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Kinetic and temperature sensitivity properties of soil exoenzymes through the soil profile down to one-meter depth at a temperate coniferous forest (Blodgett, CA)

This dataset contains data on kinetic and temperature sensitivity parameters of the exoenzymes β-glucosidase (BG), leucine/leucyl aminopeptidase (LAP) and acid phosphatase (AP) across a soil profile down to 90 cm depth at Blodgett forest, CA, USA. These data were generated to determine if kinetic and thermal properties of microbial exoenzymes involved in organic matter decomposition varied with soil depth, following variation in soil properties and microbial communities, as part of the study: Alves et al. (2021). Kinetic Properties of Microbial Exoenzymes Vary with Soil Depth but Have Similar Temperature Sensitivities Through the Soil Profile. Frontiers in Microbiology 12:3618. https://doi.org/10.3389/fmicb.2021.735282. This research was performed within the framework of the TES Belowground Biogeochemistry SFA project, in particular association with a long-term field warming experiment of the whole soil profile at Blodgett forest. Samples for this work were collected from locations representative of the field experimental plots. Potential enzyme activity rates were measured using laboratory fluorometric assays with soils collected at 0-10, 10-20, 30-40, 50-60, 60-70 and 80-90 cm deep in biological triplicates (i.e., three soil cores collected at different representative locations). Assays with each soil were conducted over a gradient of eight substrate concentrations per enzyme, and incubated at 4, 10, 16, 25, 35 or 50°C. Enzyme Michaelis-Menten kinetics were modeled over the eight substrate concentrations at each temperature, and the temperature sensitivity of the kinetic parameters was modeled over the six temperatures using linear Arrhenius/Q10 and non-linear Macromolecular Rate Theory (MMRT) models. The dataset includes the fully processed enzyme activity rate data used to model Michaelis-Menten kinetics, calculated kinetic and temperature sensitivity parameters, and basic soil and microbial biomass chemistry for each sample. All data is provided for each individual biological replicate, and kinetic and temperature sensitivity parameters are also provided as means of the biological replicates. The dataset also includes all raw measurement data and code used to parse, combine and perform the analyses described by Alves et al. (2021). For file descriptions, see the file-level metadata files: “enzymes_dataProcessed_flmd.xlsx” (processed data); “enzymes_dataRaw_flmd.xlsx” (compressed raw data and metadata); and “enzymes_code_flmd.xlsx” (compressed code). The experimental design, list of parameters measured, soil and microbial biomass chemistry data, and means of biological replicates for kinetic and temperature sensitivity parameters are also provided as human-readable tables in file “enzymeTraits_design_results_tables.xlsx”.

54 ENVIRONMENTAL SCIENCES↗

Soil Oxygen Limits Microbial Phosphorus Utilization in Humid Tropical Forest Soils

Soil phosphorus (P) availability is of special interest in many humid tropical forests, especially those on highly weathered, iron (Fe)- and aluminum (Al)-rich soils where P often limits net primary productivity. Phosphorus cycling is partly dependent on the ability of microbes to compete for P with Fe and Al minerals, which strongly bind P. Soil P availability is also indirectly affected by soil redox conditions due to its effects on microbial activity and reductive dissolution of Fe oxides that may weaken Fe-O-P sorption strength. Here, we explored P sorption, soil Fe (II) concentrations, soil CO2 production, organic and inorganic P pools, and microbial biomass P in tropical soils that typically experience frequent low redox (valley soils), or fluctuating redox conditions (slope soils). Soils from both topographic positions were pre-incubated under oxic or anoxic headspaces and then amended with a mixture of P (as orthophosphate) and carbon (C, as acetate, to maintain microbial activity) and incubated in the dark for 24 h. Phosphorus sorption to the mineral phase occurred on a time scale of seconds to minutes in valley and slope soils, reflecting strong abiotic P sorption capacity. Valley soils were characterized by inherently higher Fe(II) concentrations and lower respiration rates. Under anoxic headspaces, Fe(II) concentrations increased 3-to 5-fold in the both soils. Soil respiration and microbial P utilization declined significantly in both soils under anoxic conditions, regardless of Fe(II) concentrations. Microbial P concentrations were highest when slope soils were incubated under an oxic headspace, despite the high P sorption under these conditions. Our results suggest that microbial P utilization is indirectly limited by low O2 availability and that microbes are able to effectively compete with minerals for P under Fe-oxidizing conditions. These results emphasize the central role of soil microorganisms in regulating P availability, even in the presence of strong abiotic sorption capacity.

