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DOE OSTI2024

Improved Characterization of Soil Organic Matter by Integrating FT-ICR MS, Liquid Chromatography Tandem Mass Spectrometry, and Molecular Networking: A Case Study of Root Litter Decay under Drought Conditions

Understanding of how soil organic matter (SOM) chemistry is altered in a changing climate has advanced considerably; however, most SOM components remain unidentified, impeding the ability to characterize a major fraction of organic matter and predict what types of molecules, and from which sources, will persist in soil. Here we present a novel approach to better characterize SOM extracts by integrating information from three types of analyses, and we deploy this method to characterize decaying root-detritus soil microcosms subjected to either drought or normal conditions. To observe broad differences in composition, we employed direct infusion Fourier-transform ion cyclotron resonance mass spectrometry (DI-FT-ICR MS). We complemented this with liquid chromatography tandem mass spectrometry (LC-MS/MS) to identify components by library matching. Since libraries contain only a small fraction of SOM components, we also used fragment spectral cosine similarity scores to relate unknowns and library matches through molecular networks. This integrated approach allowed us to corroborate DI-FT-ICR MS molecular formulas using library matches, which included fungal metabolites and related polyphenolic compounds. We also inferred structures of unknowns from molecular networks and improved LC-MS/MS annotation rates from ~5 to 35% by considering DI-FT-ICR MS molecular formula assignments. Under drought conditions, we found greater relative amounts of lignin-like vs condensed aromatic polyphenol formulas and lower average nominal oxidation state of carbon, suggesting reduced decomposition of SOM and/or microbes under stress. Our integrated approach provides a framework for enhanced annotation of SOM components that is more comprehensive than performing individual data analyses in parallel.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2024

Bridging 20 Years of Soil Organic Matter Frameworks: Empirical Support, Model Representation, and Next Steps

Abstract In the past few decades, there has been an evolution in our understanding of soil organic matter (SOM) dynamics from one of inherent biochemical recalcitrance to one deriving from plant‐microbe‐mineral interactions. This shift in understanding has been driven, in part, by influential conceptual frameworks which put forth hypotheses about SOM dynamics. Here, we summarize several focal conceptual frameworks and derive from them six controls related to SOM formation, (de)stabilization, and loss. These include: (a) physical inaccessibility; (b) organo‐mineral and ‐metal stabilization; (c) biodegradability of plant inputs; (d) abiotic environmental factors; (e) biochemical reactivity and diversity; and (f) microbial physiology and morphology. We then review the empirical evidence for these controls, their model representation, and outstanding knowledge gaps. We find relatively strong empirical support and model representation of abiotic environmental factors but disparities between data and models for biochemical reactivity and diversity, organo‐mineral and ‐metal stabilization, and biodegradability of plant inputs, particularly with respect to SOM destabilization for the latter two controls. More empirical research on physical inaccessibility and microbial physiology and morphology is needed to deepen our understanding of these critical SOM controls and improve their model representation. The SOM controls are highly interactive and also present some inconsistencies which may be reconciled by considering methodological limitations or temporal and spatial variation. Future conceptual frameworks must simultaneously refine our understanding of these six SOM controls at various spatial and temporal scales and within a hierarchical structure, while incorporating emerging insights. This will advance our ability to accurately predict SOM dynamics.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2021

Direct evidence for the role of microbial community composition in the formation of soil organic matter composition and persistence

The largest terrestrial carbon sink on earth is soil carbon stocks. As the climate changes, the rate at which the Earth’s climate warms depends in part on the persistence of soil organic carbon. Microbial turnover forms the backbone of soil organic matter (SOM) formation and it has been recently proposed that SOM molecular complexity is a key driver of stability. Despite this, the links between microbial diversity, chemical complexity and biogeochemical nature of SOM remain missing. Here we tested the hypotheses that distinct microbial communities shape the composition of SOM, and microbial-derived SOM has distinct decomposition potential depending on its community of origin. We inoculated microbial communities of varying diversities into a model soil matrix amended with simple carbon (cellobiose) and measured the thermal stability of the resultant SOM. Using a Rock-Eval ® ramped thermal analysis, we found that microbial community composition drives the chemical fingerprint of soil carbon. While diversity was not a driver of SOM composition, bacteria-only communities lead to more thermally labile soil C pools than communities with bacteria and fungi. Our results provide direct evidence for a link between microbial community structure, SOM composition, and thermal stability. This evidence demonstrates the relevance of soil microorganisms in building persistent SOM stocks.

