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Increasing climatic pressures such as drought and flooding challenge agricultural systems and their management globally. How agricultural soils respond to soil water extremes will influence biogeochemical cycles of carbon and nitrogen in these systems. We investigated the response of soils from long-term agricultural field sites under varying crop rotational complexity to either drought or flooding stress. Focusing on these contrasting stressors separately, we investigated soil heterotrophic respiration during single and repeated stress cycles in soils from four different sites along a precipitation gradient (Colorado, MAP 421 mm; South Dakota, MAP 580 mm; Michigan, MAP 893 mm; Maryland, MAP 1192 mm); each site had two crop rotational complexity treatments. At the driest (Colorado) and wettest (Maryland) of these sites, we also analyzed microbial biomass, six potential enzyme activities, and N2O production during and after individual and repeated stress cycles. In general, we found site specific responses to soil water extremes, irrespective of crop rotational complexity and precipitation history. Drought usually caused more severe changes in respiration rates and potential enzyme activities than flooding. All soils returned to control levels for most measured parameters as soon as soils returned to control water levels following drought or flood stress, suggesting that the investigated soils were highly resilient to the applied stresses. The lack of sustained responses following the removal of the stressors may be because they are well in the range of natural in situ soil water fluctuations at the investigated sites. Without the inclusion of plants in our experiment, we found that irrespective of crop rotation complexity, soil and microbial properties in the investigated agricultural soils were more resistant to flooding but highly resilient to drought and flooding during single or repeated stress pulses.

In temperate soil systems, microbial biomass often increases during winter and decreases again in spring. This build-up and release of microbial carbon could potentially lead to a stabilization of soil carbon during winter times. Whether this increase is caused by changes in microbial physiology, in community composition or by changed substrate allocation within microbes or communities is unclear. In a laboratory incubation study, we looked into microbial respiration and growth, as well as microbial glucose uptake and carbon resource partitioning in response to cooling. Soils taken from a temperate forest and an agricultural system in October 2020, were cooled down from field temperature of 11 °C to 1 °C. We determined microbial growth using 18O-incorporation into DNA immediately after cooling and after an acclimation phase of 7 days; in addition, we traced 13C-labelled glucose into microbial biomass, CO2 respired from the soil, and into microbial phospholipid fatty acids (PLFAs). Our results show that the studied soil microbial communities responded immediately to soil cooling. Independent of soil type and acclimation period, total respiration, as well as 18O based growth, and thus cell division were strongly reduced when soils were cooled from 11 °C to 1 °C, while glucose uptake and glucose-derived respiration were unchanged. We found that microbes increased the investment of glucose-derived carbon in unsaturated phospholipid fatty acids at colder temperatures. Since unsaturated fatty acids retain fluidity at lower temperatures compared to saturated fatty acids, this could be interpreted as a precaution to reduced temperatures. Together with the maintained glucose uptake and reduced cell division, our findings show an immediate response of soil microorganisms to soil cooling, potentially to prepare for freeze-thaw events. The discrepancy between C uptake and cell division, further hints at a mechanism that could explain previously observed high microbial biomass carbon in temperate soils in winter.

Soil microbial growth, respiration and carbon use efficiency (CUE) are essential parameters to understand, describe and model the soil carbon cycle. While seasonal dynamics of microbial respiration are well studied, little is known about how microbial growth and CUE change over the course of a year, especially outside the plant growing season. In this study we measured soil microbial respiration, growth and biomass in an agricultural field and a deciduous forest 16 times over the course of two years. We sampled plots, at which harvest residues or leaf litter were either incorporated or removed. We observed strong seasonal variations of microbial respiration, growth and biomass. All microbial parameters were significantly higher at the forest site, which contained 3.5% organic C compared to the agricultural site with 0.9% organic C. CUE also varied strongly but was overall significantly higher at the agricultural site ranging from 0.1 to 0.7 compared to the forest site where CUE ranged from 0.1 to 0.6. We found that microbial respiration and to a lesser extent microbial growth followed the seasonal dynamics of soil temperature. Microbial growth was further affected by plant or foliage presence. At low temperatures in winter, both microbial respiration and growth rates were lowest. Due to higher temperature sensitivity of microbial respiration, CUE showed the highest values in the coldest months. Microbial biomass C was also strongly increased in winter. Surprisingly, this winter peak was not connected to high microbial growth or an increase in DNA content. This suggests that microorganisms accumulated osmo- or cryoprotectants but did not divide. This microbial winter bloom and following decline, where C is released and can be stabilized, could constitute the main season for C sequestration in temperate soil systems. Highly variable CUE, and the fact that CUE is calculated from independently controlled microbial respiration and growth, ask for great caution when CUE is used to describe soil microbial physiology, soil C dynamics or C sequestration. Instead, microbial respiration, microbial growth and biomass should rather be investigated individually to better understand the soil C cycle.

A longstanding assumption of glucose tracing experiments is that all glucose is microbially utilized during short incubations of ≤2 days to become microbial biomass or carbon dioxide. Carbon use efficiency (CUE) estimates have consequently ignored the formation of residues (non-living microbial products) although such materials could represent an important sink of glucose that is prone to stabilization as soil organic matter. We examined the dynamics of microbial residue formation from a short tracer experiment with frequent samplings over 72 h, and conducted a meta-analysis of previously published glucose tracing studies to assess the generality of these experimental results. Both our experiment and meta-analysis indicated 30–34% of amended glucose-C (13C or 14C) was in the form of residues within the first 6 h of substrate addition. We expand the conventional efficiency calculation to include residues in both the numerator and denominator of efficiency, thereby deriving a novel metric of the potential persistence of glucose-C in soil as living microbial biomass plus residues (‘carbon stabilization efficiency’). This new metric indicates nearly 40% of amended glucose-C persists in soil 180 days after amendment, the majority as non-biomass residues. Starting microbial biomass and clay content emerge as critical factors that positively promote such long term stabilization of labile C. Rapid residue production supports the conclusion that non-growth maintenance activity can illicit high demands for C in soil, perhaps equaling that directed towards growth, and that residues may have an underestimated role in the cycling and sequestration potential of C in soil.