54 ENVIRONMENTAL SCIENCES↗

Sandy Soil Microaggregates: Rethinking Our Understanding of Hydraulic Function

Core Ideas Sandy soil microaggregates are comprised of a solid sand core encrusted with fines. Thick films control a significant proportion of water retention in coarse sandy soil. Three water pools drain sequentially: capillary water, thick films, and adsorptive films. Film seepage rates should be considered when irrigating sandy soil. This study investigated the peculiar structure of microaggregates in coarse sandy soils that exhibit only external porosity and investigated their control on soil hydrology. The microstructure underpins a hydrologic existence that differs from finer textured soils where aggregates have internal porosity. Understanding the impact of these microaggregates on soil hydrology will permit improved agricultural irrigation management and estimates associated with ecosystem capacity and resiliency. Microstructure was investigated using a digital microscope, and aspects of the structure were quantified by sedimentation and computed microtomography. Sandy soil microaggregates were observed to be comprised of a solid sand‐grain core that is coated with fines, presumably cemented by organic media. This microstructure leads to three distinct water pools during drainage: capillary water, followed by thick films (1–20 μm) enveloping the outer surfaces of the crusted microaggregates, followed by adsorbed thin films (<1 μm). The characteristics of the thick films were investigated using an analytical model. These films may provide as much as 10 to 40% saturation in the range of plant‐available water. Using lubrication theory, it was predicted that thick film drainage follows a power law function with an exponent of 2. Thick films may also have a role in the geochemical evolution of soils and in ecosystem function because they provide contiguous water and gas phases at relatively high moisture contents. And, because the rough outer crust of these microaggregates can provide good niches for microbial activity, biofilm physics will dominate thick film processes, and consequently hydrologic, biologic, and geochemical functions for coarse sandy soils.

Paradiś, Ashley↗

Contributions of anoxic microsites to soil carbon protection across soil textures

Anoxic microsites, zones of oxygen depletion in otherwise oxic soils, may slow soil C turnover. However, the abundance of anoxic microsites and their contribution to soil C protection is yet undefined. In this study, we determine the contribution of anoxic microsites to soil C protection in soils of three distinct textures (clay loam, loam, sandy loam) across a range of soil moistures. We examined the influence of soil oxygen supply by increasing oxygen content in the incubation atmosphere (oxygen enrichment) and through disaggregation. We attributed increases in CO 2 efflux to the aeration of anoxic microsites. The contribution of anoxic microsites to soil C protection increased with decreasing clay content. Clay loam CO 2 efflux was relatively unaffected by aeration. Moderately moist, loam soils had CO 2 effluxes that did not increase with oxygen enrichment but increased by 375% upon disaggregation. Sandy loam soil CO 2 efflux increased by 50–75% with oxygen enrichment and 250% with disaggregation. Geochemical and microbial data reveal that anoxic microsite abundance also increased with decreasing clay content. The proportion of acid extractable Fe present as Fe(II) increased with decreasing clay content, and methanogens were more abundant in loam and sandy loam soils. Our results suggest that oxygen demand, rather than supply, can regulate anoxic microsite formation and that anoxic protection of soil C can be diminished through physical disruption of soil structure. Our findings further illustrate that anoxic microsites should be included in conceptual models of soil C protection to avoid soil C loss and improve predictions of soil C response to disturbance.

