Search

Terrestrial ecosystems remove about 30 per cent of the carbon dioxide (CO2) emitted by human activities each year1, yet the persistence of this carbon sink depends partly on how plant biomass and soil organic carbon (SOC) stocks respond to future increases in atmospheric CO2 (refs. 2,3). Although plant biomass often increases in elevated CO2 (eCO2) experiments4–6, SOC has been observed to increase, remain unchanged or even decline7. The mechanisms that drive this variation across experiments remain poorly understood, creating uncertainty in climate projections8,9. Here we synthesized data from 108 eCO2 experiments and found that the effect of eCO2 on SOC stocks is best explained by a negative relationship with plant biomass: when plant biomass is strongly stimulated by eCO2, SOC storage declines; conversely, when biomass is weakly stimulated, SOC storage increases. This trade-off appears to be related to plant nutrient acquisition, in which plants increase their biomass by mining the soil for nutrients, which decreases SOC storage. We found that, overall, SOC stocks increase with eCO2 in grasslands (8 ± 2 per cent) but not in forests (0 ± 2 per cent), even though plant biomass in grasslands increase less (9 ± 3 per cent) than in forests (23 ± 2 per cent). Ecosystem models do not reproduce this trade-off, which implies that projections of SOC may need to be revised. A synthesis of elevated carbon dioxide experiments reveals that when plant biomass is strongly stimulated by elevated carbon dioxide levels, soil carbon storage declines, and where biomass is weakly stimulated, soil carbon accumulates.

Through agriculture and industry, humans are increasing the deposition and availability of nitrogen (N) in ecosystems worldwide. Carbon (C) isotope tracers provide useful insights into soil C dynamics, as they allow to study soil C pools of different ages. We evaluated to what extent N enrichment affects soil C dynamics in experiments that applied C isotope tracers. Using meta-analysis, we synthesized data from 35 published papers. We made a distinction between “new C” and “old C” stocks, i.e., soil C derived from plant C input since the start of the isotopic enrichment, or unlabeled, pre-existing soil C. Averaged across studies, N addition increased new soil C stocks (+30.3%), total soil C stocks (+6.1%) and soil C input proxies (+30.7%). Although N addition had no overall, average, effect on old soil C stocks and old soil C respiration, old soil C stocks increased with the amount of N added and respiration of old soil C declined. Nitrogen-induced effects on new soil C and soil C input both decreased with the amount of extraneous N added in control treatments. Although our findings require additional confirmation from long-term field experiments, our analysis provides isotopic evidence that N addition stimulates soil C storage both by increasing soil C input and (at high N rates) by decreasing decomposition of old soil C. Furthermore, we demonstrate that the widely reported saturating response of plant growth to N enrichment also applies to new soil C storage.

Elevated atmospheric CO2 (eCO2 ) generally increases carbon input in rice paddy soils and stimulates the growth of methane-producing microorganisms. Therefore, eCO2 is widely expected to increase methane (CH4 ) emissions from rice agriculture, a major source of anthropogenic CH4 . Agricultural practices strongly affect CH4 emissions from rice paddies as well, but whether these practices modulate effects of eCO2 is unclear. Here we show, by combining a series of experiments and meta-analyses, that whereas eCO2 strongly increased CH4 emissions from paddies without straw incorporation, it tended to reduce CH4 emissions from paddy soils with straw incorporation. Our experiments also identified the microbial processes underlying these results: eCO2 increased methane-consuming microorganisms more strongly in soils with straw incorporation than in soils without straw, with the opposite pattern for methane-producing microorganisms. Accounting for the interaction between CO2 and straw management, we estimate that eCO2 increases global CH4 emissions from rice paddies by 3.7%, an order of magnitude lower than previous estimates. Our results suggest that the effect of eCO2 on CH4 emissions from rice paddies is smaller than previously thought and underline the need for judicious agricultural management to curb future CH4 emissions.

RNA is considered to be a short-lived molecule, indicative of cellular metabolic activity, whereas DNA is thought to turn over more slowly because living cells do not always grow and divide. To explore differences in the rates of synthesis of these nucleic acids, we used H218O quantitative stable isotope probing (qSIP) to measure the incorporation of 18O into 16S rRNA, the 16S rDNA, amoA mRNA and the amoA gene of soil Thaumarchaeota. Incorporation of 18O into the thaumarchaeal amoA mRNA pool was faster than into the 16S rRNA pool, suggesting that Thaumarchaea were metabolically active while using rRNA molecules that were likely synthetized prior to H218O addition. Assimilation rates of 18O into 16S rDNA and amoA genes were similar, which was expected because both genes are present in the same thaumarchaeal genome. The Thaumarchaea had significantly higher rRNA to rDNA ratios than bacteria, though the 18O isotopic signature of thaumarchaeal rRNA was lower than that of bacterial rRNA, further suggesting preservation of old non-labeled rRNA. Through qSIP of soil with H218O, we showed that 18O incorporation into thaumarchaeal nucleic acids was generally low, indicating slower turnover rates compared to bacteria, and potentially suggesting thaumarchaeal capability for preservation and efficient reuse of biomolecules.

Plants may slow global warming through enhanced growth, because increased levels of photosynthesis stimulate the land carbon (C) sink. However, the key drivers determining responses of plants to warming remain unclear, causing uncertainty in climate projections. Using meta- analysis, we show that the effect of experimental warming on plant biomass is best explained by soil nitrogen (N) availability. Warming-induced changes in total, aboveground and belowground biomass all positively correlated with soil C:N ratio, an indicator of soil N availability. In factorial N × warming experiments, warming increased plant biomass more strongly under low N than under high N availability. Together, these results suggest that warming stimulates plant C storage most strongly in ecosystems where N limits plant growth. Thus, incorporating the soil N status of ecosystems into Earth system models may improve predictions of future carbon-climate feedbacks.

A large portion of activity in soil microbial communities occurs in short time frames in response to an increase in C availability, affecting the biogeochemical cycling of nitrogen. These changes are of particular importance as nitrogen represents both a limiting nutrient for terrestrial plants as well as a potential pollutant., Episodic inputs of labile carbon (C) to soil can rapidly stimulate nitrogen (N) immobilization by soil microorganisms. However, the transcriptional patterns that underlie this process remain unclear. In order to better understand the regulation of N cycling in soil microbial communities, we conducted a 48-h laboratory incubation with agricultural soil where we stimulated the uptake of inorganic N by amending the soil with glucose. We analyzed the metagenome and metatranscriptome of the microbial communities at four time points that corresponded with changes in N availability. The relative abundances of genes remained largely unchanged throughout the incubation. In contrast, glucose addition rapidly increased the transcription of genes encoding ammonium and nitrate transporters, enzymes responsible for N assimilation into biomass, and genes associated with the N regulatory network. This upregulation coincided with an increase in transcripts associated with glucose breakdown and oxoglutarate production, demonstrating a connection between C and N metabolism. When concentrations of ammonium were low, we observed a transient upregulation of genes associated with the nitrogen-fixing enzyme nitrogenase. Transcripts for nitrification and denitrification were downregulated throughout the incubation, suggesting that dissimilatory transformations of N may be suppressed in response to labile C inputs in these soils. These results demonstrate that soil microbial communities can respond rapidly to changes in C availability by drastically altering the transcription of N cycling genes. IMPORTANCE A large portion of activity in soil microbial communities occurs in short time frames in response to an increase in C availability, affecting the biogeochemical cycling of nitrogen. These changes are of particular importance as nitrogen represents both a limiting nutrient for terrestrial plants as well as a potential pollutant. However, we lack a full understanding of the short-term effects of labile carbon inputs on the metabolism of microbes living in soil. Here, we found that soil microbial communities responded to labile carbon addition by rapidly transcribing genes encoding proteins and enzymes responsible for inorganic nitrogen acquisition, including nitrogen fixation. This work demonstrates that soil microbial communities respond within hours to carbon inputs through altered gene expression. These insights are essential for an improved understanding of the microbial processes governing soil organic matter production, decomposition, and nutrient cycling in natural and agricultural ecosystems.

