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Spatial distribution of nitrogen fixation in methane seep sediment and the role of the ANME archaea

Dekas, Anne E. and Chadwick, Grayson L. and Bowles, Marshall W. and Joye, Samantha B. and Orphan, Victoria J. (2014) Spatial distribution of nitrogen fixation in methane seep sediment and the role of the ANME archaea. Environmental Microbiology, 16 (10). pp. 3012-3029. ISSN 1462-2912.

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Nitrogen (N_2) fixation was investigated at Mound 12, Costa Rica, to determine its spatial distribution and biogeochemical controls in deep-sea methane seep sediment. Using ^(15)N_2 tracer experiments and isotope ratio mass spectrometry analysis, we observed that seep N_2 fixation is methane-dependent, and that N_2 fixation rates peak in a narrow sediment depth horizon corresponding to increased abundance of aggregates of anaerobic methanotrophic archaea (ANME-2) and sulfate-reducing bacteria (SRB). Using fluorescence in situ hybridization coupled to nanoscale secondary ion mass spectrometry (FISH-NanoSIMS), we directly measured ^(15)N_2 uptake by ANME-2/SRB aggregates (n = 26) and observed maximum ^(15)N incorporation within ANME-2-dominated areas of the aggregates, consistent with previous analyses. NanoSIMS analysis of single cells (n = 34) from the same microcosm experiment revealed no ^(15)N_2 uptake. Together, these observations suggest that ANME-2, and possibly physically associated SRB, mediate the majority of new nitrogen production within the seep ecosystem. ANME-2 diazotrophy was observed while in association with members of two distinct orders of SRB: Desulfobacteraceae and Desulfobulbaceae. The rate of N_2 fixation per unit volume biomass was independent of the identity of the associated SRB, aggregate size and morphology. Our results show that the distribution of seep N_2 fixation is heterogeneous, laterally and with depth in the sediment, and is likely influenced by chemical gradients affecting the abundance and activity of ANME-2/SRB aggregates.

Item Type:Article
Related URLs:
URLURL TypeDescription DOIArticle Information
Dekas, Anne E.0000-0001-9548-8413
Chadwick, Grayson L.0000-0003-0700-9350
Bowles, Marshall W.0000-0002-0464-5880
Joye, Samantha B.0000-0003-1610-451X
Orphan, Victoria J.0000-0002-5374-6178
Additional Information:© 2013 Society for Applied Microbiology and John Wiley & Sons Ltd. Received 11 February, 2013; revised 24 May, 2013; accepted 9 August, 2013. Article first published online: 24 SEP 2013. We would like to thank Lisa Levin, Burt Thomas, David Fike, Jake Bailey, Rachel Poretsky, Shana Goffredi and the crew and science parties of AT 15–44 and AT 15–59 for assistance in sample collection. Yunbin Guan and John Eiler are acknowledged for assistance with NanoSIMS operations, Bill Ussler (Monterey Bay Aquarium Research Institute) and Nathan Dalleska (Caltech Environmental Analysis Center) for assistance with methane and ammonium measurements, Steve Skinner for map construction, and Stephanie Connon and Abigail Green-Saxena for assistance with the sulfide measurements and CARD-FISH respectively. We are also grateful to Abigail Green-Saxena and Roland Hatzenpichler for detailed reviews of this manuscript. We would also like to thank three anonymous reviewers for their careful consideration and suggested improvements to the manuscript. Funding was provided by the Department of Energy Biological & Environmental Research (BER) (Early Career Award DE-SC0003940 and DE-SC0004949 to V.J.O) and the National Science Foundation (MCB-0348492 to V.J.O and EF-0801741 to S.B.J.) and an NSF Graduate Research Fellowship to A.E.D. The editing of this work by A.E.D. was partially performed under the auspices of the US Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. The Caltech Center for Microanalysis, housing the CAMECA NanoSIMS 50L, is funded in part by the Gordon and Betty Moore Foundation.
Funding AgencyGrant Number
Department of Energy (DOE)DE-SC0003940
Department of Energy (DOE)DE-SC0004949
NSF Graduate Research FellowshipUNSPECIFIED
Department of Energy (DOE)DE-AC52-07NA27344
Gordon and Betty Moore FoundationUNSPECIFIED
Issue or Number:10
Record Number:CaltechAUTHORS:20141201-133733752
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Official Citation:Dekas, A. E., Chadwick, G. L., Bowles, M. W., Joye, S. B. and Orphan, V. J. (2014), Spatial distribution of nitrogen fixation in methane seep sediment and the role of the ANME archaea. Environmental Microbiology, 16: 3012–3029. doi: 10.1111/1462-2920.12247
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:52235
Deposited By: Tony Diaz
Deposited On:01 Dec 2014 21:51
Last Modified:03 Oct 2019 19:05

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