Domeignoz-Horta, Luiz A. (ORCID:0000000346186253)↗
DOE OSTI2021

Arbuscular mycorrhizal trees cause a higher carbon to nitrogen ratio of soil organic matter decomposition via rhizosphere priming than ectomycorrhizal trees

Tree roots and their associated microbes can significantly influence soil organic matter (SOM) decomposition, i.e., the rhizosphere priming effect. This effect is expected to be greater in trees associated with arbuscular mycorrhizal (AM) fungi, which produce higher extracellular enzymes especially surrounding hyphae, than in trees associated with ectomycorrhizal (ECM) fungi. In this work, we selected five tree species associated with AM (Juglans mandshurica Maxim. and Cunninghamia lanceolata (Lamb.) Hook.) or ECM (Picea koraiensis Nakai, Quercus mongolica Fischer ex Turcz. and Larix kaempferi (Lamb.) Carriere). We grew tree seedlings inside of cores lined with different mesh sizes to investigate how roots, hyphae and exudates influence soil carbon (C) and nitrogen (N) mineralization via the rhizosphere priming effect, using a 13 C natural abundance approach and a 15 N pool dilution method, concurrently. We found that tree seedlings significantly accelerated soil C decomposition by on average 78%, i.e., positive priming, compared to unplanted control pots. AM-associated trees induced 2.1 times greater soil C decomposition than ECM-associated trees across all mesh sizes. In contrast, gross N mineralization did not differ between tree-mycorrhizal associations. Compared to ECM counterparts, AM-associated trees had higher C- and lower N-degrading enzyme activities. Consequently, AM-associated trees induced a significantly higher C:N ratio of SOM decomposition than their ECM counterparts, which could be associated with the differences in soil enzyme activities for C and N degradation. Further, for both AM- and ECM-associated trees, we found no significant influences of mesh size on soil C decomposition, suggesting that the rhizosphere priming effect of mycorrhizal symbiosis was predominantly driven by root exudates. We conclude that SOM decomposition caused by AM-associated trees may have a higher C:N ratio than that by ECM-associated trees mainly due to differences in microbial enzyme investment. Our findings imply that tree-mycorrhizal associations are capable of modulating soil biogeochemical cycling via the rhizosphere priming effect.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2020

Controls on Soil Organic Matter Degradation and Subsequent Greenhouse Gas Emissions Across a Permafrost Thaw Gradient in Northern Sweden

Warming-induced permafrost thaw could enhance microbial decomposition of previously stored soil organic matter (SOM) to carbon dioxide (CO 2 ) and methane (CH 4 ), one of the most significant potential feedbacks from terrestrial ecosystems to the atmosphere in a changing climate. The environmental parameters regulating microbe-organic matter interactions and greenhouse gas (GHG) emissions in northern permafrost peatlands are however still largely unknown. The objective of this work is to understand controls on SOM degradation and its impact on porewater GHG concentrations across the Stordalen Mire, a thawing peat plateau in Northern Sweden. Here, we applied high-resolution mass spectrometry to characterize SOM molecular composition in peat soil samples from the active layers of a Sphagnum-dominated bog and rich fen sites in the Mire. Microbe-organic matter interactions and porewater GHG concentrations across the thaw gradient were controlled by aboveground vegetation and soil pH. An increasingly high abundance of reduced organic compounds experiencing greater humification rates due to enhanced microbial activity were observed with increasing thaw, in parallel with higher CH 4 and CO 2 porewater concentrations. Bog SOM however contained more Sphagnum-derived phenolics, simple carbohydrates, and organic- acids. The low degradation of bog SOM by microbial communities, the enhanced SOM transformation by potentially abiotic mechanisms, and the accumulation of simple carbohydrates in the bog sites could be attributed in part to the low pH conditions of the system associated with Sphagnum mosses. We show that Gibbs free energy of C half reactions based on C oxidation state for OM can be used as a quantifiable measure for OM decomposability and quality to enhance current biogeochemical models to predict C decomposition rates. We found a direct association between OM chemical diversity and δ 13 C-CH 4 in peat porewater; where higher substrate diversity was positively correlated with enriched δ 13 C-CH 4 in fen sites. Oxidized sulfur-containing compounds, produced by Sphagnum, were further hypothesized to control GHG emissions by acting as electron acceptors for a sulfate-reducing electron transport chain, inhibiting methanogenesis in peat bogs. These results suggest that warming-induced permafrost thaw might increase organic matter lability, in subset of sites that become wetlands, and shift biogeochemical processes toward faster decomposition with an increasing proportion of carbon released as CH 4 .