Climate warming is known to impact ecosystem composition and functioning. However, it remains largely unclear how soil microbial communities respond to long-term, moderate warming. In this study, we used Illumina sequencing and microarrays (GeoChip 5.0) to analyze taxonomic and functional gene compositions of the soil microbial community after 14 years of warming (at 0.8–1.0 °C for 10 years and then 1.5–2.0 °C for 4 years) in a Californian grassland. Long-term warming had no detectable effect on the taxonomic composition of soil bacterial community, nor on any plant or abiotic soil variables. In contrast, functional gene compositions differed between warming and control for bacterial, archaeal, and fungal communities. Functional genes associated with labile carbon (C) degradation increased in relative abundance in the warming treatment, whereas those associated with recalcitrant C degradation decreased. A number of functional genes associated with nitrogen (N) cycling (e.g., denitrifying genes encoding nitrate-, nitrite-, and nitrous oxidereductases) decreased, whereas nifH gene encoding nitrogenase increased in the warming treatment. These results suggest that microbial functional potentials are more sensitive to long-term moderate warming than the taxonomic composition of microbial community.

International audience; Atmospheric carbon dioxide concentration ([CO 2 ]) is increasing, which increases leaf-scale photosynthesis and intrinsic water-use efficiency. These direct responses have the potential to increase plant growth, vegetation biomass, and soil organic matter; transferring carbon from the atmosphere into terrestrial ecosystems (a carbon sink). A substantial global terrestrial carbon sink would slow the rate of [CO 2] increase and thus climate change. However, ecosystem CO2 responses are complex or confounded by concurrent changes in multiple agents of global change and evidence for a [CO 2]-driven terrestrial carbon sink can appear contradictory. Here we synthesize theory and broad, multidisciplinary evidence for the effects of increasing [CO 2] (iCO 2) on the global terrestrial carbon sink. Evidence suggests a substantial increase in global photosynthesis since pre-industrial times. Established theory, supported by experiments, indicates that iCO 2 is likely responsible for about half of the increase. Global carbon budgeting, atmospheric data, and forest inventories indicate a historical carbon sink, and these apparent iCO 2 responses are high in comparison to experiments and predictions from theory. Plantmortality and soil carbon iCO 2 responses are highly uncertain. In conclusion, a range of evidence supports a positive terrestrial carbon sink in response to iCO2 , albeit with uncertain magnitude and strong suggestion of a role for additional agents of global change.

Fungi relieve nitrogen limitation Rising concentrations of atmospheric CO 2 stimulate plant growth; an effect that could reduce the pace of anthropogenic climate change. But plants also need nitrogen for growth. So far, experimental nitrogen addition has had equivocal effects on the magnitude of CO 2 fertilization. Terrer et al. explain that the impact of nitrogen on plant growth depends on the relationship between nitrogen availability and symbioses with mycorrhizal soil fungi. Only plants with ectomycorrhizal fungi associated with their roots can overcome nitrogen limitation. Science , this issue p. 72

Nutrient amendment diminished bacterial functional diversity, consolidating carbon flow through fewer bacterial taxa. Here, we show strong differences in the bacterial taxa responsible for respiration from four ecosystems, indicating the potential for taxon-specific control over soil carbon cycling. Trends in functional diversity, defined as the richness of bacteria contributing to carbon flux and their equitability of carbon use, paralleled trends in taxonomic diversity although functional diversity was lower overall. Among genera common to all ecosystems, Bradyrhizobium, the Acidobacteria genus RB41, and Streptomyces together composed 45–57% of carbon flow through bacterial productivity and respiration. Bacteria that utilized the most carbon amendment (glucose) were also those that utilized the most native soil carbon, suggesting that the behavior of key soil taxa may influence carbon balance. Mapping carbon flow through different microbial taxa as demonstrated here is crucial in developing taxon-sensitive soil carbon models that may reduce the uncertainty in climate change projections., The fate of soil carbon depends on microbial processes, but whether different microbial taxa have individualistic effects on carbon fluxes is unknown. Here the authors use 16 S amplicon sequencing and stable isotopes to show how taxonomic differences influence bacterial respiration and carbon cycling across four ecosystems.

Increased human‐derived nitrogen (N) deposition to terrestrial ecosystems has resulted in widespread phosphorus (P) limitation of net primary productivity. However, it remains unclear if and how N‐induced P limitation varies over time. Soil extracellular phosphatases catalyze the hydrolysis of P from soil organic matter, an important adaptive mechanism for ecosystems to cope with N‐induced P limitation. Here we show, using a meta‐analysis of 140 studies and 668 observations worldwide, that N stimulation of soil phosphatase activity diminishes over time. Whereas short‐term N loading (≤5 years) significantly increased soil phosphatase activity by 28%, long‐term N loading had no significant effect. Nitrogen loading did not affect soil available P and total P content in either short‐ or long‐term studies. Together, these results suggest that N‐induced P limitation in ecosystems is alleviated in the long‐term through the initial stimulation of soil phosphatase activity, thereby securing P supply to support plant growth. Our results suggest that increases in terrestrial carbon uptake due to ongoing anthropogenic N loading may be greater than previously thought. This study was funded by Aarhus University Centre for Circular Bioeconomy, Aarhus University Research Foundation AUFF Starting Grants (AUFF-E-2019-7-1), and Marie Skłodowska-Curie Individual Fellowship H2020-MSCA-IF-2018 (no. 839806). Ji Chen acknowledges funding support from the National Natural Science Foundation of China (41701292) and China Postdoctoral Science Foundation (2017M610647, 2018T111091) when constructing the databases. César Terrer was supported by a Lawrence Fellow award through Lawrence Livermore National Laboratory (LLNL). This work was performed under the auspices of the U.S. Department of Energy by LLNL under contract DEAC52-07NA27344 and was supported by the LLNL-LDRD Program under Project No. 20-ERD-055. Fernando T. Maestre was supported by the European Research Council (ERC Grant agreement 647038 [BIODESERT]) and Generalitat Valenciana (CIDEGENT/2018/041).

