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Toward a Wolbachia Multilocus Sequence Typing System: Discrimination of Wolbachia Strains Present in Drosophila Species

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Abstract

Among the diverse maternally inherited symbionts in arthropods, Wolbachia are the most common and infect over 20% of all species. In a departure from traditional genotyping or phylogenetic methods relying on single Wolbachia genes, the present study represents an initial Multilocus Sequence Typing (MLST) analysis to discriminate closely related Wolbachia pipientis strains, and additional data on sequence diversity in Wolbachia. We report a new phylogenetic characterization of four genes (aspC, atpD, sucB, and pdhB), and provide an expanded analysis of markers described in previous studies (16S rDNA, ftsZ, groEL, dnaA, and gltA). MLST analysis of the bacterial strains present in 16 different Drosophila–Wolbachia associations detected four distinct clonal complexes that also corresponded to maximum-likelihood identified phylogenetic clades. Among the 16 associations analyzed, six could not be assigned to MLST clonal complexes and were also shown to be in conflict with relationships predicted by maximum-likelihood phylogenetic inferences. The results demonstrate the discriminatory power of MLST for identifying strains and clonal lineages of Wolbachia and provide a robust foundation for studying the ecology and evolution of this widespread endosymbiont.

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Acknowledgments

The authors would like to thank Laura Baldo for her constructive comments and suggestions. The D. simulans (wMa) and D. innubila (wDin) were kindly provided by Prof. Bill Ballard and Prof. John Jaenike, respectively. This work was partially supported by intramural funds of the University of Ioannina to K. Bourtzis, by grants to J.J. Wernegreen from the National Institutes of Health (R01 GM62626-01) and the NASA Astrobiology Institute (NNA04CC04A), and to J.H. Werren and J.J. Wernegreen from the National Science Foundation (EF-0328363). This work was performed in part while S.R. Bordenstein held a National Research Council Research Associateship Award at the Marine Biological Laboratory.

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Authors and Affiliations

  1. Department of Environmental and Natural Resources Management, University of Ioannina, 2 Seferi St., 30100, Agrinio, Greece

    Charalampos Paraskevopoulos & Kostas Bourtzis

  2. Josephine Bay Paul Center for Comparative Molecular Biology and Evolution, The Marine Biological Laboratory, Woods Hole, MA, 02543, USA

    Seth R. Bordenstein & Jennifer J. Wernegreen

  3. Department of Biology, University of Rochester, Rochester, NY, 14627, USA

    John H. Werren

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  1. Charalampos Paraskevopoulos
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  2. Seth R. Bordenstein
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  3. Jennifer J. Wernegreen
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  5. Kostas Bourtzis
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Correspondence to Kostas Bourtzis.

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Paraskevopoulos, C., Bordenstein, S.R., Wernegreen, J.J. et al. Toward a Wolbachia Multilocus Sequence Typing System: Discrimination of Wolbachia Strains Present in Drosophila Species. Curr Microbiol 53, 388–395 (2006). https://doi.org/10.1007/s00284-006-0054-1

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