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Development of a PCR method for detection of Pseudoalteromonas marina associated with green spot disease in Pyropia yezoensis

文献类型： 外文期刊

作者： Yang, Huichao 1 ; Yan, Yongwei 1 ; Li, Jie 1 ; Tang, Lei 3 ; Mao, Yunxiang 3 ; Mo, Zhaolan 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Lab Marine Fisheries Sci & Food Prod Proc,Yellow, Qingdao Natl Lab Marine Sci & Technol,Qingdao Key, Key Lab Maricultural Organism Dis Control,Minist, Qingdao 266071, Peoples R China

2.Shanghai Ocean Univ, Coll Fisheries & Life Sci, Shanghai 201306, Peoples R China

3.Ocean Univ China, Lab Marine Biol & Biotechnol, Qingdao Natl Lab Marine Sci & Technol, Qingdao 266003, Peoples R China

4.Ocean Univ China, Key Lab Marine Genet & Breeding, Minist Educ, Qingdao 266003, Peoples R China

关键词： Pyropia yezoensis; green spot disease (GSD); Pseudoalteromonas marina; PCR detection; early diagnosis

期刊名称：JOURNAL OF OCEANOLOGY AND LIMNOLOGY （ 影响因子：1.265； 五年影响因子：1.276 ）

ISSN： 2096-5508

年卷期： 2020 年 38 卷 1 期

页码：

收录情况： SCI

摘要： Pseudoalteromonas marina is one of the potential pathogens that cause green spot disease (GSD) in Pyropia yezoensis. To prevent GSD from development and spread, an effective method to detect the pathogen at early GSD infection stages need to be established. In this research, PCR methods were established targeting the dnaA gene (encoding chromosome replication initiator protein) and the dnaN gene (encoding beta sliding clamp of DNA polymerase III protein) to detect P. marina with three primer pairs pws-dnaA2 (Forward, 5 '-ACCGCATTAACGAACTACTCGTG-3 '; Reverse, 5 '-TGCCATTACCTACAGCATGG-3 '), pcs-dnaN2 (Forward, 5 '-CTTACAACGTTATCAGCGGC-3 '; Reverse, 5 '-GTTGAGTATTAAGTGATTGAGTAAGC-3 ') or pws-dnaN3 (Forward, 5 '-ACTTACAACGTTATCAGCGGC-3 '; Reverse, 5 '-ACTGCTGTTTGAGTCTGCTAAC-3 '). Three PCR methods corresponding to the three primer pairs sufficiently distinguished P. marina from 22 bacterial species, thus resulting in detection limits of 4 to 4x10(2) CFU cells or 2.37x10(1) to 2.37x10(3) fg of P. marina DNA per PCR reaction. In an artificial infection experiment of P. yezoensis infected with P. marina, all established PCRs successfully detected P. marina at early GSD infection stages. The results show that the established PCRs are specific and sensitive, and are potential for applications in early diagnosis of GSD in Pyropia.

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