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http://dx.doi.org/10.5352/JLS.2012.22.2.245

Differentiation of Vibrio spp. including Core Group Species by PCR-RFLP  

Park, Jin-Sook (Department of Biotechnology, Hannam University)
Publication Information
Journal of Life Science / v.22, no.2, 2012 , pp. 245-250 More about this Journal
Abstract
The 16S rDNA - RFLP types for six Vibrio species (V. fluvialis, V. proteolyticus, V. vulnificus, V. mimicus) including two core group members, V. alginolyticus and V. parahaemolyticu s, and Grimontia (Vibrio) hollisae were determined using PCR-RFLP analysis. Six tetrameric restriction enzymes (Alu I, Cfo I, Dde I, Hae III, Msp I, and Rsa I) were selected for RFLP analysis. V. alginolyticus, V. parahaemolyticus, and V. proteolyticus showed the same RFLP pattern following digestion with four of the six used restriction enzymes: CfoI, DdeI, MspI, and RsaI. Various restriction enzyme combinations generated digests recognizable as distinct RFLP types for each of the assayed Vibrio species. In particular, AluI single digestion produced species specific band patterns that enabled the differentiation between these Vibrio species. Dendrogram based on restriction patterns showed that two Vibrio core group members, V. alginolyticus and V. parahaemolyticus were closely related having a similarity over 90%. Although the observed RFLP pattern for Grimontia hollisae shared several common bands with other Vibrio spp., G. hollisae results were still clearly distinct from Vibrio spp. RFLP types for all restriction enzymes tested. If restriction enzymes are aptly selected, PCR-RFLP analysis is still a rapid and effective tool for differentiating Vibrio species.
Keywords
16S rDNA; core group; PCR-RFLP; Vibrio;
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1 Thompson, F. L., B. Hoste, K. Vandemeulebroecke, and J. Swings. 2003. Reclassification of Vibrio hollisea as Grimontia hollisae gen. nov., comb. Nov. Inc. J. syst. Evol. Microbiol. 53, 1615-1617.   DOI   ScienceOn
2 Thompson, F. L., D. Gevers, C. C. Thompson, P. Dawyndt, S. Naser, B. Hoste, C. B. Munn, and J. Swings. 2005. Phylogeny and molecular identification of vibrios on the basis of multilocus sequence analysis. Appl. Microbiol. Biotechnol. 71, 5107-5115.
3 Thompson, F. L., T. Iida, and J. Swings. 2004. Biodiversity of vibrios. Microbiol. Mol. Biol. Rev. 68, 403-431.   DOI   ScienceOn
4 Urakawa, H., K. Kita-Tsukamoto, and K. Ohwada. 1997. 16S rDNA genotyping using PCR/RFLP (restriction fragment length polymorphism) analysis among the family Vibrionaceae. FEMS Microbiol. Lett. 152, 125-132.   DOI   ScienceOn
5 Urakawa, H., K. Kita-Tsukamoto, and K. Ohwada. 1998. A new approach to separate the genus Photobacterium from Vibrio with RFLP patterns by HhaI digestion of PCR-amplified 16S rDNA. Curr. Microbiol. 36, 171-174.   DOI   ScienceOn
6 Lee, J. W., I. J. Jun, H. J. Kwun, K. L. Jang, and J. H. Cha. 2004. Direct identification of Vibrio vulnificus by PCR targeting elastase gene. J. Microbiol. Biotechnol. 14, 284-289.
7 Lerat E, V. Daubin, and N. A. Moran. 2003. From gene trees to organismal phylogeny in prokaryotes: the case of the gammaproteobacteria. PLOS Biol. 1, E19.
