中国对虾来源哈维氏弧菌鉴定及生物学分析

doi:10.11924/j.issn.1000-6850.casb17050026

中国农学通报 ›› 2019, Vol. 35 ›› Issue (3): 34-39.doi: 10.11924/j.issn.1000-6850.casb17050026

所属专题：水产渔业

中国对虾来源哈维氏弧菌鉴定及生物学分析

樊英

山东省海洋生物研究院

收稿日期:2017-05-05 修回日期:2018-12-27 接受日期:2017-10-16 出版日期:2019-01-28 发布日期:2019-01-28
通讯作者: 樊英
基金资助:
国家现代农业产业技术体系建设专项资助资金(CARS-47)；山东省现代农业产业技术体系虾蟹类产业创新团队(SDAIT-13)。

Identification and Molecular Characteristics of Vibrio harveyi from Fenneropenaeus chinensis

Received:2017-05-05 Revised:2018-12-27 Accepted:2017-10-16 Online:2019-01-28 Published:2019-01-28

摘要/Abstract

摘要： 从患病中国对虾中分离到优势菌株,对其进行生理生化特性及分子生物学分析。TCBS培养基上菌落呈现绿色(以下称为TCBS-G), 通过形态学、生理生化分析,菌株TCBS-G为革兰氏阴性菌,短杆状,两端钝圆,有鞭毛；Biolog GIII分析表明,该菌株生长代谢利用情况与弧菌吻合,与哈维氏弧菌的相似系数为？。基于16S rRNA、gyrB、HSP60基因序列分析同源性,16S rRNA未将TCBS-G有效鉴定,gyrB、HSP60基因序列与哈维氏弧菌相聚类,聚合置信度可达99%。综合生物学及分子特征,判定TCBS-G为弧菌属(Vibrio pacini)的哈维氏弧菌(Vibrio harveyi)。底物平板试验结果显示TCBS-G菌株胞外产物具有淀粉酶、蛋白酶、脂肪酶和DNA酶活性；且能够扩增出哈氏弧菌溶血素基因的保守特异性片段。

关键词: 芒草, 芒草, ISSR-PCR反应体系, 单因子试验, 引物筛选

Abstract: In this study, the dominant strain was isolated from diseased Fenneropenaeus chinensis , the biochemical and molecular biological identification were conducted for this strain. The strain was green on TCBS medium (thus designated as TCBS-G). Based on physiological and biochemical analysis, TCBS-G was identified to be Gram-negative, in short rod shape, blunt ends, and with flagellum. Biolog GIII experiment showed that the growth and metabolism of TCBS-G were similar to that ofVibrio harveyi . Homology of TCBSG withVibrio spp. was analyzed based on the sequences of 16S rRNA gene,gyrB andHSP60 . Based on the sequence of 16S rRNA gene, TCBS-G was not identified. Phylogenetic tree based ongyrB andHSP60 genes showed that TCBS-G clustered together withV. harveyi , with a high bootstrap value of 99%. In summary, all above results confirmed that the isolated strain TCBS-G wasV. harveyi . The substrate plate test showed that it could produce diastase, proteinase, lipase, and DNase. In addition, the conserved region in hemolysin gene of V. harveyi could be amplified by PCR with specific primers.

樊英. 中国对虾来源哈维氏弧菌鉴定及生物学分析[J]. 中国农学通报, 2019, 35(3): 34-39.

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中国对虾来源哈维氏弧菌鉴定及生物学分析

Identification and Molecular Characteristics of Vibrio harveyi from Fenneropenaeus chinensis

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