芸薹根肿菌生理小种鉴定和分子标记  

Identification of pathotype and development of molecular marker for Plasmodiophora brassicae causing clubroot

作　　者：王神云[1,2] 向明艳 王红[2] 于利[2] 李建斌[2] 江解增[1] WANG Shenyun XIANG Mingyan WANG Hong YU Li LI Jianbin Jiang Jizeng(Coil of Hort and Plant Prot, Yangzhou Univ, Yangzhou 225000, China Inst of Veg Crops, Jiangsu Acad of Agr Sci, Nanjing 210014, China Agri Serv Cent of Wangying Coun, Lichuan City, Lichuan, 445400, China)

机构地区：[1]扬州大学园艺与植物保护学院,江苏扬州225009 [2]江苏省农业科学院蔬菜研究所,江苏南京210014 [3]利川市汪营镇农业服务中心,湖北利川445400

出　　处：《扬州大学学报（农业与生命科学版）》2016年第4期91-96,共6页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家现代农业产业技术体系项目(CARS-25);江苏省科技支撑计划项目(BE2013429);江苏省农业科技自主创新基金项目[CX(13)2004]

摘　　要：采集湖北利川结球甘蓝病发生严重区域的病土和病根,通过形态和PCR技术鉴定,发现病原菌的形态特征与芸薹根肿菌(Plasmodiophora brassicae Wor.)相似,芸薹根肿菌检测引物从被感染根组织中扩增到特异带,明确为芸薹根肿菌。随机选2条特异带进行序列分析,均与P.brassicae ITS保守区域序列高度同源,一致性达到99%以上,而且Plasmo3-4扩增特异带序列囊括在PbITS1-2扩增的特异带中。随后,Plasmo3-4引物在10个区域根肿菌和19个十字花科代表作物进行了检测,所有根肿菌扩增出近300bp特异带,所有作物中没有任何条带,表明Plasmo3-4特异引物可以直接快速地用于根肿菌检测;利川病原菌检测后,进一步利用欧洲ECD鉴别系统鉴定此芸薹根肿菌生理小种为ECD16/4/0,致病力一般。芸薹根肿菌及其致病性鉴定为抗病研究和抗病育种奠定了基础。In the present study,infested soil samples and clubbed roots were collected for identification of the causal agent in clubroot-affected area in Lichuan,Hubei Province.The identification by morphology and PCR technology revealed that the pathogen shared similar morphology with Plasmodiophora brassicae.PCR specific amplification from clubbed roots suggested that the pathogen was P.brassicae.Sequencing randomly of two fragments amplified with primer pairs PbITS1-2and Plasmo3-4showed that both fragments shared high similarity with internal transcribed spacer（ITS）of P.brassicae.It was interesting that the band of Plasmo3-4was among PbITS1-2band,and molecular detection of 19 representative crops of crucifers and ten P.brassicae strains with primer Plasmo3-4revealed that Plasmo3-4was specific for rapid detection of all P.brassicae strains,but not healthy crucifer crops.Furthermore,the pathotype of P.brassicae pathogen was identified as ECD16/4/0according to ECD detection system.The detection of pathogen and pathotype would be helpful for resistance research and breeding.

关 键 词：根种病 芸薹根肿菌 分子标记 生理小种 

分 类 号：S635.1[农业科学—蔬菜学] S432.42[农业科学—园艺学]