58 GEOSCIENCES↗

Soil characteristics and land-use drive bacterial community assembly patterns

ABSTRACT Land-use and soil characteristics drive variation in soil community composition, but the influences of these factors on dispersal and community assembly at regional scale remain poorly characterized. Land-use remains a consistent driver of soil community composition even when exhibiting patchy spatial distribution at regional scale. In addition, disturbed and early successional soils often exhibit stochastic community assembly patterns. These observations suggest local community composition is influenced by dispersal and assembly from regional species pools. We examined bacterial community assembly within agricultural cropland, old-field, and forested sites across 10 landscapes in the region around Ithaca, New York (USA). We found that the Sloan neutral model explained assembly well at regional scale (R2 = 0.763), but that both soil pH and land-use imposed selection that shaped community composition. We show that homogeneous selection was a dominant assembly process with respect to both soil pH and land-use regime, but that these two factors interacted in their effects on bacterial community assembly. We conclude that bacterial community assembly at a regional scale is driven by dispersal from regional species pools and local selection on the basis of soil pH and other soil characteristics that vary with land-use.

Microbiology↗

Does elevated atmospheric CO 2 affect soil carbon burial and soil weathering in a forest ecosystem?

Most experimental studies measuring the effects of climate change on terrestrial C cycling have focused on processes that occur at relatively short time scales (up to a few years). However, climate-soil C interactions are influenced over much longer time scales by bioturbation and soil weathering affecting soil fertility, ecosystem productivity, and C storage. Elevated CO 2 can increase belowground C inputs and stimulate soil biota, potentially affecting bioturbation, and can decrease soil pH which could accelerate soil weathering rates. To determine whether we could resolve any changes in bioturbation or C storage, we investigated soil profiles collected from ambient and elevated-CO 2 plots at the Free-Air Carbon-Dioxide Enrichment (FACE) forest site at Oak Ridge National Laboratory after 11 years of 13 C-depleted CO 2 release. Profiles of organic carbon concentration, δ 13 C values, and activities of 137 Cs, 210 Pb, and 226 Ra were measured to ~30 cm depth in replicated soil cores to evaluate the effects of elevated CO 2 on these parameters. Bioturbation models based on fitting advection-diffusion equations to 137 Cs and 210 Pb profiles showed that ambient and elevated-CO 2 plots had indistinguishable ranges of apparent biodiffusion constants, advection rates, and soil mixing times, although apparent biodiffusion constants and advection rates were larger for 137 Cs than for 210 Pb as is generally observed in soils. Temporal changes in profiles of δ 13 C values of soil organic carbon (SOC) suggest that addition of new SOC at depth was occurring at a faster rate than that implied by the net advection term of the bioturbation model. Ratios of ( 210 Pb/ 226 Ra) may indicate apparent soil mixing cells that are consistent with biological mechanisms, possibly earthworms and root proliferation, driving C addition and the mixing of soil between ~4 cm and ~18 cm depth. Lastly, burial of SOC by soil mixing processes could substantially increase the net long-term storage of soil C and should be incorporated in soil-atmosphere interaction models.

54 ENVIRONMENTAL SCIENCES↗

Soil carbon sequestration and land use change associated with biofuel production: Empirical evidence

Soil organic carbon (SOC) change can be a major impact of land use change (LUC) associated with biofuel feedstock production. By collecting and analyzing data from worldwide field observations with major LUCs from cropland, grassland and forest to lands producing biofuel crops (i.e., corn, switchgrass, Miscanthus, poplar and willow), we were able to estimate SOC response ratios and sequestration rates and evaluate the effects of soil depth and time scale on SOC change. Both the amount and rate of SOC change were highly dependent on the specific land transition. Irrespective of soil depth or time horizon, cropland conversions resulted in an overall SOC gain of 6-14% relative to initial SOC level, while conversion from grassland or forest to corn (without residue removal) or poplar caused significant carbon loss (9-35%). No significant SOC changes were observed in land converted from grasslands or forests to switchgrass, Miscanthus or willow. The SOC response ratios were similar in both 0-30 and 0-100 cm soil depths in most cases, suggesting SOC changes in deep soil and that use of top soil only for SOC accounting in biofuel life cycle analysis (LCA) might underestimate total SOC changes. Soil carbon sequestration rates varied greatly among studies and land transition types. Generally, the rates of SOC change tended to be the greatest during the 10 years following land conversion, and had declined to approach 0 within about 20 years for most LUCs. Observed trends in SOC change were generally consistent with previous reports. Soil depth and duration of study significantly influence SOC change rates and so should be considered in carbon emission accounting in biofuel LCA. High uncertainty remains for many perennial systems, field trials and modeling efforts are needed to determine the site- and system-specific rates and direction of change associated with their production.

