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Sulfur cycling and methanogenesis primarily drive microbial colonization of the highly sulfidic Urania deep hypersaline basin
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Edited by David M. Karl, University of Hawaii, Honolulu, HI, and approved April 14, 2009 (received for review November 25, 2008)

Abstract
Urania basin in the deep Mediterranean Sea houses a lake that is >100 m deep, devoid of oxygen, 6 times more saline than seawater, and has very high levels of methane and particularly sulfide (up to 16 mM), making it among the most sulfidic water bodies on Earth. Along the depth profile there are 2 chemoclines, a steep one with the overlying oxic seawater, and another between anoxic brines of different density, where gradients of salinity, electron donors and acceptors occur. To identify and differentiate the microbes and processes contributing to the turnover of organic matter and sulfide along the water column, these chemoclines were sampled at a high resolution. Bacterial cell numbers increased up to a hundredfold in the chemoclines as a consequence of elevated nutrient availability, with higher numbers in the upper interface where redox gradient was steeper. Bacterial and archaeal communities, analyzed by DNA fingerprinting, 16S rRNA gene libraries, activity measurements, and cultivation, were highly stratified and metabolically more active along the chemoclines compared with seawater or the uniformly hypersaline brines. Detailed analysis of 16S rRNA gene sequences revealed that in both chemoclines δ- and ε-Proteobacteria, predominantly sulfate reducers and sulfur oxidizers, respectively, were the dominant bacteria. In the deepest layers of the basin MSBL1, putatively responsible for methanogenesis, dominated among archaea. The data suggest that the complex microbial community is adapted to the basin's extreme chemistry, and the elevated biomass is driven largely by sulfur cycling and methanogenesis.
- deep anoxic hypersaline lake
- element cycling
- geosphere–biosphere interaction
- Mediterranean Sea
- microbial diversity
Footnotes
- 1To whom correspondence should be addressed at: Department of Food Science and Microbiology, Università degli Studi di Milano, Via Celoria 2, 20133 Milan, Italy. E-mail: daniele.daffonchio{at}unimi.it
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Author contributions: S.B. and D.D. designed research; S.B., L.B., F.M., G.D., T.B., A.R., M.Y., D.M., G.J.D.L., P.V.d.W., E.M., C.C., and D.D. performed research; S.B., L.B., F.M., G.D., M.M., M.Y., D.M., G.J.D.L., H.B., T.J.M., P.N.P., E.M., L.G., C.C., and D.D. analyzed data; and S.B., T.J.M., and D.D. wrote the paper.
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The authors declare no conflict of interest.
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This article is a PNAS Direct Submission.
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Data deposition: 16S rRNA sequences have been submitted to the GenBank database (accession nos. AY164322–AY164333, AY164429–AY164455, AY226324–AY226340, AY226377–AY226381, AY547867–AY548016, and DQ453257–DQ453476).
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This article contains supporting information online at www.pnas.org/cgi/content/full/0811984106/DCSupplemental.
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Freely available online through the PNAS open access option.
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