]宁波市海曙区畜牧兽医技术管理服务站 [4]宁波海关

出　　处：《植物检疫》2020年第4期22-30,共9页Plant Quarantine

基　　金：国家重点研发计划(2016YFF0203201);原国家质检总局科研项目(2017IK059);中科院战略生物资源服务网络计划(KFJBRP-009);浙江省公益技术研究项目(LGF20C140001);宁波市自然科学基金(2019A610411)。

摘　　要：葡萄茎枯病菌是我国进境植物检疫性有害生物,目前口岸对葡萄茎枯病菌的检疫鉴定主要依靠形态学观察,然而单纯利用形态学难以对其进行准确鉴定,需与分子生物学方法相结合。为找出有效分子标记,本研究以宁波口岸截获鉴定的41株葡萄茎枯病菌分离株和收集自ATCC和CBS的2株葡萄茎枯病菌为研究对象,对上述菌株进行形态学观察,ITS、actin和β-tubulin基因序列测定,同时采用序列分析手段考察上述基因片段在葡萄茎枯病菌鉴定区分上的应用价值。研究结果表明葡萄茎枯病菌形态变异较大,ITS无法有效区分葡萄茎枯病菌,actin和β-tubulin基因片段能够将葡萄茎枯病菌从近似种中准确划分出来,利用3个基因片段联合建树区分能力较强。Abstact Didymella glomerata is listed as plant quarantine pest of China. The current quarantine identification method for D. glomerata is mainly based on morphological method. However,D. glomerate is difficult to be identified accurately only based on morphology,and integration of molecular and morphological data are further needed. To find effective molecular markers,43 isolates of the D. glomerata,collected from Ningbo Port,ATCC and CBS,were cultivated for morphological observation and studied by ITS,actin and β-tubulin sequences analyses to differentiate the D. glomerata from others. Analysis of these sequences with the related sequences in NCBI show that ITS can’t effectively distinguish between D. glomerata and others,however,actin and β-tubulin fragments can accurately distinguish it. This study also suggests that combination analysis of the ITS,actin and β-tubulin sequences is an accurate method to differentiate the D. glomerata from other species.

关 键 词：葡萄茎枯病菌 检疫性真菌 系统发育 序列分析 鉴定 

分 类 号：S41-30[农业科学—植物保护]