Climate warming affects soil carbon (C) dynamics, with possible serious consequences for soil C stocks and atmospheric CO2 concentrations. However, the mechanisms underlying changes in soil C storage are not well understood, hampering long-term predictions of climate C-feedbacks. The activity of the extracellular enzymes ligninase and cellulase can be used to track changes in the predominant C sources of soil microbes and can thus provide mechanistic insights into soil C loss pathways. Here we show, using meta-analysis, that reductions in soil C stocks with warming are associated with increased ratios of ligninase to cellulase activity. Furthermore, whereas long-term (≥5 years) warming reduced the soil recalcitrant C pool by 14%, short-term warming had no significant effect. Together, these results suggest that warming stimulates microbial utilization of recalcitrant C pools, possibly exacerbating long-term climate-C feedbacks.

Soil contains the largest terrestrial pool of carbon (C), but how this pool will be affected by global change remains unknown. Warmer temperatures generally increase soil respiration, while additional C inputs from plants to soil can increase or decrease soil C decomposition rates through a phenomenon known as priming. Priming occurs when soil organic matter (SOM) decomposition rates change in response to a fresh substrate, though the mechanisms underlying priming are poorly understood. Here, we measured priming in four ecosystems during a seven-week incubation with weekly glucose additions. Soil was collected from field warming experiments in the four ecosystems, so our experiment assessed the influence of long-term warming on priming. All treatments exhibited negative priming (reduced SOM decomposition) after the first substrate pulse. Subsequent substrate pulses elicited variable responses, and the effect of long-term warming on priming was ecosystem-dependent. Priming was correlated with changes in soil C and N in response to warming: ecosystems that lost soil C and N over nine years of experimental warming exhibited low rates of priming (decreased SOM decomposition), while ecosystems that gained soil C and N in response to warming had high priming. Consequently, priming may accelerate C losses in ecosystems that exhibit warming-induced C increases, and vice versa, thus partially buffering soil C content against change.

Microbes decompose soil organic matter (SOM), yet it is unclear how substrate inputs (i.e., stoichiometry) directly mediate microbial activities and community dynamics. We hypothesized that C+N input has the largest effect on microbial respiration and community structure, followed by C input and N input. Soils were collected from four ecosystems (grassland, pinon-juniper, ponderosa pine, mixed conifer) and amended with NH4NO3 (N only; 100 μg g−1 wk−1), 13C-glucose (C only; 1000 μg g−1 wk−1), or C+N in a five-week laboratory incubation. We found that C+N input induced the greatest total respiration while C input induced the greatest SOM-derived respiration (i.e., priming effect) across ecosystems. Shifts in community composition were the largest with C+N input, followed by C input, and showed little response to N input. C only and C+N inputs increased both of the relative and absolute abundances of Actinobacteria and Proteobacteria (α, β, γ), but reduced the relative abundances of Verrucomicrobia and δ-Proteobacteria. C+N input increased the relative abundances of Bacillales, Rhizobiales, Burkholderiales and of 9 families, and reduced the relative abundances of Myxococcales and of 12 families, but showed little effect on the absolute abundances of these bacterial taxa. N input reduced the absolute abundances of Actinobacteria, Proteobacteria, and Verrucomicrobia but did not affect their relative abundances in the mixed conifer soil; by contrast, N input reduced relative abundances of δ-Proteobacteria and increased the relative abundances of γ-Proteobacteria but did not affect their absolute abundances in the ponderosa pine soil. We also found that substrate inputs were the main driver of SOM decomposition, microbial respiration and diversity, while soil ecosystem was the main driver of community composition and abundances of most bacterial phyla. Our work suggests that substrate stoichiometry has predictable effects on soil C cycling, microbial diversity and community composition, but has variable effects on microbial abundances, and that incorporating bacterial gene copies in abundance calculations can help more accurately estimate microbial responses across taxonomic levels and ecosystems.

Even though microbial communities can be more effective at degrading xenobiotics than cultured micro-organisms, yet little is known about the microbial strategies that underpin xenobiotic biodegradation by microbial communities. Here, we employ metagenomic community sequencing to explore the mechanisms that drive the development of 49 xenobiotic-degrading microbial communities, which were enriched from 7 contaminated soils or sediments with a range of xenobiotic compounds. We show that multiple microbial strategies likely drive the development of xenobiotic degrading communities, notably (i) presence of genes encoding catabolic enzymes to degrade xenobiotics; (ii) presence of genes encoding efflux pumps; (iii) auxiliary catabolic genes on plasmids; and (iv) positive interactions dominate microbial communities with efficient degradation. Overall, the integrated analyses of microbial ecological strategies advance our understanding of microbial processes driving the biodegradation of xenobiotics and promote the design of bioremediation systems.

Rising levels of atmospheric CO2 frequently stimulate plant inputs to soil, but the consequences of these changes for soil carbon (C) dynamics are poorly understood. Plant-derived inputs can accumulate in the soil and become part of the soil C pool (“new soil C”), or accelerate losses of pre-existing (“old”) soil C. The dynamics of the new and old pools will likely differ and alter the long-term fate of soil C, but these separate pools, which can be distinguished through isotopic labeling, have not been considered in past syntheses. Using meta-analysis, we found that while elevated CO2 (ranging from 550 to 800 parts per million by volume) stimulates the accumulation of new soil C in the short term (

Fertilizer inputs affect plant uptake of native soil nitrogen (N), yet the underlying mechanisms remain elusive. To increase mechanistic insight into this phenomenon, we evaluated the effect of fertilizer addition on mineralization (in the absence of plants) and plant uptake of native soil N. We synthesized 43 isotope tracer (15N) studies and estimated the effects of fertilizer addition using meta-analysis. We found that organic fertilizer tended to reduce native soil N mineralization (−99 kg ha−1 year−1; p = 0.09) while inorganic fertilizer tended to increase N priming (58 kg ha−1 year−1; p = 0.17). In contrast, both organic and inorganic fertilizers significantly increased plant uptake of native soil N (179 and 107 kg ha−1 year−1). Organic fertilizer had greater effect on plant uptake than on mineralization of native soil N (p

Labile carbon (C) input to soil can accelerate or slow the decomposition of soil organic matter, a phenomenon called priming. However, priming is difficult to predict, making its relationship with C input elusive. To assess this relationship, we added 13C-glucose at five levels (8 to 1606 μg C g−1 week−1) to the soil from four different ecosystems for seven weeks. We observed a positive linear relationship between C input and priming in all soils: priming increased from negative or no priming at low C input to strong positive priming at high C input. However, the sensitivity of priming to C input varied among soils and between ways of expressing C input, and decreased with elevation. Positive substrate thresholds were detected in three soils (56 to 242 μg C g−1 week−1), suggesting the minimum C input required to trigger positive priming. Carbon input expressed as a fraction of microbial biomass explained 16.5% less variation in priming than did C input expressed as a fraction of dry soil mass, indicating that priming is not strongly related to the size of the soil microbial biomass. We conclude that priming increases with the rate of labile C input, once that rate exceeds a threshold, but the magnitude of increase varies among soils. Further research on mechanisms causing the variation of priming sensitivity to increasing labile C input might help promote a quantitative understanding of how such phenomenon impacts soil C cycling, offering the potential to improve earth system models.