8 Pascual, J., M. C. Macian, D. R. Arahal, E. Garay, and M. J. Pujalte. 2010. Multilocus sequence analysis of the central clade of the genus Vibrio by using the 16S rRNA, recA, pyrH, rpoD, gyrB, rctB and toxR genes. Inc. J. Syst. Evol. Microbiol. 60, 154-165.   DOI   ScienceOn
9 Pedersen, K., L. Verdonck, B. Austin, D. A. Austin, A. R. Blanch, P. A. D. Grimont, J. Jofre, S. Koblavi, J. L. Larsen, T. Tiainen, M. Vigneulle, and J. Swings. 1998. Taxonomic evidence tha Vibrio carchariae grimes et al. 1985 is a junior synonym of Vibrio harveyi (Johnson and Shunk 1936) Baumann et al. 1981. Int. J. Syst. Bacteriol. 48, 749-758.   DOI   ScienceOn
10 Rivas, R., P. Garcia-Fraile, P. F. Mateos, E. Martinez-Molina, and E. Velazquez. 2006. Photobacterium halotolerans sp. Nov., isolated from lake Martel in Spain. Inc. J. Syst. Evol. Microbiol. 56, 1067-1071.   DOI   ScienceOn
11 Kumar, N. R. and S. Nair. 2007. Vibrio rhizosphaerae sp. nov., a red-pigmented bacterium that antagonizes phytopathogenic bacteria. Inc. J. Syst. Evol. Microbiol. 57, 2241-2246.   DOI   ScienceOn
12 Yeon, S. H., W. J. Jeong, and J. S. Park. 2005. The diversity of culturable organotrophic bacteria from local solar salterns. J. Microbiol. 43, 1-10.
13 Ki, J. S., R. Zhang, W. Zhang, Y. L. Huang, and P. Y. Qian. 2009. Analysis of RNA polymerase beta subunit (rpoB) gene sequences for the discriminative power of marine Vibrio species. Microb Ecol. 58, 679-691.   DOI   ScienceOn
14 Kim, D. G., S. H. Ahn, L. H. Kim, K. J. Park, Y. K. Hong, and I. S. Kong. 2008. Application of the rpoS gene for species-specific detection of Vibrio vulnificus by real-time PCR. J. Microbiol. Biotechnol. 18, 1841-1847.
15 Lambert, M. A., F. W. Hickman-Brenner, J. J. FarmerIII, and C. W. Moss. 1983. Differentiation of Vibrionaceae species by their cellular fatty acid composition. Int. J. Syst. Bacteriol. 33, 777-792.   DOI
16 Gomez-Gil, B., F. L. Thompson, C. C. Thompson, A. Garcia-Gasca, A. Roque, and J. Swings. 2004. Vibrio hispanicus sp. nov., isolated from Artemia sp. and sea water in Spain. Inc. J. syst. Evol. Microbiol. 54, 261-265.   DOI   ScienceOn
17 Aznar, R., W. Ludwig., R. I. Amann, and K. H. Schlefer. 1994. Sequence determination of rRNA genes of pathogenic Vibrio species and whole-cell identification of Vibrio vulnificus with rRNA-targeted oligonucleotide probes. Int. J. Syst. Bacteriol. 44, 330-337.   DOI   ScienceOn
18 Dorsch, M., D. Lane, and E. Stackebrandt. 1992. Toward a phylogeny of the genus Vibrio based on 16S rRNA sequence. Int. J. Syst. Bacteriol. 42, 58-63.   DOI   ScienceOn
19 Gomez-Gil, B., S. Soto-Rodriguez, A. Garcia-Gasc, A. Roque, T. Vazquez-Juarez, F. L. Thompson, and J. Swings. 2004. Molecular identification of Vibrio harveyi-related isolates associated with diseased aquatic organisms. Microbiol. 150, 1769-1777.   DOI   ScienceOn
20 Hernandez, G. and J. Olmos. 2003. Molecular identification of pathogenic and nonpathogenic strains of Vibrio harveyi using PCR and RAPD. Appl. Microbiol. Biotechnol. 63, 722-727.   DOI
21 Ki, J. S. 2009. Heterogeneity analysis of the 16S rRNA gene sequence of the genus Vibrio. Korean J. Microbiol. 45, 430-434.