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Applied and Environmental Microbiology, October 2008, p. 5934-5942, Vol. 74, No. 19
0099-2240/08/$08.00+0     doi:10.1128/AEM.02602-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Crenarchaeota and Their Role in the Nitrogen Cycle in a Subsurface Radioactive Thermal Spring in the Austrian Central Alps{triangledown}

Gerhard W. Weidler, Friedrich W. Gerbl, and Helga Stan-Lotter*

Department of Molecular Biology, Division of Microbiology, University of Salzburg, Billrothstraße 11, A-5020 Salzburg, Austria

Received 17 November 2007/ Accepted 11 August 2008

Previous results from a 16S rRNA gene library analysis showed high diversity within the prokaryotic community of a subterranean radioactive thermal spring, the "Franz-Josef-Quelle" (FJQ) in Bad Gastein, Austria, as well as evidence for ammonia oxidation by crenarchaeota. This study reports further characterization of the community by denaturing gradient gel electrophoresis (DGGE) analysis, fluorescence in situ hybridization (FISH), and semiquantitative nitrification measurements. DGGE bands from three types of samples (filtered water, biofilms on glass slides, and naturally grown biofilms), including samples collected at two distinct times (January 2005 and July 2006), were analyzed. The archaeal community consisted mainly of Crenarchaeota of the soil-subsurface-freshwater group (group 1.1b) and showed a higher diversity than in the previous 16S rRNA gene library analysis, as was also found for crenarchaeal amoA genes. No bacterial amoA genes were detected. FISH analysis of biofilms indicated the presence of archaeal cells with an abundance of 5.3% (±4.5%) in the total 4',6-diamidino-2-phenylindole (DAPI)-stained community. Microcosm experiments of several weeks in duration showed a decline of ammonium that correlated with an increase of nitrite, the presence of crenarchaeal amoA genes, and the absence of bacterial amoA genes. The data suggested that only ammonia-oxidizing archaea (AOA) perform the first step of nitrification in this 45°C environment. The crenarchaeal amoA gene sequences grouped within a novel cluster of amoA sequences from the database, originating from geothermally influenced environments, for which we propose the designation "thermal spring" cluster and which may be older than most AOA from soils on earth.

* Corresponding author. Mailing address: Department of Microbiology, Division of Molecular Biology, University of Salzburg, Billrothstraße 11, A-5020 Salzburg, Austria. Phone: 43 662 8044 7210. Fax: 43 662 8044 7209. E-mail: helga.stan-lotter{at}sbg.ac.at

{triangledown} Published ahead of print on 22 August 2008.

Applied and Environmental Microbiology, October 2008, p. 5934-5942, Vol. 74, No. 19
0099-2240/08/$08.00+0     doi:10.1128/AEM.02602-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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