Permafrost-affected soils in the northern circumpolar region store more than 1,000 Pg soil organic carbon (OC), and are strongly vulnerable to climatic warming. However, the extent to which changing soil environmental conditions with permafrost thaw affects different compounds of soil organic matter (OM) is poorly understood. Here, we assessed the fate of lignin and non-cellulosic carbohydrates in density fractionated soils (light fraction, LF vs. heavy fraction, HF) from three permafrost regions with decreasing continentality, expanding from east to west of northern Siberia (Cherskiy, Logata, Tazovskiy, respectively). In soils at the Tazovskiy site with thicker active layers, the LF showed smaller OC-normalized contents of lignin-derived phenols and plant-derived sugars and a decrease of these compounds with soil depth, while a constant or even increasing trend was observed in soils with shallower active layers (Cherskiy and Logata). Also in the HF, soils at the Tazovskiy site had smaller contents of OC-normalized lignin-derived phenols and plant-derived sugars along with more pronounced indicators of oxidative lignin decomposition and production of microbial-derived sugars. Active layer deepening, thus, likely favors the decomposition of lignin and plant-derived sugars, that is, lignocelluloses, by increasing water drainage and aeration. Our study suggests that climate-induced degradation of permafrost soils may promote carbon losses from lignin and associated polysaccharides by abolishing context-specific preservation mechanisms. However, relations of OC-based lignin-derived phenols and sugars in the HF with mineralogical properties suggest that future OM transformation and carbon losses will be modulated in addition by reactive soil minerals. publishedVersion

Soil carbon models typically scale decomposition linearly with soil carbon (C) concentration, but this linear relationship has not been experimentally verified. Here we investigated the underlying biogeochemical mechanisms controlling the relationships between soil C concentration and decomposition rates. We incubated a soil/sand mixture with increasing amounts of finely ground plant residue in the laboratory at constant temperature and moisture for 63 days. The plant residues were rye (Secale cereale, C/N ratio of 23) and wheat straw (Triticum spp., C/N ratio of 109) at seven soil C concentrations ranging from 0.38 to 2.99%. We measured soil respiration, dissolved organic carbon (DOC) concentrations, microbial biomass, and potential enzyme activities over the course of the incubation. Rye, which had higher N and DOC contents, lost 6 to 8 times more C as CO2 compared to wheat residue. Under rye and wheat amendment, absolute C losses as CO2 (calculated per g dry soil) increased linearly with C concentration while relative C losses as CO2 (expressed as percent of initial C) increased with C concentration following a quadratic function. In low C concentration treatments (0.38–0.79% OC), DOC decreased gradually from day 3 to day 63, microbial C increased towards the end in the rye treatment or decreased only slightly with straw amendment, and microbes invested in general enzymes such as proteases and oxidative enzymes. At increasing C levels, enzyme activity shifted to degrading cellulose after 15 days and degrading microbial necromass (e.g. chitin) after 63 days. At the highest C concentrations (2.99% OC), microbial biomass peaked early in the incubation and remained high in the rye treatment and decreased only slightly in the wheat treatment. While wheat lost C as CO2 constantly at all C concentrations, respiration dynamics in the rye treatment strongly depended on C concentration. Our results indicate that litter quality and C concentration regulate enzyme activities, DOC concentrations, and microbial respiration. The potential for non-linear relationships between soil C concentration and decomposition may need to be considered in soil C models and soil C sequestration management approaches.

Fertilization experiments provide insights into elemental imbalances in soil microbial communities and their consequences for soil nutrient cycling. By addition of selected nutrients, other nutrients become deficient and limiting for soil microorganisms as well as for plants. In this study we focused on microbial nitrogen (N) cycling in a long-term nutrient manipulation experiment. In many soils, the rate-limiting step in N cycling is depolymerization of high-molecular-weight nitrogen compounds (e.g., proteins) to oligomers (e.g., peptides) and monomers (e.g., amino acids) rather than the subsequent steps of mineralization (ammonification) and nitrification. The aim of our study was to determine whether nutrient deficiency directly or indirectly – via changes in plant carbon (C) inputs - affects soil microbial N processing.We collected soil samples from a fertilization experiment, established in 1946 on a hay meadow close to Admont (Styria, Austria). The field experiment consisted of a full factorial combination of inorganic N, P, and K fertilization and a control with no fertilizers. Furthermore, liming (Ca-addition) and organic fertilizer application treatments (solid manure and liquid slurry) were established. In the experiment, plant biomass is harvested three times per year, inducing strong nutrient limitation in plots that have not received nutrient additions (fully deficient or deficient in a single element). We determined gross rates of microbial protein depolymerization, N-mineralization and nitrification via isotope pool dilution assays with 15N-labeled amino acids, NH4+, and NO3-. We hypothesized that N deficiency (lack of N fertilization) would stimulate microbial N mining (depolymerization), and reduce subsequent N mineralization and nitrification. In contrast, we expected that organic fertilization would alleviate microbial C and N limitations, reducing N depolymerization rates and increasing mineralization and nitrification.Our results show that organically fertilized and limed soils have significantly lower gross protein depolymerization rates than plots receiving inorganic N. No significant differences were found comparing gross N-mineralization and gross nitrification rates across the different treatments. Given the higher rates of protein depolymerization in inorganically fertilized soils as compared to organically fertilized and limed soils, microbial N processes seem to be controlled by plant C input and/or soil pH rather than by direct soil nutrient availability. However, depolymerization of macromolecular N does not only supply N to the soil microbial community but also organic C. Thus, the reduced plant C input compared to fully fertilized soils may have caused microorganisms to increase their mining for a C-containing energy source, thereby increasing protein depolymerization rates. In summary, this study suggests that long term nutrient deficiency or nutrient imbalances may affect soil nutrient cycling indirectly by changing plant C inputs (via reduced primary production) and/or changing soil pH, rather than directly, by nutrient availability. This further indicates that soil microbial communities are rather C than nutrient limited.

The raise of atmospheric CO2 concentrations, with consequent increase in global warming and the likelihood of severe droughts, is altering the terrestrial biogeochemical carbon (C) cycle, with potential feedback to climate change. Microbial physiology, i.e. growth, turnover and carbon use efficiency, control soil carbon fluxes to the atmosphere. Thus, improving our ability to accurately quantify microbial physiology, and how it is affected by climate change, is essential. Recent advances in the field have allowed the quantification of community-level microbial growth and carbon use efficiency in dry conditions via an 18O water vapor equilibration technique, allowing for the first time to evaluate microbial growth rates under drought conditions.We modified the water vapor equilibration method using 2H-labelled water to estimate microbial community growth via deuterium incorporation into fatty acids. First, we verified that a rapid equilibration of 2H with soil water is possible. Then, we applied this approach to soil samples collected from a long-term climate change experiment (https://www.climgrass.at/) where warming, elevated atmospheric CO2 (eCO2) and drought are manipulated in a full factorial combination. Samples were taken in the field during peak drought and one week after rewetting. We used a high-throughput method to extract phospho- and neutral- lipid fatty acids (PLFA and NLFA) and we measured 2H enrichment in these compounds via GC-IRMS.Our results show that within 48 h, 2H in water vapor was in equilibrium with soil water and was detectable in microbial PLFA and NLFAs. We were able to quantify growth rates for different groups of microorganisms (Gram-positive, Gram-negative, Fungi and Actinobacteria) and calculate community level carbon use efficiency. We showed that a reduction of carbon use efficiency in the combined warming + eCO2 treatment was caused by a reduced growth of fungi and overall higher respiration rates. During drought, all groups showed a reduction in growth rates, albeit the reduction was stronger in bacteria than in fungi. Moreover, fungi accumulated high amounts of 2H into NLFAs, representing up to one third of the amount in PLFAs and indicating enhanced investment into storage compounds. This investment was still higher than in control plots two days after rewetting and returned to control levels within a week.Our study demonstrates that climate change can have strong effects on microbial physiology, with group-specific responses to different climate change factors. Our approach has the benefit of using fatty acid biomarkers to improve resolution into community level growth responses to climate change. This allowed a quantification of group-specific growth rates and concomitantly a measurement of investment into reserve compounds.