corn↗

Comparing vis–NIRS, LIBS, and Combined vis–NIRS‐LIBS for Intact Soil Core Soil Carbon Measurement

Core Ideas First rigorous integration of VisNIR and LIBS spectroscopic techniques for soil profile C measurement. Field‐moist intact soil cores scanned with LIBS and VisNIR simulating in situ measurements. Combined VisNIR‐LIBS wavelengths selected by MRCE were split nearly equally between sensors, suggesting molecular and elemental information was successfully integrated for predicting TC, IC, and SOC. Stoichiometric relationships in LIBS elemental emissions are more strongly correlated to IC concentration than were carbonate absorptions in VisNIR. Lack of variation and consistent, definable chemical composition and stoichiometry likely contributed to poor LIBS and combined VisNIR – LIBS prediction accuracy for SOC. Soil organic carbon (SOC) measurement is critically important to quantify regional and global soil C stocks and better understand soil C biogeochemical processes. Recent studies employing laser‐induced breakdown spectroscopy (LIBS) and visible‐near infrared diffuse reflectance spectroscopy (vis–NIRS) indicate their potential for in situ soil C determination. Visible and near infrared diffuse reflectance spectroscopy and LIBS spectroscopy fundamentally differ and we hypothesize that their integration would provide improved soil C predictions. We report the first rigorous integration of vis–NIRS and LIBS, evaluating the precision of vis–NIRS, LIBS, and combined vis–NIRS‐LIBS spectra for simulated in situ soil profile total C (TC), inorganic C (IC) and SOC measurement. Three multivariate variable selection and regression approaches were evaluated for soil C prediction. The highest soil C prediction accuracies were observed using multivariate regression with covariance estimation (MRCE). Inorganic C was best predicted by LIBS, vis–NIRS provided better SOC predictions, and TC was best predicted using combined vis–NIRS‐LIBS data. Combined vis–NIRS‐LIBS did not consistently increase soil C prediction accuracy. Soil organic C was not well predicted, presumably due to challenges associated with scanning surfaces of intact soil cores, variable SOC chemistries, and low SOC variation in the dataset. Considering the challenging conditions under which combined vis–NIRS– LIBS was tested for soil C measurement, data integration and model calibrations had acceptable performance. Further testing under more controlled soil conditions with samples containing greater SOC diversity is necessary to determine the technical potential of combined vis–NIRS/LIBS for soil C determination.

Bricklemyer, Ross S.↗

Patterns of longer-term climate change effects on CO 2 efflux from biocrusted soils differ from those observed in the short term

