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Patterns of ^(15)N assimilation and growth of methanotrophic ANME-2 archaea and sulfate-reducing bacteria within structured syntrophic consortia revealed by FISH-SIMS

Orphan, Victoria J. and Turk, Kendra A. and Green, Abigail M. and House, Christopher H. (2009) Patterns of ^(15)N assimilation and growth of methanotrophic ANME-2 archaea and sulfate-reducing bacteria within structured syntrophic consortia revealed by FISH-SIMS. Environmental Microbiology, 11 (7). pp. 1777-1791. ISSN 1462-2912.

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Methane release from the oceans is controlled in large part by syntrophic interactions between anaerobic methanotrophic archaea (ANME) and sulfate-reducing bacteria (DSS), frequently found as organized consortia. An understanding of the specifics of this symbiotic relationship and the metabolic heterogeneity existing between and within individual methane-oxidizing aggregates is currently lacking. Here, we use the microanalytical method FISH-SIMS (fluorescence in situ hybridization-secondary ion mass spectrometry) to describe the physiological traits and anabolic activity of individual methanotrophic consortia, specifically tracking ^(15)N-labelled protein synthesis to examine the effects of organization and size on the metabolic activity of the syntrophic partners. Patterns of 15N distribution within individual aggregates showed enhanced ^(15)N assimilation in ANME-2 cells relative to the co-associated DSS revealing a decoupling in anabolic activity between the partners. Protein synthesis in ANME-2 cells was sustained throughout the core of individual ANME-2/DSS consortia ranging in size range from 4 to 20 μm. This indicates that metabolic activity of the methane-oxidizing archaea is not limited to, or noticeably enhanced at the ANME−2/DSS boundary. Overall, the metabolic activity of both syntrophic partners within consortia was greater than activity measured in representatives of the ANME-2 and DSS observed alone, with smaller ANME-2/DSS aggregates displaying a tendency for greater ^(15)N uptake and doubling times ranging from 3 to 5 months. The combination of ^(15)N-labelling and FISH-SIMS provides an important perspective on the extent of heterogeneity within methanotrophic aggregates and may aid in constraining predictive models of activity and growth by these syntrophic consortia.

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Orphan, Victoria J.0000-0002-5374-6178
House, Christopher H.0000-0002-4926-4985
Additional Information:Journal compilation © 2009 Society for Applied Microbiology and Blackwell Publishing Ltd. Received 10 October, 2008; accepted 6 February, 2009. We thank A. Schmitt, K. McKeegan and G. Jarzebinski from the UCLA ion microprobe facility, and C. Gammon, A. Pernthaler and T. Embaye for their technical assistance with this project. We also acknowledge S. Goffredi, D. Fike, K.-U. Hinrichs and two anonymous reviewers for critical reading of this manuscript and thank S. Joye and A. Pearson for fruitful discussions. We are grateful to the captain, pilots, crew and shipboard research parties of the R/V Western Flyer and R/V Atlantis (AT 15-11) for their invaluable support. This work was supported by grants from the National Science Foundation (MCB-0348492), NOAA Undersea Research Program (UAF 05-0132), and the Gordon and Betty Moore Foundation (to V.J.O.). A.M.G. was supported by a NSF Graduate Research Fellowship. Appendix S1. Heterogeneity in δ13C and 15N assimilation in sediment incubations exhibiting low anaerobic oxidation of methane activity. Fig. S1. Relationship between δ13C (‰) and 15N (atom %) for shell ANME-2/DSS after a 5-day incubation with 15N-labelled ammonium or amino acids from (A) PC-55 (diamonds; n = 6) and PC-76 (squares; n = 10). ‘z’ symbol denotes paired δ13C/15N values for a mixed ANME-2/DSS aggregate. (B) Paired δ13C/15N values for PC-59 cell aggregates after 4-day incubation (open symbols) and 85-day incubation (closed symbols) with ammonium and amino acids. Shell aggregates are represented by a square symbol; mixed aggregates (triangle) and mono-specific ANME-2 clusters are represented by a circle. Plus signs denote ANME/DSS shell aggregates from control incubation without exogenous 15N-labelled nitrogen. In both panels, plotted values include heaviest δ13C (red colour) and lightest δ13C (black) data points for each ANME-2 or ANME/DSS aggregate measured during the FISH-SIMS analysis. Fig. S2. Comparison of the relationship between aggregate size and enrichment in 15Nbiomass (atom %) for shell consortia. A. Maximum 15N enrichment after 5-day incubation for shell aggregates in PC-76 and PC-55. B. Box plot showing the average 15N value, range, and 95% confidence intervals for shell aggregates with diameters between 2 and 7 µm (mean 15N atom % = 1.8, n = 9) and aggregates ranging between 7 and 20 µm (mean 15N atom % = 1.4, n = 6). C. Maximum 15N enrichment after 112-day incubation for shell aggregates in PC-76. Although a general trend of greater 15N assimilation by smaller shell aggregates was present, the statistical significance of aggregate size and 15N enrichment was not observed (P = 0.44, Wilcoxon test). Table S1. Fluorescence in situ hybridization (FISH) quantification of per cent change in ANME and bacteria over time. Table S2. Temporal variation in ratio of ANME-2/DSS shell aggregate size in PC-76.
Funding AgencyGrant Number
National Oceanic and Atmospheric Administration (NOAA)UAF 05-0132
Gordon and Betty Moore FoundationUNSPECIFIED
Issue or Number:7
Record Number:CaltechAUTHORS:20090909-123033461
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:15698
Deposited By: George Porter
Deposited On:30 Sep 2009 19:50
Last Modified:09 Mar 2020 13:18

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