猕猴桃溃疡病病原菌的分子鉴定和致病力测定  

Moleculer identification and pathogenicity detection of Pseudomonas syringae pv.actinidae

作　　者：赵利娜[1] 胡家勇[1] 叶振风[1] 刘普[1] 朱立武[1] 

机构地区：[1]安徽农业大学果树学重点实验室,合肥230036

出　　处：《华中农业大学学报》2012年第5期604-608,共5页Journal of Huazhong Agricultural University

基　　金：安徽省重点科研计划项目(07020303049)

摘　　要：为明确猕猴桃细菌性溃疡病致病菌种类与特征,以从安徽岳西、陕西户县和重庆黔江等猕猴桃主产区收集到的溃疡病感染枝条和树皮为材料,采用平板划线、梯度稀释和BPA培养法分离病原物,采用烟草过敏性反应和猕猴桃健康叶片接种观察其致病性,结合特异引物扩增的16S-23SrDNA-ITS序列分析,对病原菌种类进行分子鉴定。结果表明:从收集的溃疡病感染枝条和树皮中,共分离纯化得到10株分离物;所有分离物均可使烟草叶片产生过敏反应,猕猴桃叶片接种显示同一菌株对不同品种及不同菌株对同一品种的致病力均存在差异。对特异扩增得到的280bp 16S-23SrDNA-ITS序列进行Blast比对分析,结果表明:10个菌株为同一致病菌,其DNA片段大小和序列均与丁香假单孢杆菌猕猴桃致病变种(Pseudomonas syringae pv.actinidae)完全一致。据此可以确定,试验分离所得的菌株均为丁香假单孢杆菌猕猴桃致病变种。To identify the pathogenic bacteria of kiwifruit bacterial canker, the infected kiwifruit branches and barks from Anhui （Yuexi County） Province, Shaan＇ xi （Huxian County） Province and Chongqing （Qianjiang County） Municipality were collected as the experiment materials. Plate streak- ing, gradient dilution and beef extract peptone agar （BPA） media cultivation were used for bacteria iso- lation, and tobacco hypersensitive responses detection, kiwifruit pathogenicity detection and 16S-23S rDNA-ITS sequential analysis were carried out as follows for bacteria identification. From the bacterial canker infected kiwifruit tissues, 10 pathogenic strains were isolated, and all isolated strains have patho- genicity and hypersensitivity to tested kiwifruit and tobacco leaves. However, different isolated strains have different pathogenicity for kiwifruit. 10 purified pathogenic strains belonged to the same pathogen through 16S-23S rDNA-ITS and Blast sequential analysis. The amplified 280 bp sequences were the same as the previously reported 16S-23S rDNA-ITS of Pseudomonas syringae pv. actinidae. It is con- cluded that the isolates from the canker infected kiwifruit branches and barks were Pseudomonas syrin- gae pv. actinidae.

关 键 词：猕猴桃 溃疡病 丁香假单孢杆菌 鉴定 致病力 

分 类 号：S436.634.12[农业科学—农业昆虫与害虫防治] S663.4[农业科学—植物保护]