Most soil bacterial taxa are thought to be dormant, or inactive, yet the extent to which they synthetize new rRNA is poorly understood. We analyzed 18O composition of RNA extracted from soil incubated with H218O and used quantitative stable isotope probing to characterize rRNA synthesis among microbial taxa. RNA was not fully labeled with 18O, peaking at a mean of 23.6 ± 6.8 atom percent excess (APE) 18O after eight days of incubation, suggesting some ribonucleotides in soil were more than eight days old. Microbial taxa varied in the degree they incorporated 18O into their rRNA over time and there was no correlation between the APE 18O of bacterial rRNA and their rRNA to DNA ratios, suggesting that the ratios were not appropriate to measure ribonucleotide synthesis. Our study indicates that, on average, 94% of soil taxa produced new rRNA and therefore were metabolically active.

Growth of soil microorganisms is often described as carbon limited, and adding labile carbon to soil often results in a transient and large increase in respiration. In contrast, soil microbial biomass changes little, suggesting that growth and respiration are decoupled in response to a carbon pulse. Alternatively, measuring bulk responses of the entire community (total respiration and biomass) could mask ecologically important variation among taxa in response to the added carbon. Here, we assessed taxon-specific variation in cellular growth (measured as DNA synthesis) and metabolic activity (measured as rRNA synthesis) following glucose addition to soil using quantitative stable isotope probing with H218 O. We found that glucose addition altered rates of DNA and rRNA synthesis, but the effects were strongly taxon specific: glucose stimulated growth and rRNA transcription for some taxa, and suppressed these for others. These contrasting taxon-specific responses could explain the small and transient changes in total soil microbial biomass. Responses to glucose were not well predicted by a priori assignments of taxa into copiotrophic or oligotrophic categories. Across all taxa, rates of DNA and rRNA synthesis changed in parallel, indicating that growth and activity were coupled, and the degree of coupling was unaffected by glucose addition. This pattern argues against the idea that labile carbon addition causes a large reduction in metabolic growth efficiency; rather, the large pulse of respiration observed with labile substrate addition is more likely to be the result of rapid turnover of microbial biomass, possibly due to trophic interactions. Our results support a strong connection between rRNA synthesis and bacterial growth, and indicate that taxon-specific responses among soil bacteria can buffer responses at the scale of the whole community.

Elevated CO2 (eCO(2)) experiments provide critical information to quantify the effects of rising CO2 on vegetation 1-6 . Many eCO(2) experiments suggest that nutrient limitations modulate the local magnitude of the eCO(2) effect on plant biomass(1,3,5), but the global extent of these limitations has not been empirically quantified, complicating projections of the capacity of plants to take up CO27,9. Here, we present a data-driven global quantification of the eCO(2) effect on biomass based on 138 eCO(2) experiments. The strength of CO2 fertilization is primarily driven by nitrogen (N) in similar to 65% of global vegetation and by phosphorus (P) in similar to 25% of global vegetation, with N- or P-limitation modulated by mycorrhizal association. Our approach suggests that CO2 levels expected by 2100 can potentially enhance plant biomass by 12 +/- 3% above current values, equivalent to 59 +/- 13 PgC. The globalscale response to eCO(2) we derive from experiments is similar to past changes in greenness(9) and bio-mass(10) with rising CO2, suggesting that CO2 will continue to stimulate plant biomass in the future despite the constraining effect of soil nutrients. Our research reconciles conflicting evidence on CO2 fertilization across scales and provides an empirical estimate of the biomass sensitivity to eCO(2) that may help to constrain climate projections.

Higher temperatures in northern latitudes will increase permafrost thaw and stimulate above- and belowground plant biomass growth in tundra ecosystems. Higher plant productivity increases the input of easily decomposable carbon (C) to soil, which can stimulate microbial activity and increase soil organic matter decomposition rates. This phenomenon, known as the priming effect, is particularly interesting in permafrost because an increase in C supply to deep, previously frozen soil may accelerate decomposition of C stored for hundreds to thousands of years. The sensitivity of old permafrost C to priming is not well known; most incubation studies last less than one year, and so focus on fast-cycling C pools. Furthermore, the age of respired soil C is rarely measured, even though old C may be vulnerable to labile C inputs. We incubated soil from a moist acidic tundra site in Eight Mile Lake, Alaska for 409 days at 15 °C. Soil from surface (0–25cm), transition (45–55cm), and permafrost (65–85cm) layers were amended with three pulses of uniformly ¹³C-labeled glucose or cellulose every 152 days. Glucose addition resulted in positive priming in the permafrost layer 7 days after each substrate addition, eliciting a two-fold increase in cumulative soil C loss relative to unamended soils with consistent effects across all three pulses. In the transition and permafrost layers, glucose addition significantly decreased the age of soil-respired CO₂-C with Δ¹⁴C values that were 115‰ higher. Previous field studies that measured the age of respired C in permafrost regions have attributed younger Δ¹⁴C ecosystem respiration values to higher plant contributions. However, the results from this study suggest that positive priming, due to an increase in fresh C supply to deeply thawed soil layers, can also explain the respiration of younger C observed at the ecosystem scale. We must consider priming effects to fully understand permafrost C dynamics, or we risk underestimating the contribution of soil C to ecosystem respiration.

We describe a protocol for investigating microbial growth in environmental samples via stable isotope probing (SIP) with H218O. Water is a universal substrate for all microorganisms and replication is required for DNA to become labeled with 18O. By measuring how much the DNA of each taxon becomes enriched with 18O when an environmental sample is incubated with H218O, it is feasible to quantify that population's DNA replication rate, which is a proxy for growth.

Quantitative stable isotope probing (qSIP) measures rates of taxon-specific element assimilation in intact microbial communities, utilizing substrates labeled with a heavy isotope.The laboratory protocol for qSIP is nearly identical to that for conventional stable isotope probing, with two key additions: (1) in qSIP, qPCR measurements are conducted on each density fraction recovered after isopycnic separation, and (2) in qSIP, multiple density fractions are sequenced spanning the entire range of densities over which nucleic acids were recovered. qSIP goes beyond identifying taxa assimilating a substrate, as it also allows for measuring that assimilation for each taxon within a given microbial community. Here, we describe an analysis process necessary to determine atom fraction excess of a heavy stable isotope added to an environmental sample for a given taxon's DNA.