In temperate soil systems microbial biomass often increases during winter and decreases again in spring. This build up and release of microbial carbon could potentially lead to a build-up of stabilized soil carbon during winter times. The mechanism behind the increase in microbial carbon is not well understood. In this laboratory incubation study, we looked into microbial physiology as well as microbial glucose uptake and partitioning during cooling. Soils from a temperate forest and agricultural system were cooled down from field temperature of 11°C to 1°C. We added 13C-labelled glucose immediately and after an acclimation phase of 7 days and traced the 13C into microbial biomass, CO2 respired from the soil and phospholipid fatty acids. In addition we determined microbial growth using 18O-incorporation into DNA.First results show that while total respiration was strongly reduced when soils were cooled, glucose-derived respiration was as high in soils at 1°C as at 11°C. The same general pattern was found in soils during fast cooling and after an acclimation phase in agricultural and forest soils. We also saw an increased investment of glucose-derived carbon in unsaturated PLFAs. Since unsaturated fatty acids retain fluidity at lower temperatures compared to saturated fatty acids, this could be interpreted as precaution to reduced temperatures and potential freezing.Our results show a distinct response of the soil microbial community to cooling. The maintained glucose-derived respiration and incorporation into PLFAs at low temperatures compared to field temperature might indicate a preferential use of labile C forms during cooling. Moreover, the 13C incorporation into PLFAs may signal the buildup of cooling resistant cell membranes. These findings will be discussed with results from the 13C label tracing into microbial biomass, extractable organic carbon and total soil carbon as well as data on microbial growth and carbon use efficiency.

Eucalyptus plantations have replaced other (agro)ecosystems over 5.6 Mha in Brazil. While these plantations rapidly accumulate carbon (C) in their biomass, the C storage in living forest biomass is transient, and thus, longer‐term sustainability relies on sustaining soil organic matter (SOM) stocks. A significant amount of harvest residues (HR) is generated every rotation and can yield SOM if retained in the field. Yet, there is little information on how managing eucalyptus HR changes SOM dynamics. We used isotopic and molecular approaches in a 3‐yr field decomposition experiment where a native grassland has been replaced by eucalyptus plantations to assess how HR management practices influence content and chemistry of two distinct SOM fractions [particulate (POM) and mineral‐associated organic matter (MAOM)] at two soil depths (0–1 and 1–5 cm). The management practices investigated were HR removal (−R), only bark removal (−B), and retention of all HR (including bark, +B), combined with two levels of nitrogen (N) fertilization [0 (−N) and 200 (+N) kg/ha]. N fertilization inhibited HR decomposition (P = 0.0409), while bark retention had little effect (P = 0.1164). Retaining HR, especially with bark, increased POM‐C and MAOM‐C content (2.1‐ and 1.2‐fold, respectively), decreased POM‐δ13C (1.2‐fold), and increased inorganic N retention (1.7‐fold) compared with plots where HR had been removed. Inorganic N applications, however, diminished the positive impacts of bark retention. Although the influence of HR management was most pronounced in POM, retaining HR reduced potential soil C mineralization by up to 20%. POM and MAOM chemistry shifted over time and revealed distinct influence of HR on the formation of these fractions. We demonstrate that HR management alters SOM dynamics and that retaining HR, particularly including bark, enhances SOM retention. With continuing conversion of native grassland ecosystems to eucalyptus, long‐term sustainability will require careful HR and fertilizer management to balance total biomass harvest with sustaining belowground SOM concentrations.

As the global hydrological cycle intensifies with future warming, more severe droughts will alter the terrestrial biogeochemical carbon (C) cycle. As soil microbial physiology controls the large fluxes of C from soil to the atmosphere, improving our ability to accurately quantify microbial physiological parameters in soil is essential. However, currently available methods to determine microbial C metabolism in soil require the addition of water, which makes it practically impossible to measure microbial physiology in dry soil samples without stimulating microbial growth and respiration (namely, the “Birch effect”).We developed a new method based on in vivo 18O water vapor equilibration to minimize soil re-wetting effects. This method allows the isotopic labelling of soil water without any liquid water or dissolved substrate addition to the sample. This was compared to the main current method (18O-water application method) in soil samples either at near-optimal water holding capacity or in air dry soils. We generated time curves of the isotopic equilibration between liquid soil water and water vapor and calculated the average atom percent 18O excess over incubation time, which is necessary to calculate microbial growth rates. We tested isotopic equilibration patterns in nine different soils (natural and artificially constructed ones) covering a wide range of soil texture and organic matter content. We then measured microbial growth, respiration and carbon use efficiency in three natural soils (either dry or at near-optimal water holding capacity). The proposed 18O vapor equilibration method provides similar results as the currently widely used method of liquid 18O water addition to determine microbial growth when used a near-optimal water holding capacity. However, when applied to dry soils the liquid 18O water addition method overestimated growth by up to 250%, respiration by up to 500%, and underestimated carbon use efficiency by up to 40%.Finally, we applied the new method to undisturbed biocrust samples, at field water content (1-3%), and show for the first time real microbial growth rates and CUE values in such arid ecosystems. We describe new insights into biogeochemical cycling of C that the new method can help uncover and consider the wide range of questions regarding microbial physiology and its response to global change that can now be proposed and addressed.

Conservation or regenerative agriculture, i.e. reduction of mechanical soil disturbance, introduction of crop rotations, and especially cover crops as a form of natural soil amendment, has been shown to increase soil organic matter contents as well as soil health. One mechanism behind the increase in organic carbon under regenerative agriculture could be an increase in microbial biomass, as well as an enhanced carbon use efficiency (CUE) of the soil microorganisms in these systems. Such changes in microbial biomass and activity could also influence soil nitrogen (N) cycling. Here we show first results of on-farm research at four sites in Austria comparing crop fields under regenerative agriculture practices with conventional practices, and nearby perennial grasslands at each site. The four sites span different climate gradients, soil types and textures.Soil organic carbon (SOC) content ranged from 1 to 2.3% in the agricultural soils and was significantly higher under regenerative management compared to conventional practices in two out of four sites. SOC contents in perennial grasslands were up to 5% and always higher than in agricultural fields. Extractable organic carbon was similar in the two agricultural fields of the respective site, while grasslands diverged. Microbial biomass carbon was highest in grasslands at all sites and significantly higher in fields under regenerative agriculture compared to conventional agriculture at three out of four sites.Total nitrogen was highest in perennial grasslands at all sites, and similar in regenerative and conventional fields. The form of N however differed between soils under conventional and regenerative agriculture. Dissolved N, expressed per g total N was significantly higher or tended to be higher in conventional compared to regenerative agricultural fields. From this dissolved pool a higher proportion was in inorganic N forms that are more prone to leaching and gaseous loss compared to organic N forms. In soils from regenerative agricultural fields a higher proportion of the total N was found in the microbial biomass. This pool is considered to be highly dynamic, but also protected against losses. Less N in dissolved and inorganic form as well as a higher proportion of N in the microbial biomass indicates that the N cycle is more closed in soils managed regeneratively versus conventional.A greater importance of the microbial biomass could also have effects on soil C cycling. Higher microbial biomass is often related to increased carbon use efficiency, which in turn could indicate increased soil carbon sequestration. The already mentioned results will thus be discussed with further measurements of microbial respiration, growth and CUE.

