期刊
DISEASES OF AQUATIC ORGANISMS
卷 129, 期 1, 页码 71-83出版社
INTER-RESEARCH
DOI: 10.3354/dao03233
关键词
Porites andrewsi white syndrome; Vibrio alginolyticus; Intergenic spacer; IGS; Strain-specific primers; Diagnostic protocol
资金
- National Natural Science Foundation of China [41466002, 31060360, 30760190]
- Fok Ying-Tong Education Foundation for Young Teachers in the Higher Education Institutions of China [121030]
- Major Science and Technology Project of Hainan Province [ZDZX2013014]
- Natural Science Foundation of Hainan Province [311031]
Porites andrewsi white syndrome (PAWS), caused by Vibrio alginolyticus strains XSBZ03 and XSBZ14, poses a serious threat to corals in the South China Sea. To obtain a specific target against which to develop a rapid PCR detection method for the coral pathogenic strain XSBZ03, the 16S-23S rRNA gene intergenic spacer (IGS) region of 4 strains of V. alginolyticus, including the XSBZ03 and XSBZ14 strains, was amplified, sequenced and analyzed. Six types of IGS were found: IGS(0), IGS(G), IGS(IA), IGS(AG), IGS(GLV), and IGS(GLAV). IGS(0), IGS(G), IGS(IA), IGS(AG) and IGS(GLV) appeared to be the most prevalent forms in the 4 strains and the percentage identity range within each type was 91.4-100%, 89.3-98.5%, 83.0-99.8%, 91.5-95.6%, and 88.7-99.3%, respectively. IGS(GLAV) was found only in the HN08155 strain, a causative agent of fish disease. IGS(GLAV), IGS(GLV) and IGS(AG) are reported here for the first time in V. alginolyticus. An IGS sequence specific to the XSBZ03 strain was identified following alignment of the homologous IGSs, and used to design strain-specific primers for its rapid identification by PCR. The results from PCR analysis suggest that the method is a rapid, practical, and reliable tool for the identification of the XSBZ03 strain in samples of isolated bacteria, as well as seawater and coral samples spiked with the bacterial strain. This is the first report of a rapid diagnostic assay for a causative agent of PAWS, based on PCR detection of a coral pathogen at the strain level. After applying this assay in coral transplantation, the survival rates of transplanted corals were significantly increased. This diagnostic assay should aid with both the elucidation of the cause of the disease, and transplantation of PAWS-free P. andrewsi in the South China Sea.
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