International audience; The continuously increasing concentration of atmospheric CO2 has considerably altered ecosystem functioning. However, few studies have examined the long-term (i.e. over a decade) effect of elevated CO2 on soil microbial communities. Using 16S rRNA gene amplicons and a GeoChip microarray, we investigated soil microbial communities from a Californian annual grassland after 14 years of experimentally elevated CO2 (275 ppm higher than ambient). Both taxonomic and functional gene compositions of the soil microbial community were modified by elevated CO2. There was decrease in relative abundance for taxa with higher ribosomal RNA operon (rrn) copy number under elevated CO2, which is a functional trait that responds positively to resource availability in culture. In contrast, taxa with lower rrn copy number were increased by elevated CO2. As a consequence, the abundance-weighted average rrn copy number of significantly changed OTUs declined from 2.27 at ambient CO2 to 2.01 at elevated CO2. The nitrogen (N) fixation gene nifH and the ammonium-oxidizing gene amoA significantly decreased under elevated CO2 by 12.6% and 6.1%, respectively. Concomitantly, nitrifying enzyme activity decreased by 48.3% under elevated CO2, albeit this change was not significant. There was also a substantial but insignificant decrease in available soil N, with both nitrate (NO3) (-27.4%) and ammonium (NH4+) (-15.4%) declining. Further, a large number of microbial genes related to carbon (C) degradation were also affected by elevated CO2, whereas those related to C fixation remained largely unchanged. The overall changes in microbial communities and soil N pools induced by long-term elevated CO2 suggest constrained microbial N decomposition, thereby slowing the potential maximum growth rate of the microbial community.

Carbon (C) buried deep in soil (below 1 m) is often hundreds to thousands of years old, though the stability and sensitivity of this deep C to environmental change are not well understood. We examined the C dynamics in three soil horizons and their responses to changes in substrate availability in a coarse-textured sandy spodosol (0.0–0.1, 1.0–1.3, and 2.7–3.0 m deep). Substrate additions were intended to mimic an increase in root exudates and available inorganic nitrogen (N) that would follow an increase of belowground biomass at depth, as previously found in a long-term CO2 enrichment experiment at this site. We incubated these soils for 60 days with glucose, alanine, and leaf litter, crossed with an inorganic N amendment equivalent to three times ambient concentrations. The organic substrates were isotopically labeled (13C), allowing us to determine the source of mineralized C and assess the priming effect. Enzyme activity increased as much as 13 times in the two deeper horizons (1.0–1.3, and 2.7–3.0 m) after the addition of the organic substrates, even though the deepest horizon had microbial biomass and microbial phospholipid fatty acids below the level of detection before the experiment. The deepest horizon (2.7–3.0 m) yielded the largest priming response under alanine, indicating that microorganisms in these soil horizons can become active in response to input of organic substrates. Inorganic N amendments significantly decreased the priming effect, suggesting that decomposition may not be N limited. However, alanine (organic N) yielded the highest priming effect at every soil depth, indicating the importance of differentiating effect of organic and inorganic N on decomposition. Distinct priming effects with depth suggest that portions of the soil profile can respond differently to organic inputs. Our findings indicate that the deep soil C pools might be more vulnerable to environmental or anthropogenic change than previously thought, potentially influencing net CO2 exchange estimates between the land and the atmosphere.

Organic matter input to soils can accelerate the decomposition of native soil carbon (C), a process called the priming effect. Priming is ubiquitous and exhibits some consistent patterns, but a general explanation remains elusive, in part because of variation in the response across different ecosystems, and because of a diversity of proposed mechanisms, including microbial activation, stoichiometry, and community shifts. Here, we conducted five-week incubations of four soils (grassland, pinon-juniper, ponderosa pine, mixed conifer), varying the amount of substrate added (as 13C-glucose, either 350 or 1000 μg C g−1 week−1) and either with no added nitrogen (N), or with sufficient N (as NH4NO3) to bring the C-to-N ratio of the added substrate to 10. Using four different ecosystems enabled testing the generality of mechanisms underlying the priming effect. The responses of priming to the amount and C-to-N ratio of the added substrate were consistent across ecosystems: priming increased with the rate of substrate addition and declined when the C-to-N ratio of the substrate was reduced. However, structural equation models failed to confirm intermediate responses postulated to mediate the priming effect, including responses postulated to be mediated by stoichiometry and microbial activation. Specifically, priming was not clearly associated with changes in microbial biomass or turnover, nor with extracellular enzyme activities or the microbial C-to-N ratio. The strongest explanatory pathways in the structural equation models were the substrate, soil, and C-to-N ratio treatments themselves, with no intermediates, suggesting that either these measurements lacked sufficient sensitivity to reveal causal relationships, or the actual drivers for priming were not included in the ancillary measurements. While we observed consistent changes in priming caused by the amount and C-to-N ratio of the added substrate across a wide array of soils, our findings did not clearly conform to common models offered for the priming effect. Because priming is a residual flux involving diverse substrates of varying chemical composition, a simple and generalizable explanation of the phenomenon may be elusive.

Polychlorinated biphenyls (PCBs) are a group of synthetic, lipophilic organochlorines that were banned due to their impacts on human and wildlife health and environmental persistence. Although banned, the continued release from pre-banned products allows them to persist at toxic levels in the environment. This is especially the case in lipid rich food webs of the Arctic, where PCBs accumulate due to both long-range atmospheric transport and locally contaminated sites such as formerly used defense (FUD) sites. At the request of the leadership of the Qawalangin Tribe of Unalaska Island in the Aleutian Archipelago, we analyzed PCB concentrations in samples of threespine stickleback (Gasterosteus aculeatus) and subsistence foods (i.e., salmonid species and blue mussels [Mytilus edulis]) collected at both FUD and non-FUD sites. PCBs were extracted from samples using a QuEChERS method. The mean PCB concentrations across all mussel samples was 6.1 ppb; mussels from FUD sites had nearly double the PCB concentrations (7.6 ppb) compared to non-military sites (3.9 ppb), and at two FUD sites the PCB concentrations exceeded safe consumption guidelines. The mean total PCB concentration for fish was 2.8 ppb; fish PCB concentrations were higher at FUD sites (3.2 ppb) compared to non-military sites (1.2 ppb). These results support the need to remediate the FUD sites of “Building 551/T Dock to Airport” and “Delta Western”. More generally, these results provide further evidence of the continued problem of PCB contamination at FUD sites in the Arctic, many of which are co-located with indigenous communities., Seafood; Food toxicology; Environmental science; Environmental chemistry; Environmental pollution; Water pollution; Environmental toxicology; Public health; Formerly used defense site, FUD, Persistent organic pollutants, POPs, Arctic, Subsistence, Indigenous communities, Fish, Mussels

Nitrogen-induced suppression of lignin-modifying enzyme activity contributes to soil carbon sequestration., Agricultural and industrial activities have increased atmospheric nitrogen (N) deposition to ecosystems worldwide. N deposition can stimulate plant growth and soil carbon (C) input, enhancing soil C storage. Changes in microbial decomposition could also influence soil C storage, yet this influence has been difficult to discern, partly because of the variable effects of added N on the microbial enzymes involved. We show, using meta-analysis, that added N reduced the activity of lignin-modifying enzymes (LMEs), and that this N-induced enzyme suppression was associated with increases in soil C. In contrast, N-induced changes in cellulase activity were unrelated to changes in soil C. Moreover, the effects of added soil N on LME activity accounted for more of the variation in responses of soil C than a wide range of other environmental and experimental factors. Our results suggest that, through responses of a single enzyme system to added N, soil microorganisms drive long-term changes in soil C accumulation. Incorporating this microbial influence on ecosystem biogeochemistry into Earth system models could improve predictions of ecosystem C dynamics.