The activities of soil microorganisms drive soil carbon (C) and nutrient cycling and therefore play an important role in local and global terrestrial C dynamics and nutrient cycles. Unfortunately, soil microbial activities have been defined mostly by measurements of heterotrophic respiration, potential enzyme activities, or net N processes. However, soil microbial activities comprise more than just catabolic processes such as respiration and N mineralization. Recently anabolic processes (biosynthesis and growth) and the partitioning between anabolic and catabolic processes in soil microbial metabolism have gained more attention as they control microbial soil organic matter formation. Understanding the controls on these processes allows an improved understanding of the key roles that soil microbes play in organic matter decomposition (catabolic processes) and soil organic matter sequestration (anabolic processes leading to growth, biomass and necromass formation), and their potential feedback to global change.Generally, there are two approaches to study the metabolism of soil microbial communities: First, position-specific isotope labeling is a tool that allows the tracing of 13C-atoms in organic molecules on their way through the network of metabolic pathways and second, metabolomics and fluxomics approaches can enable disentangling the highly complex metabolic networks of microbial communities, which however have rarely (metabolomics) or never (fluxomics) been applied to soils.In this study we developed a targeted soil metabolomics approach coupled to 13C isotope tracing (fluxomics), in which we extract, purify and measure a preselected set of key metabolites. Our aim was to cover the wide spectrum of soil microbial metabolic pathways based on the analysis of biomarker metabolites being unique to specific metabolic pathways such as glycolysis/gluconeogenesis (e.g. fructose 1,6-bisphosphate), the pentose phosphate pathway (ribose-5-phosphate), the citric acid cycle (α-ketoglutaric acid), purine and pyrimidine metabolism (UMP, AMP, allantoin), amino acid biosynthesis and degradation (10proteinogenic amino acids and their intermediates), the urea cycle (ornithine), amino sugar metabolism (N-Acetyl-D-Glucosamine and –muramic acid) and the shikimate pathway (shikimate). The minute concentrations of these primary metabolites are extracted from soils by 1 M KCl including 5 % chloroform, salts are removed by freeze-drying, methanol dissolution and cation-/anion-exchange chromatography and the metabolites and their isotopomers quantified by UPLC-Orbitrap mass spectrometry. To cover the wide range of metabolites, compound separations are performed by hydrophilic interaction chromatography (HILIC) for metabolites such as amino acids, (poly-)amines, nucleosides and nucleobases and by Ion chromatography (IC), to separate charged molecules like amino sugars, sugar phosphates and organic acids. Here we will show fluxomics results from a laboratory soil warming experiment where we added 13C-glucose to a temperate forest soil as a proof of concept.

Soil microbial physiology controls large fluxes of C to the atmosphere, thus, improving our ability to accurately quantify microbial physiology in soil is essential. However, current methods to determine microbial C metabolism require liquid water addition, which makes it practically impossible to measure microbial physiology in dry soil samples without stimulating microbial growth and respiration (namely, the “Birch effect”). We developed a new method based on in vivo 18O‐water vapor equilibration to minimize soil rewetting effects. This method allows the isotopic labeling of soil water without direct liquid water addition. This was compared to the main current method (direct 18O‐liquid water addition) in moist and air‐dry soils. We determined the time kinetics and calculated the average 18O enrichment of soil water over incubation time, which is necessary to calculate microbial growth from 18O incorporation in genomic DNA. We tested isotopic equilibration patterns in three natural and six artificially constructed soils covering a wide range of soil texture and soil organic matter content. We then measured microbial growth, respiration and carbon use efficiency (CUE) in three natural soils (either air‐dry or moist). The proposed 18O‐vapor equilibration method provided similar results as the current method of liquid 18O‐water addition when used for moist soils. However, when applied to air‐dry soils the liquid 18O‐water addition method overestimated growth by up to 250%, respiration by up to 500%, and underestimated CUE by up to 40%. We finally describe the new insights into biogeochemical cycling of C that the new method can help uncover, and we consider a range of questions regarding microbial physiology and its response to global change that can now be addressed., We developed a new method based on in vivo 18O‐water vapor equilibration to minimize soil rewetting effects. This method allows the isotopic labeling of soil water without any liquid water or dissolved substrate addition to the sample. When then applied it to dry soils to show that it allows proper estimation of microbial physiology, while the currently widely used method of liquid 18O‐water addition overestimated growth by up to 250%, respiration by up to 500%, and underestimated carbon use efficiency by up to 40%. We describe the new insights into biogeochemical cycling of C that the new method can help uncover.

Quantifying soil carbon dynamics is of utmost relevance in the context of global change because soils play an important role in land-atmosphere gas exchange. Our current understanding of both present and future carbon dynamics is limited because we fail to accurately represent soil processes across temporal and spatial scales, partly because of the paucity of data on the relative importance and hierarchical relationships between microbial, geochemical and climatic controls. Here, using observations from a 3,000-kyr-old soil chronosequence preserved in alluvial terrace deposits of the Merced River, California, we show how soil carbon dynamics are driven by the relationship between short-term biotic responses and long-term mineral weathering. We link temperature sensitivity of heterotrophic respiration to biogeochemical soil properties through their relationship with microbial activity and community composition. We found that soil mineralogy, and in particular changes in mineral reactivity and resulting nutrient availability, impacts the response of heterotrophic soil respiration to warming by altering carbon inputs, carbon stabilization, microbial community composition and extracellular enzyme activity. We demonstrate that biogeochemical alteration of the soil matrix (and not short-term warming) controls the composition of microbial communities and strategies to metabolize nutrients. More specifically, weathering first increases and then reduces nutrient availability and retention, as well as the potential of soils to stabilize carbon.