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2021

Fire effects on the persistence of soil organic matter and long-term carbon storage

One paradigm in biogeochemistry is that frequent disturbance tends to deplete carbon (C) in soil organic matter (SOM) by reducing biomass inputs and promoting losses. However, disturbance by fire has challenged this paradigm because soil C responses to frequent and/or intense fires are highly variable, despite observed declines in biomass inputs. Here, we review recent advances to illustrate that fire-driven changes in decomposition, mediated by altered SOM stability, are an important compensatory process offsetting declines in aboveground biomass pools. Fire alters the stability of SOM by affecting both the physicochemical properties of the SOM and the environmental drivers of decomposition, potentially offsetting C lost via combustion, but the mechanisms affecting the SOM stability differ across ecosystems. Thus, shifting our focus from a top-down view of fire impacting C cycling via changes in plant biomass to a bottom-up view of changes in decomposition may help to elucidate counterintuitive trends in the response of SOM to burning. Finally, given that 70% of global topsoil C is in fire-prone regions, using fire to promote SOM stability may be an important nature-based climate solution to increase C storage.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Differences in soil organic matter between EcM- and AM-dominated forests depend on tree and fungal identity

As global change shifts the species composition of forests, we need to understand which species characteristics affect soil organic matter (SOM) cycling to predict future soil carbon (C) storage. Recently, whether a tree species forms a symbiosis with arbuscular (AM) versus ectomycorrhizal (EcM) fungi has been suggested as a strong predictor of soil C storage, but there is wide variability within EcM systems. In this study, we investigated how mycorrhizal associations and the species composition of canopy trees and mycorrhizal fungi related to the proportion of soil C and nitrogen (N) in mineral associations and soil C:N across four sites representing distinct climates and tree communities in the eastern US broadleaf forest biome. In two of our sites, we found the expected relationship of declining mineral-associated C and N and increasing soil C:N ratios as the basal area of EcM-associating trees increased. However, across all sites these soil properties strongly correlated with canopy tree and fungal species composition. Sites where the expected pattern with EcM basal area was observed were (1) dominated by trees with lower quality litter in the Pinaceae and Fagaceae families and (2) dominated by EcM fungi with medium-distance exploration type hyphae, melanized tissues, and the potential to produce peroxidases. Here, this observational study demonstrates that differences in SOM between AM and EcM systems are dependent on the taxa of trees and EcM fungi involved. Important information is lost when the rich mycorrhizal symbiosis is reduced to two categories.

mineral-associated organic matter↗
DOE OSTI2020

Influences of Hillslope Biogeochemistry on Anaerobic Soil Organic Matter Decomposition in a Tundra Watershed

In this work, we investigated rates and controls on greenhouse gas (CO 2 and CH 4 ) production in two contrasting water-saturated tundra soils within a permafrost-affected watershed near Nome, Alaska, United States. Three years of field sample analysis have shown that soil from a fen-like area in the toeslope of the watershed had higher pH and higher porewater ion concentrations than soil collected from a bog-like peat plateau at the top of the hillslope. The influence of these contrasting geochemical and topographic environments on CO 2 and CH 4 production was tested in soil microcosms by incubating both the organic- and mineral-layer soils anaerobically for 55 days. Additionally, nitrogen (as NH 4 Cl) was added to half of the microcosms to test potential effects of N limitation on microbial greenhouse gas production. We found that the organic toeslope soils produced more CO 2 and CH 4 , fueled by higher pH and higher concentrations of water-extractable organic C (WEOC). Our results also indicate N limitation on CO 2 production in the peat plateau soils but not the toeslope soils. Together these results suggest that the weathering and leaching of ions and nutrients from tundra hillslopes can increase the rate of anaerobic soil organic matter decomposition in downslope soils by (1) increasing the pH of soil porewater; (2) providing bioavailable WEOC and fermentation products such as acetate; and (3) relieving microbial N limitation through nutrient runoff. We conclude that the soil geochemistry as mediated by landscape position is an important factor influencing the rate and magnitude of greenhouse gas production in tundra soils.

