基于16S rRNA和recA香鱼鳗利斯顿氏菌的分离鉴定  

Isolation and identification of Listonella anguillarum from Plecoglossus altivelis based on 16S rRNA and recA

作　　者：王成义[1,2,3] 闫茂仓[2,3] 陈少波[2,3] 管敏鑫[1,4] 单乐州[2,3] 艾为明[1] 谢起浪[2,3] 蔡延(马奔) 

机构地区：[1]温州医学院生命科学学院 [2]浙江省海洋水产养殖研究所 [3]浙江省近岸水域生物资源开发与保护重点实验室 [4]Division of Human Genetics and Center for Hearing and Deafness Research,Cincinnati Children’s Hospital Medical Center,Cincinnati,Ohio 45229,USA [5]温州市工业科学研究院

出　　处：《海洋通报》2010年第1期84-90,共7页Marine Science Bulletin

基　　金：浙江省科技厅科研院所专项公益技术攻关项目(2007F10009);浙江省科技厅科研院所青年人才计划项目(2006R20001);温州市科技兴海项目(S20070051)

摘　　要：为研究引起香鱼(Plecoglossus altivelis)出血溃烂症病因及致病菌系统发育地位,本研究从患病香鱼的肝脏、肾脏及体表分离到11株病原菌(编号:X0901-X0911),运用常规细菌生理生化方法鉴定致病菌所属种类;运用16SrRNA基因、recA基因序列分析方法研究致病菌的系统发育地位。细菌生理生化鉴定结果表明:致病菌为鳗利斯顿氏菌,11株细菌生理生化结果相同,均为革兰氏阴性杆菌、氧化酶阳性、接触酶阳性、吲哚阳性、精氨酸脱羧酶阳性、精氨酸双水解酶阳性、硝酸盐还原阳性、甘露醇阳性、MR测定阳性;H2S阴性、V-P测定阴性等。根据16SrRNA基因、recA基因序列分别构建弧菌科常见细菌系统进化树,结果表明:致病菌与鳗利斯顿氏菌(Listonella anguillarum)均聚为一枝,聚类结果与细菌生理生化鉴定结果相符。致病菌与鳗利斯顿氏菌16SrRNA基因、recA基因的同源性分别为99.9%、99.8%。以recA基因构建的系统进化树的拓扑学结构与16SrRNA基因建树结果相类似。此外,与16SrRNA基因相比,recA基因在不同物种之间具有更高的鉴别能力,本研究表明recA基因适合作为弧菌科常见细菌物种间进化关系研究的标记。In order to investigate the pathogen of bleeding ulcers of Plecoglossus altivelis and the phylogenetic position of this pathogen, 1 lpathogen were isolated from liver, kidney and surface of diseased Plecoglossus altivelis with the NO. from X0901 to X0911. Routine bacterial identification was used to identify this pathogen and the phylogenetic position was investigated using 16S rRNA gene and recA gene sequences analysis. The results of physiological and biochemical tests suggested that they were gram-negative bacteria, and the results of oxidase, contact enzyme, producing indole, arginine decarboxylase, arginine dihydrolase, nitrate reduction tests and MR tests were positive; but they couldn＇t generate H2S and the results of V-P test were negative too. Molecular phylogenetic trees of common bacterial were constructed and the results suggested that the pathogen and Listonella anguillarum clustered with each other. And the results corresponded with that of routine bacterial identification. The identity of 16S rRNA gene and recA gene between pathogen and Listonella anguillarum were 99.9% and 99.8% respectively. The topology of phylogenetic tree constructed by recA gene was generally agreed with that of 16S rRNA gene. And recA gene was much more discriminatory than 16S rRNA gene. So the conclusion was recA gene was an alternative phylogenetic and identification marker for the common vibrios.

关 键 词：香鱼 鳗利斯顿氏菌 分离 鉴定 

分 类 号：P736.221[天文地球—海洋地质]