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Diseases of Aquatic Organisms

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DAO 129:71-83 (2018)  -  DOI:

Vibrio alginolyticus 16S-23S intergenic spacer region analysis, and PCR assay for identification of coral pathogenic strain XSBZ03

Hongyue Li1,2, Xiang Zhang1,2, Hao Long1, Chaoqun Hu3, Yongcan Zhou1,2, Shifeng Wang1,2, Shaowen Ke1,2, Zhenyu Xie1,2,*

1State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou 570228, PR China
2Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, Hainan University, Haikou 570228, PR China
3CAS Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, PR China
*Corresponding author:

ABSTRACT: 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: IGS0, IGSG, IGSIA, IGSAG, IGSGLV, and IGSGLAV. IGS0, IGSG, IGSIA, IGSAG and IGSGLV 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. IGSGLAV was found only in the HN08155 strain, a causative agent of fish disease. IGSGLAV, IGSGLV and IGSAG 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.

KEY WORDS: Porites andrewsi white syndrome · Vibrio alginolyticus · Intergenic spacer · IGS · Strain-specific primers · Diagnostic protocol

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Cite this article as: Li H, Zhang X, Long H, Hu C and others (2018) Vibrio alginolyticus 16S-23S intergenic spacer region analysis, and PCR assay for identification of coral pathogenic strain XSBZ03. Dis Aquat Org 129:71-83.

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