58 GEOSCIENCES↗
DOE OSTI2023

Porewater constituents inhibit microbially mediated greenhouse gas production (GHG) and regulate the response of soil organic matter decomposition to warming in anoxic peat from a Sphagnum -dominated bog

Abstract Northern peatlands store approximately one-third of terrestrial soil carbon. Climate warming is expected to stimulate the microbially mediated degradation of peat soil organic matter (SOM), leading to increasing greenhouse gas (GHG; carbon dioxide, CO2; methane, CH4) production and emission. Porewater dissolved organic matter (DOM) plays a key role in SOM decomposition; however, the mechanisms controlling SOM decomposition and its response to warming remain unclear. The temperature dependence of GHG production and microbial community dynamics were investigated in anoxic peat from a Sphagnum-dominated peatland. In this study, peat decomposition, which was quantified by GHG production and carbon substrate utilization is limited by terminal electron acceptors (TEA) and DOM, and these controls of microbially mediated SOM degradation are temperature-dependent. Elevated temperature led to a slight decrease in microbial diversity, and stimulated the growth of specific methanotrophic and syntrophic taxa. These results confirm that DOM is a major driver of decomposition in peatland soils contains inhibitory compounds, but the inhibitory effect is alleviated by warming.

Song, Tianze (ORCID:0000000282060247)↗
DOE OSTI2021

Ecological stoichiometry as a foundation for omics-enabled biogeochemical models of soil organic matter decomposition

Coupled biogeochemical cycles drive ecosystem ecology by influencing individual-to-community scale behaviors; yet the development of process-based models that accurately capture these dynamics remains elusive. Soil organic matter (SOM) decomposition in particular is influenced by resource stoichiometry that dictates microbial nutrient acquisition (‘ecological stoichiometry’). Despite its basis in biogeochemical modeling, ecological stoichiometry is only implicitly considered in high-resolution microbial investigations and the metabolic models they inform. State-of-science SOM decomposition models in both fields have advanced largely separately, but they agree on a need to move beyond seminal pool-based models. This presents an opportunity and a challenge to maximize the strengths of various models across different scales and environmental contexts. To address this challenge, we contend that ecological stoichiometry provides a framework for merging biogeochemical and microbiological models, as both explicitly consider substrate chemistries that are the basis of ecological stoichiometry as applied to SOM decomposition. We highlight two gaps that limit our understanding of SOM decomposition: (1) understanding how individual microorganisms alter metabolic strategies in response to substrate stoichiometry and (2) translating this knowledge to the scale of biogeochemical models. We suggest iterative information exchange to refine the objectives of high-resolution investigations and to specify limited dynamics for representation in large-scale models, resulting in a new class of omics-enabled biogeochemical models. Assimilating theoretical and modelling frameworks from different scientific domains is the next frontier in SOM decomposition modelling; advancing technologies in the context of stoichiometric theory provides a consistent framework for interpreting molecular data, and further distilling this information into tractable SOM decomposition models.

Carbon cycling↗
DOE OSTI2025

Quantifying soil organic matter stock distribution and origin following over a century of maize-based cropping in the former tallgrass prairie region of central USA

Tallgrass prairie conversion to maize-based agriculture in central North America has resulted in substantial loss of soil organic carbon (SOC) in less than two centuries. However, evaluations of how management practices may mitigate SOC losses are generally limited in soil depth and/or duration, missing long-term SOC stock outcomes that manifest over timescales of decades or longer. To address this, we sampled soils in year 145 of the Morrow Plots experiment to (i) evaluate effects of crop rotation and fertility management on SOC stocks and (ii) distinguish prairie- versus maize-derived SOC after continuous maize cropping since 1876 using stable carbon isotope ( 13 C) natural abundance. Soil organic carbon stock by equivalent soil mass (ESM) was + 30.7 Mg C ha −1 (+31.7 %) higher under maize-oat-alfalfa than continuous maize, but similar between maize-soybean and continuous maize. NPK fertilization and manuring did not influence SOC stocks by ESM. Response of SOC stocks at 15 cm depth intervals to NPK fertilization varied by depth and crop rotation, with lower SOC stocks at 30–45 cm under continuous maize and maize-soybean. Maize-derived C ranged 19.5–59.6 % of SOC stock across depths, indicating the majority of SOC was still derived from tallgrass prairie even after 145 years of continuous maize cropping. Our results confirm the potential of diversified crop rotation for minimizing SOC losses relative to tallgrass prairie at the supracentennial scale, and highlight the importance of relic prairie soil organic matter for future crop production in central North America.

