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Updating the taxonomic toolbox: classification of Alteromonas spp. using multilocus phylogenetic analysis and MALDI-TOF mass spectrometry

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Abstract

Bacteria of the genus Alteromonas are Gram-negative, strictly aerobic, motile, heterotrophic marine bacteria known for their versatile metabolic activities. Identification and classification of novel species belonging to the genus Alteromonas generally involves DNA–DNA hybridization (DDH) as distinct species often fail to be resolved at the 97 % threshold value of the 16S rRNA gene sequence similarity. In this study, the applicability of Multilocus Phylogenetic Analysis (MLPA) and Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) for the differentiation of Alteromonas species has been evaluated. Phylogenetic analysis incorporating five house-keeping genes (dnaK, sucC, rpoB, gyrB, and rpoD) revealed a threshold value of 98.9 % that could be considered as the species cut-off value for the delineation of Alteromonas spp. MALDI-TOF MS data analysis reconfirmed the Alteromonas species clustering. MLPA and MALDI-TOF MS both generated data that were comparable to that of the 16S rRNA gene sequence analysis and may be considered as useful complementary techniques for the description of new Alteromonas species.

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Acknowledgments

We would like to acknowledge Bio21 Institute for the MALDI-TOF MS access and partial support from RFBR 11-04-00781-a. All authors declare that no potential conflicts of interests exist.

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Authors and Affiliations

  1. Faculty of Life and Social Sciences, Swinburne University of Technology, PO Box 218, Hawthorn, Vic 3122, Australia

    Hooi Jun Ng, Hayden K. Webb, Russell J. Crawford, François Malherbe & Elena P. Ivanova

  2. Bioscreen, Bio21 Institute, The University of Melbourne, Melbourne, Vic 3010, Australia

    Henry Butt & Rachel Knight

  3. G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences, Vladivostok, 690022, Russia

    Valery V. Mikhailov

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  1. Hooi Jun Ng
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  2. Hayden K. Webb
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  3. Russell J. Crawford
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  4. François Malherbe
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  5. Henry Butt
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  7. Valery V. Mikhailov
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  8. Elena P. Ivanova
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Correspondence to Elena P. Ivanova.

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Ng, H.J., Webb, H.K., Crawford, R.J. et al. Updating the taxonomic toolbox: classification of Alteromonas spp. using multilocus phylogenetic analysis and MALDI-TOF mass spectrometry. Antonie van Leeuwenhoek 103, 265–275 (2013). https://doi.org/10.1007/s10482-012-9807-y

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