Biological soil crusts (biocrusts) are predicted to be sensitive to the increased temperature and altered precipitation associated with climate change. We assessed the effects of these factors on soil carbon dioxide (CO 2 ) balance in biocrusted soils using a sequence of manipulations over a 9-year period. We warmed biocrusted soils by 2 and, later, by 4 °C to better capture updated forecasts of future temperature at a site on the Colorado Plateau, USA. We also watered soils to alter monsoon-season precipitation amount and frequency and had plots that received both warming and altered precipitation treatments. Within treatment plots, we used 20 automated flux chambers to monitor net soil exchange (NSE) of CO 2 hourly, first in 2006–2007 and then again in 2013–2014, for a total of 39 months. Net CO 2 efflux from biocrusted soils in the warming treatment increased a year after the experiment began (2006–2007). However, after 9 years and even greater warming (4 °C), results were more mixed, with a reversal of the increase in 2013 (i.e., controls showed higher net CO 2 efflux than treatment plots) and with similarly high rates in all treatments during 2014, a wet year. Over the longer term, we saw evidence of reduced photosynthetic capacity of the biocrusts in response to both the temperature and altered precipitation treatments. Patterns in biocrusted soil CO 2 exchange under experimentally altered climate suggest that (1) warming stimulation of CO 2 efflux was diminished later in the experiment, even in the face of greater warming; and (2) treatment effects on CO 2 flux patterns were likely driven by changes in biocrust species composition and by changes in root respiration due to vascular plant responses.

54 ENVIRONMENTAL SCIENCES↗

Global patterns and controls of soil organic carbon dynamics as simulated by multiple terrestrial biosphere models. Current status and future directions

Soil is the largest organic carbon (C) pool of terrestrial ecosystems, and loss from soil accounts for a large pro portion of land-atmosphere C exchange. Due to large pool size and variable residence time from years to millennia, even small changes in soil organic C(SOC) have substantial effects on the terrestrial C budget, thereby affecting atmospheric carbon dioxide (CO2)concentration and climate change. In the past decades, a wide variety of studies have been conducted to quantify global SOC stocks and soil exchange with the atmosphere through site measurements, inventories, and empirical/process-based modeling. However, these estimates are highly uncertain and identifying major driving forces controlling soil C storage and fluxes remains a key research challenge his study has compiled century-long (1901-2010)estimates of SOC storage and heterotrophic respiration (Rh) from ten terrestrial biosphere models (TBMs) in the Multi-scale Synthesis and Terrestrial Model Intercomparison Project (MsTMIP) and two observation based datasets. The ten-TBM ensemble shows that global SOC estimate range from 4 to 2111 Pg C (1 Pg = 10 15 g) with a median value of 1158 Pg C33 in 2010. Modeling approach estimates a broad range of Rh from 35 to 69 Pg C yr -1 with a median value of 51Pg C yr -1 during 200–2010. The largest uncertainty in SOC stocks exists in the 40–65°N latitude band while Rh differences are the largest in the tropics. All the models agreed that climate and land use changes have decreased SOC stocks while elevated CO 2 and atmospheric nitrogen deposition have increased SOC stocks though the response varied significantly among models. Model representations of temperature and moisture sensitivity,nutrient limitation and land use partially explain the divergent estimates of global SOC stocks and soil fluxes in this study. In addition, major sources of uncertainty from model estimation include exclusion of SOC storage in wetlands and peatlands as well as C storage in deep soil layers.

54 ENVIRONMENTAL SCIENCES↗

Arsenic hyperaccumulator Pteris vittata shows reduced biomass in soils with high arsenic and low nutrient availability, leading to increased arsenic leaching from soil

Plant-soil interactions affect arsenic and nutrient availability in arsenic-contaminated soils, with implications for arsenic uptake and tolerance in plants, and leaching from soil. In 22-week column experiments, we grew the arsenic hyperaccumulating fern Pteris vittata in a coarse- and a medium-textured soil to determine the effects of phosphorus fertilization and mycorrhizal fungi inoculation on P. vittata arsenic uptake and arsenic leaching. We investigated soil arsenic speciation using synchrotron-based spectromicroscopy. Greater soil arsenic availability and lower nutrient content in the coarse-textured soil were associated with greater fern arsenic uptake, lower biomass (apparently a metabolic cost of tolerance), and arsenic leaching from soil, due to lower transpiration. P. vittata hyperaccumulated arsenic from coarse- but not medium-textured soil. Mass of plant-accumulated arsenic was 1.2 to 2.4 times greater, but aboveground biomass was 74% smaller, in ferns growing in coarse-textured soil. In the presence of ferns, mean arsenic loss by leaching was 195% greater from coarse- compared to the medium-textured soil, and lower across both soils compared to the absence of ferns. In the medium-textured soil arsenic concentrations in leachate were higher in the presence of ferns. Fern arsenic uptake was always greater than loss by leaching. Most arsenic (>66%) accumulated in P. vittata appeared of rhizosphere origin. In the medium-textured soil with more clay and higher nutrient content, successful iron scavenging increased arsenic release from soil for leaching, but transpiration curtailed leaching.

