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Applied and Environmental Microbiology, February 2006, p. 1364-1372, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1364-1372.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification of a Thiomicrospira denitrificans-Like Epsilonproteobacterium as a Catalyst for Autotrophic Denitrification in the Central Baltic Sea

Ingrid Brettar,¹ Matthias Labrenz,^1, Sébastien Flavier,² Julia Bötel,¹ Harri Kuosa,³ Richard Christen,² and Manfred G. Höfle^1*

GBF-German Research Center for Biotechnology, Department of Environmental Microbiology, Mascheroder Weg 1, D-38124 Braunschweig, Germany,¹ UMR6543 CNRS-Université de Nice Sophia Antipolis, Centre de Biochimie, Parc Valrose, F-06108 Nice, France,² Tvärminne Zoological Station, FI-10900 Hanko, Finland³

Received 7 April 2005/ Accepted 28 November 2005

Identification and functional analysis of key members of bacterial communities in marine and estuarine environments are major challenges for obtaining a mechanistic understanding of biogeochemical processes. In the Baltic Sea basins, as in many other marine environments with anoxic bodies of water, the oxic-anoxic interface is considered a layer of high bacterial turnover of sulfur, nitrogen, and carbon compounds that has a great impact on matter balances in the whole ecosystem. We focused on autotrophic denitrification by oxidation of reduced sulfur compounds as a biogeochemically important process mediating concomitant turnover of sulfur, nitrogen, and carbon. We used a newly developed approach consisting of molecular analyses in stimulation experiments and in situ abundance. The molecular approach was based on single-strand conformational polymorphism (SSCP) analysis of the bacterial community RNA, which allowed identification of potential denitrifiers based on the sequences of enhanced SSCP bands and monitoring of the overall bacterial community during the experiments. Sequences of the SSCP bands of interest were used to design highly specific primers that enabled (i) generation of almost complete 16S rRNA gene sequences using experimental and environmental DNA as templates and (ii) quantification of the bacteria of interest by real-time PCR. By using this approach we identified the bacteria responsible for autotrophic denitrification as a single taxon, an epsilonproteobacterium related to the autotrophic denitrifier Thiomicrospira denitrificans. This finding was confirmed by material balances in the experiments that were consistent with those obtained with continuous cultures of T. denitrificans. The presence and activity of a bacterium that is phylogenetically and physiologically closely related to T. denitrificans could be relevant for the carbon budget of the central Baltic Sea because T. denitrificans exhibits only one-half the efficiency for carbon dioxide fixation per mol of sulfide oxidized and mol of nitrate reduced of Thiobacillus denitrificans hypothesized previously for this function.

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* Corresponding author. Mailing address: GBF-German Research Center for Biotechnology, Department of Environmental Microbiology, Mascheroder Weg 1, D-38124 Braunschweig, Germany. Phone: 49 531 6181 419. Fax: 49 531 6181 411. E-mail: mho{at}gbf.de.

Supplemental material for this article may be found at http://aem.asm.org/.

Present address: IOW-Institute for Baltic Sea Research Warnemünde, Seestr. 15, D-18119 Rostock-Warnemünde, Germany.

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Applied and Environmental Microbiology, February 2006, p. 1364-1372, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1364-1372.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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