Growing bacteria have a high concentration of ribosomes to ensure sufficient protein synthesis, which is necessary for genome replication and cellular division. To elucidate whether metabolic activity of soil microorganisms is coupled with growth, we investigated the relationship between rRNA and DNA synthesis in a soil bacterial community using quantitative stable isotope probing (qSIP) with H 2 18 O. Most soil bacterial taxa were metabolically active and grew, and there was no significant difference between the isotopic composition of DNA and RNA extracted from soil incubated with H 2 18 O. The positive correlation between 18 O content of DNA and rRNA of taxa, with a slope statistically indistinguishable from 1 (slope = 0.96; 95% confidence interval [CI], 0.90 to 1.02), indicated that few taxa made new rRNA without synthesizing new DNA. There was no correlation between rRNA-to-DNA ratios obtained from sequencing libraries and the atom percent excess (APE) 18 O values of DNA or rRNA, suggesting that the ratio of rRNA to DNA is a poor indicator of microbial growth or rRNA synthesis. Our results support the notion that metabolic activity is strongly coupled to cellular division and suggest that nondividing taxa do not dominate soil metabolic activity. IMPORTANCE Using quantitative stable isotope probing of microbial RNA and DNA with H 2 18 O, we show that most soil taxa are metabolically active and grow because their nucleic acids are significantly labeled with 18 O. A majority of the populations that make new rRNA also grow, which argues against the common paradigm that most soil taxa are dormant. Additionally, our results indicate that relative sequence abundance-based RNA-to-DNA ratios, which are frequently used for identifying active microbial populations in the environment, underestimate the number of metabolically active taxa within soil microbial communities.

The efficiency with which microbes use substrate (Carbon Use Efficiency or CUE) to make new microbial biomass is an important variable in soil and ecosystem C cycling models. It is generally assumed that CUE of microbial activity in soils is low, however measured values vary widely. It is hypothesized that high values of CUE observed in especially short-term incubations reflect the build-up of storage compounds in response to a sudden increase in substrate availability and are therefore not representative of CUE of microbial activity in unamended soil. To test this hypothesis, we measured the 13CO2 release from six position-specific 13C-labeled glucose isotopomers in ponderosa pine and pinon-juniper soil. We compared this position-specific CO2 production pattern with patterns expected for 1) balanced microbial growth (synthesis of all compounds needed to build new microbial cells) at a low, medium, or high CUE, and 2) synthesis of storage compounds (glycogen, tri-palmitoyl-glycerol, and polyhydroxybutyrate). Results of this study show that synthesis of storage compounds is not responsible for the observed high CUE. Instead, it is the position-specific CO2 production expected for balanced growth and high CUE that best matches the observed CO2 production pattern in these two soils. Comparison with published studies suggests that the amount of glucose added in this study is too low and the duration of the experiment too short to affect microbial metabolism. We conclude that the hypothesis of high CUE in undisturbed soil microbial communities remains viable and worthy of further testing.

Molybdenum (Mo) is critical for the function of enzymes related to nitrogen cycling. Concentrations of Mo are very low in sandy, acidic soils, and biologically available Mo is only a small fraction of the total pool. While several methods have been proposed to measure plant-available Mo, there has not been a recent comprehensive analytical study that compares soil extraction methods as predictors of plant Mo uptake. A suite of five assays [total acid microwave digestion, ethylenediamenetetraaacetic acid (EDTA) extraction, Environmental Protection Agency (EPA) protocol 3050B, ammonium oxalate extraction, and pressurized hot water] was employed, followed by the determination of soil Mo concentrations via inductively coupled mass spectroscopy. The concentrations of soil Mo determined from these assays and their relationships as predictors of plant Mo concentration were compared. The assays yielded different concentrations of Mo: total digest > EPA > ammonium oxalate ≥ EDTA > pressurized hot water. Legume fol...

This is the author accepted manuscript. The final version is available from American Chemical Society via the DOI in this record.

Crowther et al. reported that the best predictor of surface soil carbon (top 10 cm) losses in response to warming is the size of the surface carbon stock in the soil (that is, carbon stocks in plots that have not been warmed), finding that soils that are high in soil carbon also lose more carbon under warming conditions. This relationship was based on a linear regression of soil carbon losses and soil carbon stocks in field warming studies, which was then used to project carbon losses over time and to generate a map of soil carbon vulnerability. However, a few extreme data points (high-leverage points) can strongly influence the slope of a regression line. Only 5 of the 49 sites analysed by Crowther et al.1 are in the upper half of the carbon stock range, which raises the possibility that the relationship they observed could be substantially altered by introducing data from sites with relatively high surface soil carbon stocks. There is a Reply to this Comment by Crowther, T. W. et al. Nature 554, 10.1038/nature25746 (2018).

This is the author accepted manuscript. The final version is available from American Chemical Society via the DOI in this record.

Rising atmospheric CO2 concentrations may alter the nitrogen (N) content of ecosystems by changing N inputs and N losses, but responses vary in field experiments, possibly because multiple mechanisms are at play. We measured N fixation and N losses in a subtropical oak woodland exposed to 11 years of elevated atmospheric CO2 concentrations. We also explored the role of herbivory, carbon limitation, and competition for light or nutrients in shaping the response of N fixation to elevated CO2. Elevated CO2 did not significantly alter gaseous N losses, but lower recovery and deeper distribution in the soil of a long-term 15N tracer indicated that elevated CO2 increased leaching losses. Elevated CO2 had no effect on nonsymbiotic N fixation, and had a transient effect on symbiotic N fixation by the dominant legume. Elevated CO2 tended to reduce soil and plant concentrations of iron, molybdenum, phosphorus, and vanadium, nutrients essential for N fixation. Competition for nutrients and herbivory likely contributed to the declining response of N fixation to elevated CO2. These results indicate that positive responses of N fixation to elevated CO2 may be transient and that chronic exposure to elevated CO2 can increase N leaching. Models that assume increased fixation or reduced N losses with elevated CO2 may overestimate future N accumulation in the biosphere.

Leaf litter provides an important nutrient subsidy to headwater streams, but little is known about how tree genetics influence energy pathways from litter to higher trophic levels. Despite the charge to quantify carbon (C) and nitrogen (N) pathways from decomposing litter, the relationship between litter decomposition and aquatic consumers remains unresolved. We measured litter preference (attachments to litter), C and N assimilation rates, and growth rates of a shredding caddisfly (Hesperophylax magnus, Limnephilidae) in response to leaf litter of different chemical and physical phenotypes using Populus cross types (P. fremontii, P. angustifolia, and F1 hybrids) and genotypes within P. angustifolia. We combined laboratory mesocosm studies using litter from a common garden with a field study using doubly labeled litter (13 C and 15 N) grown in a greenhouse and incubated in Oak Creek, Arizona, USA. We found that, in the lab, shredders initially chose relatively labile (low lignin and condensed tannin concentrations, rapidly decomposing) cross type litter, but preference changed within 4 d to relatively recalcitrant (high lignin and condensed tannin concentrations, slowly decomposing) litter types. Additionally, in the lab, shredder growth rates were higher on relatively recalcitrant compared to labile cross type litter. Over the course of a three-week field experiment, shredders also assimilated more C and N from relatively recalcitrant compared to labile cross type litter. Finally, among P. angustifolia genotypes, N assimilation by shredders was positively related to litter lignin and C:N, but negatively related to condensed tannins and decomposition rate. C assimilation was likewise positively related to litter C:N, and also to litter %N. C assimilation was not associated with condensed tannins or lignin. Collectively, these findings suggest that relatively recalcitrant litter of Populus cross types provides more nutritional benefit, in terms of N fluxes and growth, than labile litter, but among P. angustifolia genotypes the specific trait of litter recalcitrance (lignin or tannins) determines effects on C or N assimilation. As shredders provide nutrients and energy to higher trophic levels, the influence of these genetically based plant decomposition pathways on shredder preference and performance may affect community and food web structure.