59 BASIC BIOLOGICAL SCIENCES↗

The effects of warming and soil chemistry on bacterial community structure in Arctic tundra soils

Soil microbial communities in the Arctic play a critical role in regulating the global carbon (C) cycle. Vast amounts of C are stored in northern latitude soils, and rising temperatures in the Arctic threaten to thaw permafrost, making relatively inaccessible C sources more available for mineralization by soil microbes. Few studies have characterized how microbial community structure responds to thawing permafrost in the context of varying soil chemistries associated with contrasting tundra landscapes. Here, we subjected active layer and permafrost soils from upland and lowland tundra sites on the North Slope of Alaska to a soil-warming incubation experiment and compared soil bacterial community profiles (obtained by 16S rRNA amplicon sequencing) before and after incubation. The influence of soil composition (characterized by mid-infrared [MIR] spectroscopy) on bacterial community structure and class abundance was analyzed using redundancy and correlation analyses. We found increased abundances of Alphaproteobacteria, Gammaproteobacteria, and Bacteroidetes [Sphingobacteriia] post incubation, particularly in permafrost soils. The categorical descriptors site and soil layer had the most explanatory power in our predictive models of bacterial community structure, highlighting the close relationship between soil bacteria and the soil environment. Specific soil chemical attributes characterizing the soil environments that were found to be the best predictors include MIR spectral bands associated with inorganic C, silicates, amide II (C=N stretch), and carboxylics (C-O stretch), and MIR peak ratios representing C substrate quality. Overall, these results further characterize soil bacterial community shifts that may occur as a frozen environment with limited access to C sources, such as is found in undisturbed permafrost, transitions to warmer and more C-available environments, such as is predicted in thawing permafrost due to climate change.

16S rRNA sequencing↗

Pyrolysis temperature and soil depth interactions determine PyC turnover and induced soil organic carbon priming

Pyrogenic organic carbon (PyC) is a complex, heterogeneous class of thermally altered organic substrates, but its dynamics and how its behavior changes with soil depth remain poorly understood. We conducted a laboratory incubation study to investigate the interactive effects of pyrolysis temperature and soil depth on the turnover of PyC compared to its precursor wood and native SOC (NSOC). We incubated dual-labeled (13C and 15N) jack pine pyrogenic organic matter produced at 300 °C (PyC300), 450 °C (PyC450), and their precursor pine wood in a fine-loamy, mixed-conifer forest soil for 745 days. A mixture of surface (0–10 cm) and subsurface (50–70 cm) forest soils, with and without labeled biomass were incubated in the dark at 55% soil water field capacity and 25 °C. Total 13C from PyC and wood mineralized as 13C-CO2 (as % of C added to soil) declined with an increase in pyrolysis temperature as follows: 54 ± 7.7% for wood, 3.1 ± 0.2% for PyC300, and 0.94 ± 0.08% for PyC450. Furthermore, after 2 years, soil depth interacted with pyrolysis temperature to affect C turnover, with total wood C losses significantly declining from 70.6% in surface soils to 37.5% in subsurface soil, while total losses of PyC300 and PyC450 were unaffected by differences between surface and subsurface soils. Wood induced negative priming (i.e., decreased mineralization rates) in surface soil at days 3 and 60, while PyC300 induced positive priming (i.e., increased mineralization rates) in subsurface soil at day 60. After 2 years, unlabeled NSOC losses increased from 9.2 ± 0.8% of NSOC in unamended treatments to 16.5 ± = 2.6% of NSOC with PyC450 additions. Our results suggest that PyC pyrolyzed at a given temperature can mineralize at similar rates between soil depths, and high amounts of PyC450 in subsurface soils can stimulate NSOC losses. These findings indicate that soil depth imposes critical controls on PyC dynamics belowground.