crop rotation↗
DOE OSTI2026

Shrub and sedge rhizosphere communities display distinct affinities toward exudates and soil organic matter degradation: a quantitative stable isotope probing analysis

Warming temperatures are accelerating permafrost thaw and changing tundra vegetation, where woody shrubs are displacing sedges. Shrubs, such as Betula nana, and sedges, such as Eriophorum vaginatum, exhibit distinct life strategies including unique root-associated, or rhizosphere microbial communities. As permafrost thaws it unlocks previously unavailable carbon and nutrient sources resulting in deeper roots and a translocation of rhizosphere communities. Because permafrost microbial communities contain lower diversity and biomass than rhizosphere communities, the coalescence of rhizosphere and permafrost microbial communities could alter soil organic matter (SOM) degradation rates and increase greenhouse gas emissions. To identify metabolic strategies across distinct rhizosphere and permafrost microbial communities we conducted an isotope tracing incubation experiment. We inoculated thawed permafrost with shrub and sedge rhizosphere communities while adding exudates or water daily and compared this to an uninoculated control. After 46 days, we spiked samples with 18O enriched water or 13C enriched exudates and measured isotope incorporation into microbial DNA with quantitative stable isotope probing (qSIP). Our results indicate that exudate additions had little effect on uninoculated permafrost communities but the addition of exudates and rhizosphere inoculants had a compounding effect on respiration rates. We found that soils inoculated with shrub rhizosphere communities contained a mixture of exudate and SOM degraders while soils inoculated with sedge rhizosphere communities contained mainly SOM degraders. Finally, we found that individual microbial taxa exhibited maximum growth rates under specific combinations of microbial inoculant communities and exudate addition treatments. Our results reveal that microbial niches are strongly influenced by substrate preferences and community context, and suggest that a reduction in sedges and an expansion of shrubs may provide a mechanism by which permafrost carbon losses are mitigated through corresponding shifts in microbial communities and their substrate preferences.

Schaefer, Sean R [Natural Resources and Earth Syst↗
DOE OSTI2021

Soil organic matter is principally root derived in an Ultisol under oak forest

To a large degree, the sources and stability of soil organic carbon remain poorly constrained. A clear understanding of links among the components of the soil C cycle is hampered by the complexity of the system as well as challenges associated with partitioning bulk soil C into meaningful fractions. A large accidental 14 CO 2 release at the Oak Ridge Reservation in Tennessee, USA provided a strong label pulse into adjacent, well-studied oak forests, resulting in highly elevated Δ 14 C values in leaf litter (~1000‰) and roots (~260–450‰). A four-year manipulative study was conducted to determine the relative contribution of litter versus roots to the bulk mineral soil C pool, as well as to free light, occluded light and heavy fractions. The heavy fraction was further split into fractions with densities of 1.7–2.4 g cm –3 and >2.4 g cm –3 to test the homogeneity of the mineral-associated fraction of C. Substantial concentrations of label were detected in all soil fractions within a year of the 14 CO 2 release, indicating rapid incorporation of newly fixed photosynthates in all fractions of soil organic C, regardless of differences in stability inferred by previous work. This rapid incorporation of label occurred only in treatments where roots were labeled, indicating that roots are the major source of inputs to mineral soil C stocks at these sites. Separation of the heavy fraction into subfractions of intermediate (1.7–2.4 g cm –3 ) and high (>2.4 g cm –3 ) density indicated that both subfractions incorporated label at similar rates, despite significant differences in degree of microbial processing. In general, the rate of label incorporation suggested a much faster turnover for all fractions than indicated by natural radiocarbon abundance values. This study suggests that within each soil fraction there are portions of slow-cycling and fast-cycling materials, and the determination of an average turnover time or mean age is dependent on experimental approach. The rapid incorporation of label into all fractions within a year regardless of inferred stability implies a high degree of heterogeneity in all fractions regardless of how finely the soils are partitioned. Further refinement of the nature and drivers of this heterogeneity could yield important insights into the soil C cycle.

