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METHODS: Fluxes were measured using a system of 8 automatic gas-sampling chambers made of transparent Lexan (n=3 each in the palsa and bog habitats, and n=2 in the fen habitat). Chambers were initially installed in the three habitat types at Stordalen Mire in 2001 (Bäckstrand et al., 2008) and the chamber lids were replaced in 2011 with the current design, similar to that described by Bubier et al 2003. Chambers cover an area of 0.2 m2 (45 cm x 45 cm), with a height ranging from 15-75 cm depending on habitat vegetation. At the Palsa and bog site the chamber base is flush with the ground and the chamber lid (15 cm in height) lifts clear of the base between closures. At the fen site the chamber base is raised 50–60 cm on lexon skirts to accommodate large stature vegetation. The chambers are connected to the gas analysis system, located in an adjacent temperature-controlled cabin, by 3/8” Dekoron tubing through which air is circulated at approximately 2.5 L min-1. Each chamber lid is closed once every 3 hours for a period of 8 min, with a 5 min flush period before and after lid closure. Gas concentration in the chamber headspace was measured with a Los Gatos Research (LGR) Fast Greenhouse Gas Analyzer, with timing control and data acquisition using a Campbell CR10x (Holmes et al., 2022). References: Bäckstrand, K., Crill, P. M., Mastepanov, M., Christensen, T. R. & Bastviken, D. Total hydrocarbon flux dynamics at a subarctic mire in northern Sweden. Journal of Geophysical Research 113, (2008). Bubier, J. L., Crill, P. M., Mosedale, A., Frolking, S. & Linder, E. Peatland responses to varying interannual moisture conditions as measured by automatic CO2 chambers. Global Biogeochemical Cycles 17, (2003). Holmes, M. E., Crill, P. M., Burnett, W. C., McCalley, C. K., Wilson, R. M., Frolking, S., Chang, K. ‐Y., Riley, W. J., Varner, R. K., Hodgkins, S. B., IsoGenie Project Coordinators, IsoGenie Field Team, McNichol, A. P., Saleska, S. R., Rich, V. I., Chanton, J. P. (2022). Carbon accumulation, flux, and fate in Stordalen Mire, a permafrost peatland in transition. Global Biogeochemical Cycles, 36, e2021GB007113, doi:10.1029/2021GB007113. McCalley, C.K., B.J. Woodcroft, S.B. Hodgkins, R.A. Wehr, E-H. Kim, R. Mondav, P.M. Crill, J.P. Chanton, V.I. Rich, G.W. Tyson, S.R. Saleska (2014), Methane dynamics regulated by microbial community response to permafrost thaw, Nature, 514:478-481, doi:10.1038/nature13798. FUNDING: This research is a contribution of the EMERGE Biology Integration Institute, funded by the National Science Foundation, Biology Integration Institutes Program, Award # 2022070.We thank the Swedish Polar Research Secretariat and SITES for the support of the work done at the Abisko Scientific Research Station. SITES is supported by the Swedish Research Council's grant 4.3-2021-00164.This study was also funded by the Genomic Science Program of the United States Department of Energy Office of Biological and Environmental Research, grant #s DE-SC0004632, DE-SC0010580, and DE-SC0016440.These autochamber measurements were also supported by a grant from the US National Science Foundation MacroSystems program (NSF EF 1241037, PI Varner).more » « less
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Autochamber-based CH4 fluxes and δ13C values measured with a Tunable Infrared Laser Direct Absorption Spectrometer (TILDAS, Aerodyne Research Inc.); and ancillary data, including CO2 fluxes (measured with a LGR Greenhouse Gas Analyzer), temperatures, atmospheric pressure, and photosynthetically active radiation (PAR). In addition to the data published here, data from 2011 is also available in the supplementary files to McCalley et al. (2014) under the Source data to Fig. 1 link. METHODS: Methane fluxes were measured using a system of 8 automatic gas-sampling chambers made of transparent Lexan (n=3 each in the palsa and bog habitats, and n=2 in the fen habitat). Chambers were initially installed in the three habitat types at Stordalen Mire in 2001 (Bäckstrand et al., 2008) and the chamber lids were replaced in 2011 with the current design, similar to that described by Bubier et al 2003. Chambers cover an area of 0.2 m2 (45 cm x 45 cm), with a height ranging from 15-75 cm depending on habitat vegetation. At the Palsa and bog site the chamber base is flush with the ground and the chamber lid (15 cm in height) lifts