13C-labeling↗

Quantifying pH buffering capacity in acidic, organic-rich Arctic soils: Measurable proxies and implications for soil carbon degradation

Dynamic pH change promoted by biogeochemical reactions in Arctic tundra soils can be a major control on the production and release of CO 2 and CH 4 , which contribute to rising global temperatures. Large quantities of soil organic matter (SOM) in these soils are susceptible to microbial decomposition, leading to pH changes during permafrost thaw. Soil pH buffering capacity (β) modulates the extent of pH change but has not been thoroughly studied and represented in predictive ecosystem scale biogeochemical models in Arctic tundra soils. In this study, we generated titration curves for 21 acidic tundra soils from three Arctic sites across northern Alaska, United States of America. Geochemical and hydrological soil properties were evaluated, and correlations with β were developed. Strong correlations between β and both gravimetric water content (Θ g ) (R 2 = 0.847, p < 0.001) and soil water retention (SWR) (R 2 = 0.849, p = 0.001) indicate that the ability of soil to retain water could be associated with its buffering properties. Correlations between β and soil organic carbon (SOC) and cation exchange capacity (CEC) were also explored, and relationships to SWR are discussed. These correlations were then used with existing soil databases reporting SOC, CEC, and SWR to estimate β across Alaska soils. We further demonstrated the quantitative relationships between β and the simulated rates of biogeochemical reactions and show that lower β leads to higher soil pH and more CH 4 production. Overall, our study provides simple proxies for β in Arctic soils and highlights the importance and implications of representing soil buffering in predictive models, thereby enabling quantitative coupling between pH dynamics associated with biogeochemical reactions. Integrating β into predictive models of Arctic biogeochemical cycling may reduce model uncertainty and further our understanding of permafrost SOM degradation accelerated by warming.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Study of Oak Ridge soils using BONCAT-FACS-Seq reveals that a large fraction of the soil microbiome is active

The ability to link soil microbial diversity to soil processes requires technologies that differentiate active subpopulations of microbes from so-called relic DNA and dormant cells. Measures of microbial activity based on various techniques including DNA labelling have suggested that most cells in soils are inactive, a fact that has been difficult to reconcile with observed high levels of bulk soil activities. We hypothesized that measures of in situ DNA synthesis may be missing the soil microbes that are metabolically active but not replicating, and we therefore applied BONCAT (Bioorthogonal Non Canonical Amino Acid Tagging) i.e. a proxy for activity that does not rely on cell division, to measure translationally active cells in soils. We compared the active population of two soil depths from Oak Ridge (TN) incubated under the same conditions for up to seven days. Depending on the soil, a maximum of 25 – 70% of the cells were active, accounting for 3-4 million cells per gram of soil type, which is an order of magnitude higher than previous estimates. The BONCAT positive cell fraction was recovered by fluorescence activated cell sorting (FACS) and identified by 16S rDNA amplicon sequencing. The diversity of the active fraction was a selected subset of the bulk soil community. Excitingly, some of the same members of the community were recruited at both depths independently from their abundance rank. On average, 86% of sequence reads recovered from the active community shared >97% sequence similarity with cultured isolates from the field site. Our observations are in line with a recent report that, of the few taxa that are both abundant and ubiquitous in soil, 45% are also cultured – and indeed some of these ubiquitous microorganisms were found to be translationally active. The use of BONCAT on soil microbiomes provides evidence that a large portion of the soil microbes can be active simultaneously. We conclude that BONCAT coupled to FACS and sequencing is effective for interrogating the active fraction of soil microbiomes in situ and provides new perspectives to link metabolic capacity to overall soil ecological traits and processes.

58 GEOSCIENCES↗