Chinese milk vetch (Astragalus sinicus L., vetch), a leguminous winter cover crop, has been widely adopted by farmers in southern China to boost yield of the succeeding rice crop. However, the effects of vetch on rice grain yield and nitrogen (N) use efficiency have not yet been well studied in the intensive double-cropped rice cropping systems. To fill this gap, we conducted a three-year field experiment to evaluate the impacts of the vetch crop on yields and N use efficiency in the subsequent early and late rice seasons. With moderate N input (100 kg N ha&minus, 1 for each rice crop), vetch cover significantly increased grain yields by 7.3&ndash, 13.4% for early rice, by 8.2&ndash, 10.4% for late rice, and by 8.6&ndash, 11.5% for total annual rice production when compared with winter fallow. When rice crops received an N input of 200 kg N ha&minus, 1, vetch cover increased grain yields by 5.9&ndash, 18.4% for early rice, by 3.8&ndash, 10.1% for late rice, and by 6.2&ndash, 11.3% for annual rice production. Moreover, comparable grain yields (11.9 vs. 12.0 Mg ha&minus, 1 for annual rice production) were observed between vetch cover with moderate N and fallow with added N fertilizer. Yield components analysis indicated that the increased tillering number was the main factor for the enhanced grain yields by vetch cover. Vetch cover with moderate and higher N input resulted in higher agronomic N use efficiency and applied N recovery efficiency compared with the fallow treatments. Here, our results showed that vetch as a winter cover crop can be combined with reduced N fertilizer input while maintaining high grain yields, thus gaining a more sustainable rice production system.

Summary Rising atmospheric carbon dioxide (CO2) could alter the carbon (C) and nitrogen (N) content of ecosystems, yet the magnitude of these effects are not well known. We examined C and N budgets of a subtropical woodland after 11 yr of exposure to elevated CO2. We used open-top chambers to manipulate CO2 during regrowth after fire, and measured C, N and tracer 15N in ecosystem components throughout the experiment. Elevated CO2 increased plant C and tended to increase plant N but did not significantly increase whole-system C or N. Elevated CO2 increased soil microbial activity and labile soil C, but more slowly cycling soil C pools tended to decline. Recovery of a long-term 15N tracer indicated that CO2 exposure increased N losses and altered N distribution, with no effect on N inputs. Increased plant C accrual was accompanied by higher soil microbial activity and increased C losses from soil, yielding no statistically detectable effect of elevated CO2 on net ecosystem C uptake. These findings challenge the treatment of terrestrial ecosystems responses to elevated CO2 in current biogeochemical models, where the effect of elevated CO2 on ecosystem C balance is described as enhanced photosynthesis and plant growth with decomposition as a first-order response.

Invasive species alter ecosystems, threaten native and endangered species, and have negative economic impacts. Knowing where invading individuals are from and when they arrive to a new site can guide management. Here, we evaluated how well the stable hydrogen isotope composition (δ2H) records the recent origin and time since arrival of specimens of the invasive Japanese beetle (Popillia japonica Newman) captured near the Portland International Airport (Oregon, U.S.A.). The δ2H of Japanese beetle specimens collected from sites across the contiguous U.S.A. reflected the δ2H of local precipitation, a relationship similar to that documented for other organisms, and one confirming the utility of δ2H as a geographic fingerprint. Within weeks after experimental relocation to a new isotopic environment, the δ2H of beetles changed linearly with time, demonstrating the potential for δ2H to also mark the timing of arrival to a new location. We used a hierarchical Bayesian model to estimate the recent geographical origin and timing of arrival of each specimen based on its δ2H value. The geographic resolution was broad, with values consistent with multiple regions of origin in the eastern U.S.A., slightly favoring the southeastern U.S.A. as the more likely source. Beetles trapped from 2007-2010 had arrived 30 or more days prior to trapping, whereas the median time since arrival declined to 3-7 days for beetles trapped from 2012-2014. This reduction in the time between arrival and trapping at the Portland International Airport supports the efficacy of trapping and spraying to prevent establishment. More generally, our analysis shows how stable isotopes can serve as sentinels of biological invasions, verifying the efficacy of control measures, or, alternatively, indicating when those measures show signs of failure.

Rice cultivation is one of the largest anthropogenic sources of the greenhouse gas methane. Now a meta-analysis shows that increased atmospheric CO2 (550–743 ppmV) and climate warming (+0.8 °C to +6 °C) can be expected to significantly increase the yield-scaled greenhouse-gas emissions of rice. Increased atmospheric CO2 and rising temperatures are expected to affect rice yields and greenhouse-gas (GHG) emissions from rice paddies1,2,3,4. This is important, because rice cultivation is one of the largest human-induced sources of the potent GHG methane5 (CH4) and rice is the world’s second-most produced staple crop6. The need for meeting a growing global food demand7 argues for assessing GHG emissions from croplands on the basis of yield rather than land area8,9,10, such that efforts to reduce GHG emissions take into consideration the consequences for food production. However, it is unclear whether or how the GHG intensity (that is, yield-scaled GHG emissions) of cropping systems will be affected by future atmospheric conditions. Here we show, using meta-analysis, that increased atmospheric CO2 (ranging from 550 to 743 ppmV) and warming (ranging from +0.8 °C to +6 °C) both increase the GHG intensity of rice cultivation. Increased atmospheric CO2 increased GHG intensity by 31.4%, because CH4 emissions are stimulated more than rice yields. Warming increased GHG intensity by 11.8% per 1 °C, largely owing to a decrease in yield. This analysis suggests that rising CO2 and warming will approximately double the GHG intensity of rice production by the end of the twenty-first century, stressing the need for management practices that optimize rice production while reducing its GHG intensity as the climate continues to change.