58 GEOSCIENCES↗
DOE OSTI2023

Calcium promotes persistent soil organic matter by altering microbial transformation of plant litter

Abstract Calcium (Ca) can contribute to soil organic carbon (SOC) persistence by mediating physico-chemical interactions between organic compounds and minerals. Yet, Ca is also crucial for microbial adhesion, potentially affecting colonization of plant and mineral surfaces. The importance of Ca as a mediator of microbe-mineral-organic matter interactions and resulting SOC transformation has been largely overlooked. We incubated 44 Ca labeled soils with 13 C 15 N labeled leaf litter to study how Ca affects microbial transformation of litter and formation of mineral associated organic matter. Here we show that Ca additions promote hyphae-forming bacteria, which often specialize in colonizing surfaces, and increase incorporation of litter into microbial biomass and carbon use efficiency by approximately 45% each. Ca additions reduce cumulative CO 2 production by 4%, while promoting associations between minerals and microbial byproducts of plant litter. These findings expand the role of Ca in SOC persistence from solely a driver of physico-chemical reactions to a mediator of coupled abiotic-biotic cycling of SOC.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Microscale spatial distribution and soil organic matter persistence in top and subsoil

The spatial distribution of organic substrates and microscale soil heterogeneity significantly influence organic matter (OM) persistence as constraints on OM accessibility to microorganisms. However, it is unclear how changes in OM spatial heterogeneity driven by factors such as soil depth affect the relative importance of substrate spatial distribution on OM persistence. This work evaluated the decomposition and persistence of 13 C and 15 N labeled water-extractable OM inputs over 50 days as either hotspot (i.e., pelleted in 1–2 mm-size pieces) or distributed (i.e., added as OM < 0.07 μm suspended in water) forms in topsoil (0–0.2 m) and subsoil (0.8–0.9 m) samples of an Andisol. We observed greater persistence of added C in the subsoil with distributed OM inputs relative to hotspot OM, indicated by a 17% reduction in cumulative mineralization of the added C and a 10% higher conversion to mineral-associated OM. A lower substrate availability potentially reduced mineralization due to OM dispersion throughout the soil. NanoSIMS (nanoscale secondary ion mass spectrometry) analysis identified organo-mineral associations on cross-sectioned aggregate interiors in the subsoil. On the other hand, in the topsoil, we did not observe significant differences in the persistence of OM, suggesting that the large amounts of particulate OM already present in the soil outweighed the influence of added OM spatial distribution. Here, we demonstrated under laboratory conditions that the spatial distribution of fresh OM input alone significantly affected the decomposition and persistence of OM inputs in the subsoil. On the other hand, spatial distribution seems to play a lower role in topsoils rich in particulate OM. The divergence in the influence of OM spatial distribution between the top and subsoil is likely driven by differences in soil mineralogy and OM composition.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Soil organic matter diagenetic state informs boreal forest ecosystem feedbacks to climate change

The fate of soil organic carbon (SOC) in boreal forests is dependent on the integrative ecosystem response to climate change. For example, boreal forest productivity is often nitrogen (N) limited, and climate warming can enhance N cycling and primary productivity. However, the net effect of this feedback on the SOC reservoir and its longevity with climate change remain unclear due to difficulty in detecting small differences between large and variable carbon (C) fluxes needed to determine net changes in soil reservoirs. The diagenetic state of SOC – resulting from the physicochemical and biological transformations that alter the original biomolecular composition of detrital inputs to soil over time – is useful for tracing the net response of SOC at the timescales relevant to climate change not usually discernible from fluxes and stocks alone. Here, we test two hypotheses using a mesic boreal forest climate transect: (1) the SOC diagenetic state is maintained across this climosequence, and (2) the maintenance of the SOC diagenetic state is a consequence of coupled soil C and N cycling, signifying the role of enhanced N cycling supporting SOC inputs that maintain SOC stocks within the warmer-climate forests. Shifts in nonvascular to vascular plant inputs with climate observed in these and other boreal forests highlighted the need to carefully separate biogeochemical indicators of SOC source from those signifying diagenetic alteration. We thus evaluated and applied lignin biomarkers to assess the diagenetic alteration of SOC in these boreal forest organic soils and directly compared the lignin diagenetic state with that of soil organic nitrogen (SON) assessed through amino acid composition. The lignin diagenetic state remained constant across the climate transect, indicating a balance between the input and removal of lignin in these mesic boreal forests. When combined with previous knowledge of these forest ecosystems, including the diagenetic state of SON and direct measures of C fluxes and stocks, the results indicate a coupled increase in C and N cycling with climate warming that supports forest productivity and maintains SOC stocks. This balance could markedly shift as other factors begin to limit forest productivity (e.g., trace nutrients, water) with further climate change or affect forest nutrient allocation (e.g., forest age or compositional change). Further application of the approach presented here could be used to detect the limits of this and other ecosystem–climate feedbacks, by providing a tractable and parameterizable index of the lignin state across large spatial scales, necessary for ecosystem-scale parameterizations.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2023

