樱桃冠瘿病病原的分离与鉴定  

Isolation and identification of pathogen of cherry crown gall disease

作　　者：冯瑛[1] 蔡宇良[1] 宛甜[1] 张姗姗[1] 张雪[1] 

机构地区：[1]西北农林科技大学园艺学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2013年第10期165-171,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：西安市科技攻关项目(NJ200903);国家樱桃产业计划项目(200903019)

摘　　要：【目的】对近几年西安灞桥地区甜樱桃发生的冠瘿病进行病原分离与鉴定,探讨其地上部发病机理,为防治樱桃冠瘿病奠定基础。【方法】从陕西西安樱桃冠瘿病发病树体取样,通过发病部位镜检、病原菌分离纯化、致病性检测、病原菌生理生化检测结合分子生物学方法鉴定病原菌。【结果】从樱桃枝条和根系上的幼嫩冠瘿瘤分离纯化出6株细菌和1株真菌;通过刺伤接种樱桃幼嫩枝条、胡萝卜圆盘及烟草茎部,编号为CG04的菌株能够产生幼瘤并可重新分离出来;将CG04菌株的16SrDNA序列在NCBI上进行BLAST(登录号:KC107786)比对,同源性高于98%的菌株均为土壤杆菌属Agrobacteriumspp.,结合生化指标鉴定CG04菌株为根癌农杆菌(Agrobacterium tumefaciens)生物Ⅰ型菌种。【结论】确定根癌农杆菌生物Ⅰ型是樱桃枝条和根产生冠瘿瘤的致病菌。【Objective】Based on the isolation and identification of pathogen of cherry crown gall disease,the mechanism of disease occurrence above the ground was investigated to improve the prevention and control of the disease.【Method】Pathogenicity test was conducted based on Koch’s postulate.Microscopical examination,pathogen isolation,pathogenicity test,and physiological and biochemical test combined with molecular biology methods were used to identify pathogen.【Result】6 strains of bacteria and 1 strain of fungus were isolated from the crown gall.In the pathogenicity test,carrot disk,tobacco stem and one year old cherry branch were inoculated with all isolates,the position inoculated with CG04 could grow tumor and the same strain could be isolated from the new tumor.16S rDNA sequences of strain CG04 were submitted to NCBI to conduct the BLAST （Registry Number：KC107786）,and the strains whose homology was higher than 98% were all Agrobacterium spp.CG04 was identified to be Agrobacterium tumefaciens （biological type Ⅰ strain） upon biochemical test and molecular result.【Conclusion】The identified pathogen of cherry crown gall disease was A.tumefaciens biological type Ⅰ strain.

关 键 词：樱桃 冠瘿病 根癌农杆菌 16S RDNA 

分 类 号：S436.629[农业科学—农业昆虫与害虫防治]