Despite decades of research progress, ecologists are still debating which pools and fluxes provide nitrogen (N) to plants and soil microbes across different ecosystems. Depolymerization of soil organic N is recognized as the rate-limiting step in the production of bioavailable N, and it is generally assumed that detrital N is the main source. However, in many mineral soils, detrital polymers constitute a minor fraction of total soil organic N. The majority of organic N is associated with clay-sized particles where physicochemical interactions may limit the accessibility of N-containing compounds. Although mineral-associated organic matter (MAOM) has historically been considered a critical, but relatively passive, reservoir of soil N, a growing body of research now points to the dynamic nature of mineral-organic associations and their potential for destabilization. Here we synthesize evidence from biogeoscience and soil ecology to demonstrate how MAOM is an important, yet overlooked, mediator of bioavailable N, especially in the rhizosphere. We highlight several biochemical strategies that enable plants and microbes to disrupt mineral-organic interactions and access MAOM. In particular, root-deposited low-molecular-weight exudates may enhance the mobilization and solubilization of MAOM, increasing its bioavailability. However, the competitive balance between the possible fates of N monomers—bound to mineral surfaces versus dissolved and available for assimilation—will depend on the specific interaction between mineral properties, soil solution, mineral-bound organic matter, and microbes. Building off our emerging understanding of MAOM as a source of bioavailable N, we propose a revision of the Schimel and Bennett (Ecology 85:591–602, 2004) model (which emphasizes N depolymerization), by incorporating MAOM as a potential proximal mediator of bioavailable N.

© 2018 Elsevier Ltd A substantial portion of soil organic matter (SOM) is of microbial origin. The efficiency with which soil microorganisms can convert their substrate carbon (C) into biomass, compared to how much is lost as respiration, thus co-determines the carbon storage potential of soils. Despite increasing insight into soil microbial C cycling, empirical measurements of microbial C processing across biomes and across soil horizons remain sparse. The theory of ecological stoichiometry predicts that microbial carbon use efficiency (CUE), i.e. growth over uptake of organic C, strongly depends on the relative availability of C and nutrients, particularly N, as microorganisms will either respire excess C or conserve C while mineralising excess nutrients. Microbial CUE is thus expected to increase from high to low latitudes and from topsoil to subsoil as the soil C:N and the stoichiometric imbalance between SOM and the microbial biomass decrease. To test these hypotheses, we collected soil samples from the organic topsoil, mineral topsoil, and mineral subsoil of seven sites along a 1500-km latitudinal transect in Western Siberia. As a proxy for CUE, we measured the microbial substrate use efficiency (SUE) of added substrates by incubating soil samples with a mixture of13C labelled sugars, amino sugars, amino acids, and organic acids and tracing13C into microbial biomass and released CO2. In addition to soil and microbial C:N stoichiometry, we also determined the potential extracellular enzyme activities of cellobiohydrolase (CBH) and phenol oxidase (POX) and used the CBH:POX ratio as an indicator of SOM substrate quality. We found an overall decrease of SUE with latitude, corresponding to a decrease in mean annual temperature, in mineral soil horizons. SUE decreased with decreasing stoichiometric imbalance in the organic and mineral topsoil, while a relationship of SUE with soil C:N was only found in the mineral topsoil. However, contrary to our hypothesis, SUE did not increase with soil depth and mineral subsoils displayed lower average SUE than mineral topsoils. Both within individual horizons and across all horizons SUE was strongly correlated with CBH:POX ratio as well as with climate variables. Since enzyme activities likely reflect the chemical properties of SOM, our results indicate that SOM quality exerts a stronger control on SUE than SOM stoichiometry, particularly in subsoils were SOM has been turned over repeatedly and there is little variation in SOM elemental ratios.

The occurrence of dark fixation of CO2 by heterotrophic microorganisms in soil is generally accepted, but its importance for microbial metabolism and soil organic carbon (C) sequestration is unknown, especially under C-limiting conditions. To fill this knowledge gap, we measured dark 13CO2 incorporation into soil organic matter and conducted a 13C-labelling experiment to follow the 13C incorporation into phospholipid fatty acids as microbial biomass markers across soil profiles of four tundra ecosystems in the northern circumpolar region, where net primary productivity and thus soil C inputs are low. We further determined the abundance of various carboxylase genes and identified their microbial origin with metagenomics. The microbial capacity for heterotrophic CO2 fixation was determined by measuring the abundance of carboxylase genes and the incorporation of 13C into soil C following the augmentation of bioavailable C sources. We demonstrate that dark CO2 fixation occurred ubiquitously in arctic tundra soils, with increasing importance in deeper soil horizons, presumably due to increasing C limitation with soil depth. Dark CO2 fixation accounted on average for 0.4, 1.0, 1.1, and 16% of net respiration in the organic, cryoturbated organic, mineral and permafrost horizons, respectively. Genes encoding anaplerotic enzymes of heterotrophic microorganisms comprised the majority of identified carboxylase genes. The genetic potential for dark CO2 fixation was spread over a broad taxonomic range. The results suggest important regulatory function of CO2 fixation in C limited conditions. The measurements were corroborated by modeling the long-term impact of dark CO2 fixation on soil organic matter. Our results suggest that increasing relative CO2 fixation rates in deeper soil horizons play an important role for soil internal C cycling and can, at least in part, explain the isotopic enrichment with soil depth.

Permafrost soils preserve huge amounts of organic carbon (OC) prone to decomposition under changing climatic conditions. However, knowledge on the composition of soil organic matter (OM) and its transformation and vulnerability to decomposition in these soils is scarce. We determined neutral sugars and lignin-derived phenols, released by trifluoroacetic acid (TFA) and CuO oxidation, respectively, within plants and soil density fractions from the active layer and the upper permafrost layer at three different tundra types (shrubby grass, shrubby tussock, shrubby lichen) in the Northeast Siberian Arctic. The heavy fraction (HF; > 1.6 g mL(-1)) was characterized by a larger enrichment of microbial sugars (hexoses vs. pentoses) and more pronotmced lignin degradation (acids vs. aldehydes) as compared to the light fraction (LF

Arctic permafrost soils contain large stocks of organic carbon (OC). Extensive cryogenic processes in these soils cause subduction of a significant part of OC-rich topsoil down into mineral soil through the process of cryoturbation. Currently, one-fourth of total permafrost OC is stored in subducted organic horizons. Predicted climate change is believed to reduce the amount of OC in permafrost soils as rising temperatures will increase decomposition of OC by soil microorganisms. To estimate the sensitivity of OC decomposition to soil temperature and oxygen levels we performed a 4-month incubation experiment in which we manipulated temperature (4–20 °C) and oxygen level of topsoil organic, subducted organic and mineral soil horizons. Carbon loss (CLOSS) was monitored and its potential biotic and abiotic drivers, including concentrations of available nutrients, microbial activity, biomass and stoichiometry, and extracellular oxidative and hydrolytic enzyme pools, were measured. We found that independently of the incubation temperature, CLOSS from subducted organic and mineral soil horizons was one to two orders of magnitude lower than in the organic topsoil horizon, both under aerobic and anaerobic conditions. This corresponds to the microbial biomass being lower by one to two orders of magnitude. We argue that enzymatic degradation of autochthonous subducted OC does not provide sufficient amounts of carbon and nutrients to sustain greater microbial biomass. The resident microbial biomass relies on allochthonous fluxes of nutrients, enzymes and carbon from the OC-rich topsoil. This results in a “negative priming effect”, which protects autochthonous subducted OC from decomposition at present. The vulnerability of subducted organic carbon in cryoturbated arctic soils under future climate conditions will largely depend on the amount of allochthonous carbon and nutrient fluxes from the topsoil.