clear of the base between closures. At the fen site the chamber base is raised 50–60 cm on lexon skirts to accommodate large stature vegetation. The chambers are instrumented with thermocouples measuring air and surface ground temperature, and water table depth and thaw depth are measured manually 3–5 times per week. The chambers are connected to the gas analysis system, located in an adjacent temperature-controlled cabin, by 3/8” Dekoron tubing through which air is circulated at approximately 2.5 L min-1. Each chamber lid is closed once every 3 hours for a period of 8 min, with a 5 min flush period before and after lid closure. We measured methane concentration using a Tunable Infrared Laser Direct Absorption Spectrometers (TILDAS, Aerodyne Research Inc.) connected to the main chamber circulation using ¼” Dekoron tubing (McCalley et al 2014). Calibrations were done every 90 min using 3 calibration gases spanning the observed concentration range (1.8–10 ppm). For each autochamber closure we calculated flux using a method consistent with that detailed by Bäckstrand et al 2008 for CO2 and total hydrocarbons, using a linear regression of changing headspace CH4 concentration over a period of 2.5 min. Eight 2.5 min regressions were calculated, staggered by 15 sec, and the most linear fit (highest r2) was then used to calculate flux. Daily average flux for each chamber was used to calculate daily flux and standard error for each cover type. References: Bäckstrand, K., Crill, P. M., Mastepanov, M., Christensen, T. R. & Bastviken, D. Total hydrocarbon flux dynamics at a subarctic mire in northern Sweden. Journal of Geophysical Research 113, (2008). Bubier, J. L., Crill, P. M., Mosedale, A., Frolking, S. & Linder, E. Peatland responses to varying interannual moisture conditions as measured by automatic CO2 chambers. Global Biogeochemical Cycles 17, (2003). McCalley, C.K., B.J. Woodcroft, S.B. Hodgkins, R.A. Wehr, E-H. Kim, R. Mondav, P.M. Crill, J.P. Chanton, V.I. Rich, G.W. Tyson, S.R. Saleska (2014), Methane dynamics regulated by microbial community response to permafrost thaw, Nature, 514:478-481, doi:10.1038/nature13798. FILES: Files are named with the year or date range, followed by a suffix indicating data resolution: *_CH4output_clean_ckm.txt - Individual measurements of CH4 fluxes (CH4Flux), CO2 fluxes (CO2flux; for select years), and δ13C signature of emitted CH4 (Flux13CH4) for each chamber closure. CH4FluxRsq is the R2 value of the linear fit used to calculate CH4 flux, CO2Rsq is the R2 value of the linear fit used to calculate CO2 flux, and Flux13CH4_stdev is the standard deviation of the δ13C signature (standard deviation of the intercept of the Keeling plot). *_DailyCH4output_ckm.txt - Daily average CH4 fluxes (CH4Flux) and δ13C values (13CH4), grouped by site: Palsa, Bog, Fen, and Chamber 9 (bog/fen transition); along with standard deviations (stdev) and standard errors (se) of the flux or δ13C for each site type. For the Palsa, Bog, and Fen sites, these averages are calculated by chamber (n=3 for Palsa and Bog, n=2 for Fen), so each chamber's daily average is calculated, and then a daily average for that site is calculated as the average of the chambers. For Chamber 9 (bog/fen intermediate; n=1 chamber), averages are calculated by day as there are no chamber replicates. MEASUREMENT UNITS (same for both file types): CH4 flux: mg CH4 m−2 hr−1 CO2 flux: mg C m−2 h−1 δ13C: ‰ Temperature: °C Air pressure: mbar PAR: µmol photons m−2 s−1 FUNDING: This research is a contribution of the EMERGE Biology Integration Institute, funded by the National Science Foundation, Biology Integration Institutes Program, Award # 2022070.We thank the Swedish Polar Research Secretariat and SITES for the support of the work done at the Abisko Scientific Research Station. SITES is supported by the Swedish Research Council's grant 4.3-2021-00164.This study was also funded by the Genomic Science Program of the United States Department of Energy Office of Biological and Environmental Research, grant #s DE-SC0004632, DE-SC0010580, and DE-SC0016440.Autochamber measurements between 2013 and 2017 were supported by a grant from the US National Science Foundation MacroSystems program (NSF EF 1241037, PI Varner).more » « less