Effects of root litter traits on soil organic matter dynamics depend on decay stage and root branching order

Decaying roots contribute disproportionately to soil organic carbon (SOC) stocks in many ecosystems, yet few studies have examined the afterlife effects of root trait variability on the formation, stability and mineralization of SOC. In order to assess how root functional class (absorptive vs. transport) and mycorrhizal association (ectomycorrhizal vs. arbuscular mycorrhizal fungi) affect both formation and loss of SOC, we decayed lower-order absorptive (orders 1–2) and higher-order transport (orders 3–4) roots from six arbuscular mycorrhizal (AM) and six ectomycorrhizal (EcM) associated hardwood tree species in an isotopically distinct soil under laboratory conditions. We found that root litter effects on SOC differed significantly among root functional classes and mycorrhizal types. AM absorptive and transport roots (high in nitrogen and soluble compounds) contributed more to the formation of new mineral-associated organic carbon (MAOC), but also contributed to higher mineralization of native SOC (via priming effects). Moreover, absorptive AM roots formed MAOC gradually and rates saturated, whereas ECM roots formed MAOC more quickly, despite slower decay (via greater formation efficiency). Altogether, these findings demonstrate how different combinations of root traits might contribute to soil C balance, and that failure to account for the chemical and morphological heterogeneity of roots, as well their interactions with microbes over time, may lead to incorrect projections of SOC stocks and turnover.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Soil organic matter, tree communities, and fungal communities across mycorrhizal gradients in the Eastern United States

We collected these data to investigate how mycorrhizal associations are related to soil C and N across four sites in the eastern U.S. broadleaf forest biome, which capture broad variability in climate and tree species. Our sites represented each of the four ecoregions in the eastern United States temperate forest—warm continental (New Hampshire), hot continental (Wisconsin), Prairie (Illinios), and subtropical (Georgia). This breadth naturally provided tree species diversity and allowed us to also investigate how soil C and N are related to canopy tree and EcM fungal community composition. Specifically, we analyzed the effects of mycorrhizal association, canopy tree family, and EcM fungal taxa on the proportion of C and N associated with soil minerals (MAOM) and bulk soil C:N. These are plot level data from forests in New Hampshire, Georgia, Wisconsin, and Illinois. The plots are 10 m in radius and vary across gradients of ectomycorrhizal tree basal area. Within each plot, data include the basal area of all trees >2 cm in diameter identified to species, the abundance of different fungal taxa based on ITS sequences, and the %C, %N, C:N of bulk soil and of soil density fractions (free light, occluded light, and particulate) as well as the proportion of soil C and N within each fraction. The soil data (both organic matter and fungal communities) are from the top 10 cm of the mineral soil. This also includes a meta-analysis of leaf litter k and C:N values for the tree species present in the forests.The "Tree_BA_ALL.csv" files include the basal area of all trees >2 cm in diameter at breast height within our 10 cm radii plots. The "Soil_Data_Final.csv" file contains the soil carbon and nitrogen in bulk and density fractions (concentrations as well as proportion for free light fraction (fLF), occluded light fraction (oLF), and heavy or dense fraction (HF), oxalate extractable iron and aluminum concentrations, as well as the geographic coordinates of each plot's location. The "FungiNonRelative.csv" file contains the abundances of fungal taxa identified via ITS sequencing for each plot (with the exception of 4 plots from GA where there was not enough quality DNA to extract).The "R_ReadyK.csv" and "R_Ready_CN.csv" files contain leaf litter decomposition constant (k) and C:N values from the literature for the tree species found at our sites.

54 ENVIRONMENTAL SCIENCES↗