In permafrost soils, the temperature regime and the resulting cryogenic processes are important determinants of the storage of organic carbon (OC) and its small-scale spatial variability. For cryoturbated soils, there is a lack of research assessing pedon-scale heterogeneity in OC stocks and the transformation of functionally different organic matter (OM) fractions, such as particulate and mineral-associated OM. Therefore, pedons of 28 Turbels were sampled in 5 m wide soil trenches across the Siberian Arctic to calculate OC and total nitrogen (TN) stocks based on digital profile mapping. Density fractionation of soil samples was performed to distinguish between particulate OM (light fraction, LF, < 1.6 g cm−3), mineral associated OM (heavy fraction, HF, > 1.6 g cm−3), and a mobilizable dissolved pool (mobilizable fraction, MoF). Across all investigated soil profiles, the total OC storage was 20.2 ± 8.0 kg m−2 (mean ± SD) to 100 cm soil depth. Fifty-four percent of this OC was located in the horizons of the active layer (annual summer thawing layer), showing evidence of cryoturbation, and another 35 % was present in the upper permafrost. The HF-OC dominated the overall OC stocks (55 %), followed by LF-OC (19 % in mineral and 13 % in organic horizons). During fractionation, approximately 13 % of the OC was released as MoF, which likely represents a readily bioavailable OM pool. Cryogenic activity in combination with cold and wet conditions was the principle mechanism through which large OC stocks were sequestered in the subsoil (16.4 ± 8.1 kg m−2; all mineral B, C, and permafrost horizons). Approximately 22 % of the subsoil OC stock can be attributed to LF material subducted by cryoturbation, whereas migration of soluble OM along freezing gradients appeared to be the principle source of the dominant HF (63 %) in the subsoil. Despite the unfavourable abiotic conditions, low C / N ratios and high δ13C values indicated substantial microbial OM transformation in the subsoil, but this was not reflected in altered LF and HF pool sizes. Partial least-squares regression analyses suggest that OC accumulates in the HF fraction due to co-precipitation with multivalent cations (Al, Fe) and association with poorly crystalline iron oxides and clay minerals. Our data show that, across all permafrost pedons, the mineral-associated OM represents the dominant OM fraction, suggesting that the HF-OC is the OM pool in permafrost soils on which changing soil conditions will have the largest impact. Russian Ministry of Education and Science/14.B25.31.0031 German Federal Ministry of Education and Research/03F0616A Evangelisches Studienwerk Villigst DFG

Summary Permafrost degradation may cause strong feedbacks of arctic ecosystems to global warming, but this will depend on if, and to what extent, organic matter (OM) is protected against biodegradation by mechanisms other than freezing and anoxia. Here, we report on the amount, chemical composition and bioavailability of particulate (POM) and mineral-associated OM (MOM) in permafrost soils of the East Siberian Arctic. The average total organic carbon (OC) stock across all soils was 24.0 ± 6.7 kg m−2 within 100 cm soil depth. Density fractionation (density cut-off 1.6 g cm−3) revealed that 54 ± 16% of the total soil OC and 64 ± 18% of OC in subsoil horizons was bound to minerals. As well as sorption of OM to clay-sized minerals (R2 = 0.80; P < 0.01), co-precipitation of OM with hydrolyzable metals may also transfer carbon into the mineral-bound fraction. Carbon:nitrogen ratios, stable carbon and nitrogen isotopes, 13C-NMR and X-ray photoelectron spectroscopy showed that OM is transformed in permafrost soils, which is a prerequisite for the formation of mineral-organic associations. Mineral-associated OM in deeper soil was enriched in 13C and 15N, and had narrow C:N and large alkyl C:(O-/N-alkyl C) ratios, indicating an advanced stage of decomposition. Despite being up to several thousands of years old, when incubated under favourable conditions (60% water-holding capacity, 15°C, adequate nutrients, 90 days), only 1.5–5% of the mineral-associated OC was released as CO2. In the topsoils, POM had the largest mineralization but was even less bioavailable than the MOM in subsoil horizons. Our results suggest that the formation of mineral-organic associations acts as an important additional factor in the stabilization of OM in permafrost soils. Although the majority of MOM was not prone to decomposition under favourable conditions, mineral-organic associations host a readily accessible carbon fraction, which may actively participate in ecosystem carbon exchange.

Climate change in Arctic ecosystems fosters permafrost thaw and makes massive amounts of ancient soil organic carbon (OC) available to microbial breakdown. However, fractions of the organic matter (OM) may be protected from rapid decomposition by their association with minerals. Little is known about the effects of mineral-organic associations (MOA) on the microbial accessibility of OM in permafrost soils and it is not clear which factors control its temperature sensitivity. In order to investigate if and how permafrost soil OC turnover is affected by mineral controls, the heavy fraction (HF) representing mostly MOA was obtained by density fractionation from 27 permafrost soil profiles of the Siberian Arctic. In parallel laboratory incubations, the unfractionated soils (bulk) and their HF were comparatively incubated for 175 days at 5 and 15°C. The HF was equivalent to 70 ± 9% of the bulk CO2 respiration as compared to a share of 63 ± 1% of bulk OC that was stored in the HF. Significant reduction of OC mineralization was found in all treatments with increasing OC content of the HF (HF-OC), clay-size minerals and Fe or Al oxyhydroxides. Temperature sensitivity (Q10) decreased with increasing soil depth from 2.4 to 1.4 in the bulk soil and from 2.9 to 1.5 in the HF. A concurrent increase in the metal-to-HF-OC ratios with soil depth suggests a stronger bonding of OM to minerals in the subsoil. There, the younger 14C signature in CO2 than that of the OC indicates a preferential decomposition of the more recent OM and the existence of a MOA fraction with limited access of OM to decomposers. These results indicate strong mineral controls on the decomposability of OM after permafrost thaw and on its temperature sensitivity. Thus, we here provide evidence that OM temperature sensitivity can be attenuated by MOA in permafrost soils.

Analysis of disturbance effects on multiple microbial processes elucidates response of biogeochemical cycling to climate extremes., Predicted changes in the intensity and frequency of climate extremes urge a better mechanistic understanding of the stress response of microbially mediated carbon (C) and nutrient cycling processes. We analyzed the resistance and resilience of microbial C, nitrogen (N), and phosphorus (P) cycling processes and microbial community composition in decomposing plant litter to transient, but severe, temperature disturbances, namely, freeze-thaw and heat. Disturbances led temporarily to a more rapid cycling of C and N but caused a down-regulation of P cycling. In contrast to the fast recovery of the initially stimulated C and N processes, we found a slow recovery of P mineralization rates, which was not accompanied by significant changes in community composition. The functional and structural responses to the two distinct temperature disturbances were markedly similar, suggesting that direct negative physical effects and costs associated with the stress response were comparable. Moreover, the stress response of extracellular enzyme activities, but not that of intracellular microbial processes (for example, respiration or N mineralization), was dependent on the nutrient content of the resource through its effect on microbial physiology and community composition. Our laboratory study provides novel insights into the mechanisms of microbial functional stress responses that can serve as a basis for field studies and, in particular, illustrates the need for a closer integration of microbial C-N-P interactions into climate extremes research.