Effects of anthropogenic nitrogen (N) deposition and the ability of terrestrial ecosystems to store carbon (C) depend in part on the amount of N retained in the system and its partitioning among plant and soil pools. We conducted a meta-analysis of studies at 48 sites across four continents that used enriched 15N isotope tracers in order to synthesize information about total ecosystem N retention (i.e., total ecosystem 15N recovery in plant and soil pools) across natural systems and N partitioning among ecosystem pools. The greatest recoveries of ecosystem 15N tracer occurred in shrublands (mean, 89.5%) and wetlands (84.8%) followed by forests (74.9%) and grasslands (51.8%). In the short term (< 1 week after 15N tracer application), total ecosystem 15N recovery was negatively correlated with fine-root and soil 15N natural abundance, and organic soil C and N concentration but was positively correlated with mean annual temperature and mineral soil C:N. In the longer term (3-18 months after 15N tracer application), total ecosystem 15N retention was negatively correlated with foliar natural-abundance 15N but was positively correlated with mineral soil C and N concentration and C:N, showing that plant and soil natural-abundance 15N and soil C:N are good indicators of total ecosystem N retention. Foliar N concentration was not significantly related to ecosystem 15N tracer recovery, suggesting that plant N status is not a good predictor of total ecosystem N retention. Because the largest ecosystem sinks for 15N tracer were below ground in forests, shrublands, and grasslands, we conclude that growth enhancement and potential for increased C storage in aboveground biomass from atmospheric N deposition is likely to be modest in these ecosystems. Total ecosystem 15N recovery decreased with N fertilization, with an apparent threshold fertilization rate of 46 kg N x ha(-1) x yr(-1) above which most ecosystems showed net losses of applied 15N tracer in response to N fertilizer addition.

Elevated CO2 is expected to lower plant nutrient concentrations via carbohydrate dilution and increased nutrient use efficiency. Elevated CO2 con- sistently lowers plant foliar nitrogen, but there is no consensus on CO2 effects across the range of plant nutrients. We used meta-analysis to quantify elevated CO2 effects on leaf, stem, root, and seed concentrations of B, Ca, Cu, Fe, K, Mg, Mn, P, S, and Zn among four plant functional groups and two levels of N fertiliza- tion. CO2 effects on plant nutrient concentration depended on the nutrient, plant group, tissue, and N status. CO2 reduced B, Cu, Fe, and Mg, but increased Mn concentration in the leaves of N2 fixers. Elevated CO2 increased Cu, Fe, and Zn, but lowered Mn concentration in grass leaves. Tree leaf responses were strongly related to N status: CO2 significantly decreased Cu, Fe, Mg, and S at high N, but only Fe at low N. Elevated CO2 decreased Mg and Zn in crop leaves grown with high N, and Mn at low N. Nutrient concentrations in crop roots were not affected by CO2 enrichment, but CO2 decreased Ca, K, Mg and P in tree roots. Crop seeds had lower S under elevated CO2 .W e also tested the validity of a ''dilution model.'' CO2 reduced the concentration of plant nutrients 6.6% across nutrients and plant groups, but the reduction is less than expected (18.4%) from carbohydrate accu- mulation alone. We found that elevated CO2 impacts plant nutrient status differently among the nutrient elements, plant functional groups, and among plant tissues. Our synthesis suggests that differences between plant groups and plant organs, N status, and differences in nutrient chemistry in soils preclude a universal hypothesis strictly related to carbohydrate dilution regarding plant nutrient response to elevated CO2.

We used metabolic tracers and modeling to analyze the response of soil metabolism to a sudden change in temperature from 4 to 20 °C. We hypothesized that intact soil microbial communities would exhibit shifts in pentose phosphate pathway and glycolysis activity in the same way as is regularly observed for individual microorganisms in pure culture. We also hypothesized that increased maintenance respiration at higher temperature would result in greater energy production and reduced carbon use efficiency (CUE). Two hours after temperature increase, respiration increased almost 10-fold. Although all metabolic processes were increased, the relative activity of metabolic processes, biosynthesis, and energy production changed. Pentose phosphate pathway was reduced (17–20%), while activities of specific steps in glycolysis (51%) and Krebs cycle (7–13%) were increased. In contrast, only small but significant changes in biosynthesis (+2%), ATP production (−3%) and CUE (+2%) were observed. In a second experiment, we compared the metabolic responses to temperature increases in soils from high and low elevation. The shift in activity from pentose phosphate pathway to glycolysis with higher temperature was confirmed in both soils, but the responses of Krebs cycle, biosynthesis, ATP production, and CUE were site dependent. Our results indicate that 1) in response to temperature, communities behave biochemically similarly to single species and, 2) our understanding of temperature effects on CUE, energy production and use for maintenance and growth processes is still incomplete.

Ambient nitrous oxide (N(2)O) emissions from Great Boiling Spring (GBS) in the US Great Basin depended on temperature, with the highest flux, 67.8 ± 2.6 μmol N(2)O-N m(-2) day(-1) , occurring in the large source pool at 82 °C. This rate of N(2)O production contrasted with negligible production from nearby soils and was similar to rates from soils and sediments impacted with agricultural fertilizers. To investigate the source of N(2)O, a variety of approaches were used to enrich and isolate heterotrophic micro-organisms, and isolates were screened for nitrate reduction ability. Nitrate-respiring isolates were identified by 16S rRNA gene sequencing as Thermus thermophilus (31 isolates) and T. oshimai (three isolates). All isolates reduced nitrate to N(2)O but not to dinitrogen and were unable to grow with N(2)O as a terminal electron acceptor. Representative T. thermophilus and T. oshimai strains contained genes with 96-98% and 93% DNA identity, respectively, to the nitrate reductase catalytic subunit gene (narG) of T. thermophilus HB8. These data implicate T. thermophilus and T. oshimai in high flux of N(2)O in GBS and raise questions about the genetic basis of the incomplete denitrification pathway in these organisms and on the fate of biogenic N(2)O in geothermal environments.

Most organic carbon (C) in soils eventually turns into CO2 after passing through microbial metabolic pathways, while providing cells with energy and biosynthetic precursors. Therefore, detailed insight into these metabolic processes may help elucidate mechanisms of soil C cycling processes. Here, we describe a modeling approach to quantify the C flux through metabolic pathways by adding 1-13C and 2,3-13C pyruvate and 1-13C and U-13C glucose as metabolic tracers to intact soil microbial communities. The model calculates, assuming steady-state conditions and glucose as the only substrate, the reaction rates through glycolysis, Krebs cycle, pentose phosphate pathway, anaplerotic activity through pyruvate carboxylase, and various biosynthesis reactions. The model assumes a known and constant microbial proportional precursor demand, estimated from literature data. The model is parameterized with experimentally determined ratios of 13CO2 production from pyruvate and glucose isotopologue pairs. Model sensitivity analysis shows that metabolic flux patterns are especially responsive to changes in experimentally determined 13CO2 ratios from pyruvate and glucose. Calculated fluxes are far less sensitive to assumptions concerning microbial chemical and community composition. The calculated metabolic flux pattern for a young volcanic soil indicates significant pentose phosphate pathway activity in excess of pentose precursor demand and significant anaplerotic activity. These C flux patterns can be used to calculate C use efficiency, energy production and consumption for growth and maintenance purposes, substrate consumption, nitrogen demand, oxygen consumption, and microbial C isotope composition. The metabolic labeling and modeling methods may improve our ability to study the biochemistry and ecophysiology of intact and undisturbed soil microbial communities.

thermophilus narG revealed sediment populations of 1.3-1.7 ¥ 10 6 copies g -1 sediment. These data indicate a highly active nitrogen cycle (N-cycle) in these springs and suggest that ammonia oxidation may be a major source of energy fuelling primary production.