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Methane (CH4) emissions in Stordalen Mire (northern Sweden), estimated via two different approaches: "Paint by number" (field ch4_modified_prj.tif): CH4 emission across the landscape calculated via “paint-by-number” approach, using 2014 autochamber-based flux measurements (https://doi.org/10.5281/zenodo.14052690) mapped to landcover classifications (https://doi.org/10.5281/zenodo.15042233). DNDC-modeled (Modeled CH4.tif): CH4 emission across the landscape modeled via Wetland-DNDC (https://www.dndc.sr.unh.edu/), driven by remotely sensed landcover classifications (https://doi.org/10.5281/zenodo.15042233), water table depth (https://doi.org/10.5281/zenodo.15092752), climate data (provided by the Abisko Scientific Research Station), and soil parameters (defined as in Deng et al. 2014, 2017). The DNDC model was run on vegetation and water table clusters (determined by k-means clustering), and model output was spatially assigned to each map pixel. Modeled CH4 emissions account for CH4 production from DOC (Randomforest_stack_epsg32634_extent_kmeansclass10_CH4 prod from DOC.tif) and from CO2 (Randomforest_stack_epsg32634_extent_kmeansclass10_CH4 prod from CO2.tif), minus oxidation (Randomforest_stack_epsg32634_extent_kmeansclass10_CH4 oxid.tif). The model also outputs a map of CH4 isotopic composition (δ13C-CH4) of emissions (Randomforest_stack_epsg32634_extent_kmeansclass10_Delta CH4 flux.tif). The difference between these approaches is provided as a difference map (CH4diff.tif), calculated as the "paint-by-number" (PBN) emissions (field ch4_modified_prj.tif) minus the Wetland-DNDC modeled emissions (Modeled CH4.tif). These images are GeoTIFFs with embedded georeferencing information. FUNDING: National Aeronautics and Space Administration, Interdisciplinary Science program: From Archaea to the Atmosphere (award # NNX17AK10G). National Science Foundation, Biology Integration Institutes Program: EMERGE Biology Integration Institute (award # 2022070). United States Department of Energy Office of Biological and Environmental Research, Genomic Science Program: The IsoGenie Project (grant #s DE-SC0004632, DE-SC0010580, and DE-SC0016440). National Science Foundation, MacroSystems program (grant # EF-1241037). We thank the Swedish Polar Research Secretariat and SITES for the support of the work done at the Abisko Scientific Research Station. SITES is supported by the Swedish Research Council's grant 4.3-2021-00164.more » « less
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Abstract The dynamics of methane (CH4) cycling in high-latitude peatlands through different pathways of methanogenesis and methanotrophy are still poorly understood due to the spatiotemporal complexity of microbial activities and biogeochemical processes. Additionally, long-termin situmeasurements within soil columns are limited and associated with large uncertainties in microbial substrates (e.g. dissolved organic carbon, acetate, hydrogen). To better understand CH4cycling dynamics, we first applied an advanced biogeochemical model,ecosys, to explicitly simulate methanogenesis, methanotrophy, and CH4transport in a high-latitude fen (within the Stordalen Mire, northern Sweden). Next, to explore the vertical heterogeneity in CH4cycling, we applied the PCMCI/PCMCI+ causal detection framework with a bootstrap aggregation method to the modeling results, characterizing causal relationships among regulating factors (e.g. temperature, microbial biomass, soil substrate concentrations) through acetoclastic methanogenesis, hydrogenotrophic methanogenesis, and methanotrophy, across three depth intervals (0–10 cm, 10–20 cm, 20–30 cm). Our results indicate that temperature, microbial biomass, and methanogenesis and methanotrophy substrates exhibit significant vertical variations within the soil column. Soil temperature demonstrates strong causal relationships with both biomass and substrate concentrations at the shallower depth (0–10 cm), while these causal relationships decrease significantly at the deeper depth within the two methanogenesis pathways. In contrast, soil substrate concentrations show significantly greater causal relationships with depth, suggesting the substantial influence of substrates on CH4cycling. CH4production is found to peak in August, while CH4oxidation peaks predominantly in October, showing a lag response between production and oxidation. Overall, this research provides important insights into the causal mechanisms modulating CH4cycling across different depths, which will improve carbon cycling predictions, and guide the future field measurement strategies.more » « lessFree, publicly-accessible full text available February 11, 2026
