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Applied and Environmental Microbiology, September 2005, p. 5107-5115, Vol. 71, No. 9
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.9.5107-5115.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Phylogeny and Molecular Identification of Vibrios on the Basis of Multilocus Sequence Analysis

F. L. Thompson,1* D. Gevers,2 C. C. Thompson,1 P. Dawyndt,2 S. Naser,2 B. Hoste,2 C. B. Munn,3 and J. Swings2

Microbial Resources Division and Brazilian Collection of Environmental and Industrial Micro-Organisms (CBMAI), CPQBA, UNICAMP, CP 6171, 13081-970 Campinas, Brazil,1 Laboratory of Microbiology and BCCM/LMG Bacteria Collection, Ghent University, K. L. Ledeganckstraat 35, Ghent 9000, Belgium,2 School of Biological Sciences, University of Plymouth, B417, Portland Square, Drake Circus, Plymouth, Devon PL4 8AA, United Kingdom3

Received 9 August 2004/ Accepted 3 April 2005

We analyzed the usefulness of rpoA, recA, and pyrH gene sequences for the identification of vibrios. We sequenced fragments of these loci from a collection of 208 representative strains, including 192 well-documented Vibrionaceae strains and 16 presumptive Vibrio isolates associated with coral bleaching. In order to determine the intraspecies variation among the three loci, we included several representative strains per species. The phylogenetic trees constructed with the different genetic loci were roughly in agreement with former polyphasic taxonomic studies, including the 16S rRNA-based phylogeny of vibrios. The families Vibrionaceae, Photobacteriaceae, Enterovibrionaceae, and Salinivibrionaceae were all differentiated on the basis of each genetic locus. Each species clearly formed separated clusters with at least 98, 94, and 94% rpoA, recA, and pyrH gene sequence similarity, respectively. The genus Vibrio was heterogeneous and polyphyletic, with Vibrio fischeri, V. logei, and V. wodanis grouping closer to the Photobacterium genus. V. halioticoli-, V. harveyi-, V. splendidus-, and V. tubiashii-related species formed groups within the genus Vibrio. Overall, the three genetic loci were more discriminatory among species than were 16S rRNA sequences. In some cases, e.g., within the V. splendidus and V. tubiashii group, rpoA gene sequences were slightly less discriminatory than recA and pyrH sequences. In these cases, the combination of several loci will yield the most robust identification. We can conclude that strains of the same species will have at least 98, 94, and 94% rpoA, recA, and pyrH gene sequence similarity, respectively.

* Corresponding author. Mailing address: Microbial Resources Division and Brazilian Collection of Environmental and Industrial Micro-Organisms (CBMAI), CPQBA, UNICAMP, CP 6171, 13081-970 Campinas, Brazil. Phone: 55-19-38847500. Fax: 55-19-38847811. E-mail: Fabiano.Thompson{at}terra.com.br.

Applied and Environmental Microbiology, September 2005, p. 5107-5115, Vol. 71, No. 9
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.9.5107-5115.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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