福州北峰烟草青枯病强致病性病原菌的分离与鉴定  

Isolation and identification of strongly pathogenic pathogens of tobacco bacterial wilt in Beifeng,Fuzhou

作　　者：赵振富 夏莹子 林珅 刘新聪 程崖芝 余文 高文霞 巫升鑫 ZHAO Zhen-fu;XIA Ying-zi;LIN Shen;LIU Xin-cong;CHENG Ya-zhi;YU Wen;GAO Wen-xia;WU Sheng-xin(College of Agriculture,Fujian Agriculture and Forestry University/Fujian Key Laboratory of Crop Breeding by Design,Fuzhou 350002,China;Fujian Tobacco Monopoly Administration Institute of Tobacco Science,Fuzhou 350003,China)

机构地区：[1]福建农林大学农学院/福建省作物设计育种重点实验室,福州350002 [2]福建省烟草专卖局/烟草科学研究所,福州350003

出　　处：《西南农业学报》2025年第2期341-349,共9页Southwest China Journal of Agricultural Sciences

基　　金：中国烟草总公司福建省公司科技计划项目(2019350000240135、2019350000240136)。

摘　　要：【目的】旨在从福州北峰烟草青枯病病圃中分离出强致病性青枯菌并明确其类型。【方法】以采自福州北峰烟草青枯病病圃的感病烟株的茎部为材料,采用划线培养法分离病原菌;采用青枯菌特异性引物对3株青枯菌(分别命名为FZ-01、FZ-02和FZ-03)进行PCR扩增以进行初步验证;采用伤根灌菌接种法对3株菌株进行致病性检测;采用分子手段对筛选到的强致病性菌株(FZ-01)进行演化型鉴定、16S rDNA序列分析和序列变种鉴定;采用伤根灌菌接种法对FZ-01的生理小种进行鉴定。【结果】病原菌PCR扩增产物大小280 bp,符合目标条带大小,初步证明其为青枯菌;致病性检测发现,3株菌株均有很强的致病性,对致病性最强的菌株FZ-01进行鉴定,结果表明,该菌株为演化型I型,生理小种1号、生化变种III,即亚洲分支菌株;16S rDNA分析表明,该菌株序列与坚果类土壤青枯菌(菌株号:APK76 MF973211.1)亲缘关系最近,达到70%;基于青枯菌内切葡聚糖酶基因(egl)的系统发育学分析结果表明,该菌株为青枯菌序列变种17。【结论】本研究分离到的病原菌为强致病性青枯菌,且属于常见类型。【Objective】The study aimed to isolate highly pathogenic Ralstonia solanacearum and determine its type from the tobacco bacterial wilt nursery in Beifeng Fuzhou.【Method】The stems of diseased tobacco plants collected from the tobacco bacterial wilt disease nursery in Beifeng were used as materials,and the pathogen was isolated by streak culture method.Specific primers for R.solanacearum were used to perform PCR amplification on three strains(named FZ-01,FZ-02 and FZ-03 respectively)for preliminary verification.The pathogenicity detection of the three strains was carried out by root injury and bacterial irrigation inoculation method.Molecular methods were used to identify the evolutionary type,16S rDNA sequence analysis and sequence variant identification of the highly pathogenic strain(FZ-01).The physiological races of FZ-01 were identified by root injury and bacterial irrigation inoculation method.【Result】The size of the PCR amplification product of the pathogen was 280 bp,which was in line with the target band size,preliminarily proving that it was R.solanacearum.The pathogenicity test showed that the three strains had strong pathogenicity,and the most pathogenic strain FZ-01 was identified,the identification results of strain FZ-01 showed that the strain was evolutionary type I,physiological race 1,and biochemical variant III,that was,an Asian branch strain.16S rDNA analysis showed that the sequence of this strain had the closest genetic relationship with R.solanacearum in nut soil(strain number:APK76 MF973211.1),reaching 70%.The R.solanacearum strain was classified as sequence variant 17 based on phylogenetic analysis of the egl gene.【Conclusion】The isolated pathogen is indeed a highly pathogenic R.solanacearum and belongs to a common type.

关 键 词：烟草 青枯菌 生理小种 生化变种 序列变种 16S rDNA 

分 类 号：S572[农业科学—烟草工业]