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Abstract Quantifying the temperature sensitivity of methane (CH4) production is crucial for predicting how wetland ecosystems will respond to climate warming. Typically, the temperature sensitivity (often quantified as a Q10value) is derived from laboratory incubation studies and then used in biogeochemical models. However, studies report wide variation in incubation-inferred Q10values, with a large portion of this variation remaining unexplained. Here we applied observations in a thawing permafrost peatland (Stordalen Mire) and a well-tested process-rich model (ecosys) to interpret incubation observations and investigate controls on inferred CH4production temperature sensitivity. We developed a field-storage-incubation modeling approach to mimic the full incubation sequence, including field sampling at a particular time in the growing season, refrigerated storage, and laboratory incubation, followed by model evaluation. We found that CH4production rates during incubation are regulated by substrate availability and active microbial biomass of key microbial functional groups, which are affected by soil storage duration and temperature. Seasonal variation in substrate availability and active microbial biomass of key microbial functional groups led to strong time-of-sampling impacts on CH4production. CH4production is higher with less perturbation post-sampling, i.e. shorter storage duration and lower storage temperature. We found a wide range of inferred Q10values (1.2–3.5), which we attribute to incubation temperatures, incubation duration, storage duration, and sampling time. We also show that Q10values of CH4production are controlled by interacting biological, biochemical, and physical processes, which cause the inferred Q10values to differ substantially from those of the component processes. Terrestrial ecosystem models that use a constant Q10value to represent temperature responses may therefore predict biased soil carbon cycling under future climate scenarios.more » « less
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Abstract Climate change is disproportionately warming northern peatlands, which may release large carbon stores via increased microbial activity. While there are many unknowns about such microbial responses, virus roles are especially poorly characterized with studies to date largely restricted to “bycatch” from bulk metagenomes. Here, we used optimized viral particle purification techniques on 20 samples along a highly contextualized peatland permafrost thaw gradient, extracted and sequenced viral particle DNA using two library kits to capture single-stranded (ssDNA) and double-stranded (dsDNA) virus genomes (40 total viromes), and explored their diversity and potential ecosystem impacts. Both kits recovered similar dsDNA virus numbers, but only one also captured thousands of ssDNA viruses. Combining these data, we explored population-level ecology using genomic representation from 9,560 viral operational taxonomic units (vOTUs); nearly a 4-fold expansion from permafrost-associated soils, and 97% of which were novel when compared against large datasets from soils, oceans, and the human gut.In silicopredictions identified putative hosts for 44% (4,149 dsDNA + 17 ssDNA) of the identified vOTUs spanning 2 eukaryotic, 12 archaeal, and 30 bacterial phyla. The recovered vOTUs encoded 1,684 putative auxiliary metabolic genes (AMGs) and other metabolic genes carried by ∼10% of detected vOTUs, of which 46% were related to carbon processing and 644 were novel. These AMGs grouped into five functional categories and 11 subcategories, and nearly half (47%) of the AMGs were involved in carbon utilization. Of these, 112 vOTUs encoded 123 glycoside hydrolases spanning 15 types involved in the degradation of polysaccharides (e.g., cellulose) to monosaccharides (e.g., galactose), or further monosaccharide degradation, which suggests virus involvement in myriad metabolisms including fermentation and central carbon metabolism. These findings expand the scope of viral roles in microbial carbon processing and suggest viruses may be critical for understanding the fate of soil organic carbon in peatlands.more » « less
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