Cryoturbation, the burial of topsoil material into deeper soil horizons by repeated freeze–thaw events, is an important storage mechanism for soil organic matter (SOM) in permafrost-affected soils. Besides abiotic conditions, microbial community structure and the accessibility of SOM to the decomposer community are hypothesized to control SOM decomposition and thus have a crucial role in SOM accumulation in buried soils. We surveyed the microbial community structure in cryoturbated soils from nine soil profiles in the northeastern Siberian tundra using high-throughput sequencing and quantification of bacterial, archaeal and fungal marker genes. We found that bacterial abundances in buried topsoils were as high as in unburied topsoils. In contrast, fungal abundances decreased with depth and were significantly lower in buried than in unburied topsoils resulting in remarkably low fungal to bacterial ratios in buried topsoils. Fungal community profiling revealed an associated decrease in presumably ectomycorrhizal (ECM) fungi. The abiotic conditions (low to subzero temperatures, anoxia) and the reduced abundance of fungi likely provide a niche for bacterial, facultative anaerobic decomposers of SOM such as members of the Actinobacteria, which were found in significantly higher relative abundances in buried than in unburied topsoils. Our study expands the knowledge on the microbial community structure in soils of Northern latitude permafrost regions, and attributes the delayed decomposition of SOM in buried soils to specific microbial taxa, and particularly to a decrease in abundance and activity of ECM fungi, and to the extent to which bacterial decomposers are able to act as their functional substitutes.

Thermal adaptations of soil microorganisms could mitigate or facilitate global warming effects on soil organic matter (SOM) decomposition and soil CO2 efflux. We incubated soil from warmed and control subplots of a forest soil warming experiment to assess whether 9 years of soil warming affected the rates and the temperature sensitivity of the soil CO2 efflux, extracellular enzyme activities, microbial efficiency, and gross N mineralization. Mineral soil (0–10 cm depth) was incubated at temperatures ranging from 3 to 23 °C. No adaptations to long‐term warming were observed regarding the heterotrophic soil CO2 efflux (R 10 warmed: 2.31 ± 0.15 μmol m−2 s−1, control: 2.34 ± 0.29 μmol m−2 s−1; Q 10 warmed: 2.45 ± 0.06, control: 2.45 ± 0.04). Potential enzyme activities increased with incubation temperature, but the temperature sensitivity of the enzymes did not differ between the warmed and the control soils. The ratio of C : N acquiring enzyme activities was significantly higher in the warmed soil. Microbial biomass‐specific respiration rates increased with incubation temperature, but the rates and the temperature sensitivity (Q 10 warmed: 2.54 ± 0.23, control 2.75 ± 0.17) did not differ between warmed and control soils. Microbial substrate use efficiency (SUE) declined with increasing incubation temperature in both, warmed and control, soils. SUE and its temperature sensitivity (Q 10 warmed: 0.84 ± 0.03, control: 0.88 ± 0.01) did not differ between warmed and control soils either. Gross N mineralization was invariant to incubation temperature and was not affected by long‐term soil warming. Our results indicate that thermal adaptations of the microbial decomposer community are unlikely to occur in C‐rich calcareous temperate forest soils.

The storage of soil samples for PLFA analysis can lead to shifts in the microbial community composition. We show here that conserving samples in RNAlater, which is already widely used to store samples for DNA and RNA analysis, proved to be as sufficient as freezing at −20 °C and preferable over storage at 4 °C for temperate mountain grassland soil. The total amount of extracted PLFAs was not changed by any storage treatment. Storage at 4 °C led to an alteration of seven out of thirty individual biomarkers, while freezing and storage in RNAlater caused changes in the amount of fungal biomarkers but had no effect on any other microbial group. We therefore suggest that RNAlater could be used to preserve soil samples for PLFA analysis when immediate extraction or freezing of samples is not possible, for example during sampling campaigns in remote areas or during transport and shipping., Highlights ► We tested storage of soils in RNAlater for analysis of phospholipid fatty acids. ► Storage in RNAlater proofed to be comparable to freezing at −20 °C. ► Freezing and RNAlater led to reduced fungal biomarkers compared to controls. ► Storage in RNAlater is suggested as a novel approach for sampling at remote areas.

Soil horizons below 30 cm depth contain about 60% of the organic carbon stored in soils. Although insight into the physical and chemical stabilization of soil organic matter (SOM) and into microbial community composition in these horizons is being gained, information on microbial functions of subsoil microbial communities and on associated microbially-mediated processes remains sparse. To identify possible controls on enzyme patterns, we correlated enzyme patterns with biotic and abiotic soil parameters, as well as with microbial community composition, estimated using phospholipid fatty acid profiles. Enzyme patterns (i.e. distance-matrixes calculated from these enzyme activities) were calculated from the activities of six extracellular enzymes (cellobiohydrolase, leucine-amino-peptidase, N-acetylglucosaminidase, chitotriosidase, phosphatase and phenoloxidase), which had been measured in soil samples from organic topsoil horizons, mineral topsoil horizons, and mineral subsoil horizons from seven ecosystems along a 1500 km latitudinal transect in Western Siberia. We found that hydrolytic enzyme activities decreased rapidly with depth, whereas oxidative enzyme activities in mineral horizons were as high as, or higher than in organic topsoil horizons. Enzyme patterns varied more strongly between ecosystems in mineral subsoil horizons than in organic topsoils. The enzyme patterns in topsoil horizons were correlated with SOM content (i.e., C and N content) and microbial community composition. In contrast, the enzyme patterns in mineral subsoil horizons were related to water content, soil pH and microbial community composition. The lack of correlation between enzyme patterns and SOM quantity in the mineral subsoils suggests that SOM chemistry, spatial separation or physical stabilization of SOM rather than SOM content might determine substrate availability for enzymatic breakdown. The correlation of microbial community composition and enzyme patterns in all horizons, suggests that microbial community composition shapes enzyme patterns and might act as a modifier for the usual dependency of decomposition rates on SOM content or C/N ratios., Highlights • Enzyme patterns differed more strongly between horizons than between ecosystems. • Enzyme patterns in subsoils vary more strongly than enzyme patterns in topsoils. • Enzyme patterns in topsoil are related to SOM content and mic. community composition. • Enzyme patterns in mineral subsoil were not related to SOM content or SOM C/N.

