扩增核糖体DNA限制性分析(ARDRA)研究刺芹侧耳细菌病原菌  

Identification of Bacterial Pathogens of Pleurotus eryngii by ARDRA

作　　者：唐利华[1] 高君辉[1] 尚晓冬[1] 章炉军[1] 李玉[1] 谭琦[1] 

机构地区：[1]上海市农业科学院食用菌研究所,农业部南方食用菌资源利用重点实验室,国家食用菌工程技术研究中心,国家食用菌加工技术研发分中心,上海市农业遗传育种重点开放实验室,上海201403

出　　处：《食用菌学报》2012年第1期70-74,共5页Acta Edulis Fungi

基　　金：上海市农业科学院青年基金[编号:农青年科技2008(05)]的部分研究内容

摘　　要：为分析刺芹侧耳(Pleurotus eryngii)工厂化栽培中发黄腐烂子实体的细菌组成,构建16SrDNA文库,利用限制性内切酶MspI和RsaI分别对随机克隆进行酶切筛选,并测定代表性克隆16SrDNA序列,BLAST分析结果表明刺芹侧耳子实体病状组织中存有两类菌群,分别为假单胞菌和芽孢杆菌属。同时,对刺芹侧耳子实体病状组织中的细菌进行分离培养、致病性测定、16SrDNA鉴定及BLAST分析,病原细菌与恶臭假单胞菌(Pseudomonas putida)的同源性较高,其作为引起刺芹侧耳工厂化栽培细菌性病害的病原菌在国内外尚属首次报道。Rotted yellow fruit body tissue of Pleurotus eryngii obtained from an industrial mushroom cultivation facility was used to create a bacterial 16S rDNA library.Random clones were screened and obtained using the restriction endonucleases MspI and RsaI,and the 16S rDNA sequences of representative clones were identified.A BLAST search revealed the presence of two bacterial types in the disease tissue,which exhibited 97% and 93% similarity with Pseudomonas and Bacillus.Bacterial isolation and back inoculation on to P.eryngii primordia demonstrated that three isolates（designated T1,T2 and T3） produced symptoms on mushroom fruit bodies that were similar to the original diseased tissue.Comparison with known 16S rDNA sequences revealed that these pathogenic bacteria exhibited a high homology with Pseudomonas putida.

关 键 词：刺芹侧耳 工厂化栽培 细菌病害 克隆文库 16S rDNA基因序列 

分 类 号：S436.461[农业科学—农业昆虫与害虫防治]