Permafrost-affected soils in the Northern latitudes store huge amounts of organic carbon (OC) that is prone to microbial degradation and subsequent release of greenhouse gasses to the atmosphere. In Greenland, the consequences of permafrost thaw have only recently been addressed, and predictions on its impact on the carbon budget are thus still highly uncertain. However, the fate of OC is not only determined by abiotic factors, but closely tied to microbial activity. We investigated eight soil profiles in northeast Greenland comprising two sites with typical tundra vegetation and one wet fen site. We assessed microbial community structure and diversity (SSU rRNA gene tag sequencing, quantification of bacteria, archaea and fungi), and measured hydrolytic and oxidative enzyme activities. Sampling site and thus abiotic factors had a significant impact on microbial community structure, diversity and activity, the wet fen site exhibiting higher potential enzyme activities and presumably being a hot spot for anaerobic degradation processes such as fermentation and methanogenesis. Lowest fungal to bacterial ratios were found in topsoils that had been relocated by cryoturbation (“buried topsoils”), resulting from a decrease in fungal abundance compared to recent (“unburied”) topsoils. Actinobacteria (in particular Intrasporangiaceae) accounted for a major fraction of the microbial community in buried topsoils, but were only of minor abundance in all other soil horizons. It was indicated that the distribution pattern of Actinobacteria and a variety of other bacterial classes was related to the activity of phenol oxidases and peroxidases supporting the hypothesis that bacteria might resume the role of fungi in oxidative enzyme production and degradation of phenolic and other complex substrates in these soils. Our study sheds light on the highly diverse, but poorly-studied communities in permafrost-affected soils in Greenland and their role in OC degradation.

Summary The proportion of carbon allocated to wood production is an important determinant of the carbon sink strength of global forest ecosystems. Understanding the mechanisms controlling wood production and its responses to environmental drivers is essential for parameterization of global vegetation models and to accurately predict future responses of tropical forests in terms of carbon sequestration. Here, we synthesize data from 105 pantropical old-growth rainforests to investigate environmental controls on the partitioning of net primary production to wood production (%WP) using structural equation modeling. Our results reveal that %WP is governed by two independent pathways of direct and indirect environmental controls. While temperature and soil phosphorus availability indirectly affected %WP via increasing productivity, precipitation and dry season length both directly increased %WP via tradeoffs along the plant economics spectrum. We provide new insights into the mechanisms driving %WP, allowing us to conclude that projected climate change could enhance %WP in less productive tropical forests, thus increasing carbon sequestration in montane forests, but adversely affecting lowland forests.

Turbic Cryosols (permafrost soils characterized by cryoturbation, i.e., by mixing of soil layers due to freezing and thawing) are widespread across the Arctic, and contain large amounts of poorly decomposed organic material buried in the subsoil. This cryoturbated organic matter exhibits retarded decomposition compared to organic material in the topsoil. Since soil organic matter (SOM) decomposition is known to be tightly linked to N availability, we investigated N transformation rates in different soil horizons of three tundra sites in north-eastern Siberia and Greenland. We measured gross rates of protein depolymerization, N mineralization (ammonification) and nitrification, as well as microbial uptake of amino acids and NH4+ using an array of 15N pool dilution approaches. We found that all sites and horizons were characterized by low N availability, as indicated by low N mineralization compared to protein depolymerization rates (with gross N mineralization accounting on average for 14% of gross protein depolymerization). The proportion of organic N mineralized was significantly higher at the Greenland than at the Siberian sites, suggesting differences in N limitation. The proportion of organic N mineralized, however, did not differ significantly between soil horizons, pointing to a similar N demand of the microbial community of each horizon. In contrast, absolute N transformation rates were significantly lower in cryoturbated than in organic horizons, with cryoturbated horizons reaching not more than 32% of the transformation rates in organic horizons. Our results thus indicate a deceleration of the entire N cycle in cryoturbated soil horizons, especially strongly reduced rates of protein depolymerization (16% of organic horizons) which is considered the rate-limiting step in soil N cycling., Highlights • N turnover in cryoturbated Arctic soils measured by isotope pool dilution assays. • Protein depolymerization strongly exceeds NH4+ and NO3− production rates. • Microbial N use efficiency indicates N limitation in Siberian sampling sites. • Deceleration of microbial N transformation rates by cryoturbation.

Summary • Plant seasonal cycles alter carbon (C) and nitrogen (N) availability for soil microbes, which may affect microbial community composition and thus feed back on microbial decomposition of soil organic material and plant N availability. The temporal dynamics of these plant–soil interactions are, however, unclear. • Here, we experimentally manipulated the C and N availability in a beech forest through N fertilization or tree girdling and conducted a detailed analysis of the seasonal pattern of microbial community composition and decomposition processes over 2 yr. • We found a strong relationship between microbial community composition and enzyme activities over the seasonal course. Phenoloxidase and peroxidase activities were highest during late summer, whereas cellulase and protease peaked in late autumn. Girdling, and thus loss of mycorrhiza, resulted in an increase in soil organic matter-degrading enzymes and a decrease in cellulase and protease activity. • Temporal changes in enzyme activities suggest a switch of the main substrate for decomposition between summer (soil organic matter) and autumn (plant litter). Our results indicate that ectomycorrhizal fungi are possibly involved in autumn cellulase and protease activity. Our study shows that, through belowground C allocation, trees significantly alter soil microbial communities, which may affect seasonal patterns of decomposition processes.

Soil organic matter (SOM) is important for soil fertility and climate change mitigation. Agricultural management - including soil amendments - can improve soil fertility and contribute to climate change mitigation by stabilising carbon in soils. This calls for cost-effective parameters to assess the influence of management practices on SOM. The current study aimed at understanding how sensitive the parameters active/permanganate oxidisable carbon (AC) and nitrogen mineralisation potential (NMP) react to different agricultural management practices compared to total organic carbon (TOC) and total nitrogen (Nt). We aimed to gain a better understanding of SOM processes, mainly regarding depth distribution and seasonality of SOM dynamics using AC and NMP.Data were obtained in five Austrian long-term field experiments (LTEs) testing four management practices: i) tillage, ii) compost application, iii) crop residue management, and iv) mineral fertilisation.AC was specifically sensitive in detecting the effect of tillage treatment at different soil depths. NMP differentiated between all different tillage treatments in the top soil layer, it showed the temporal dynamics between the years in the compost LTE, and it was identified as an early detection property in the crop residue LTE. Both AC and NMP detected short-term fluctuations better than TOC and Nt over the course of two years in the crop residue LTE. Thus, we suggest that AC and NMP are two valuable soil biochemical parameters providing more detailed information on C and N dynamics regarding depth distribution and seasonal dynamics and react more sensitively to different agricultural management practices compared to TOC and Nt. They should be integrated in monitoring agricultural LTEs and in field analyses conducted by farmers. However, when evaluating results of long-term carbon storage, their sensitivity towards annual fluctuations should be taken into account.