Soil organic matter dynamics in grassland soils under elevated CO2: Insights from long-term incubations and stable isotopes

Elevated atmospheric carbon dioxide (CO₂) levels generally stimulate carbon (C) uptake by plants, but the fate of this additional C largely remains unknown. This uncertainty is due in part to the difficulty in detecting small changes in soil carbon pools. We conducted a series of long-term (170-330 days) laboratory incubation experiments to examine changes in soil organic matter pool sizes and turnover rates in soil collected from an open-top chamber (OTC) elevated CO₂ study in Colorado shortgrass steppe. We measured concentration and isotopic composition of respired CO₂ and applied a two-pool exponential decay model to estimate pool sizes and turnover rates of active and slow C pools. The active and slow C pools of surface soils (5-10 cm depth) were increased by elevated CO₂, but turnover rates of these pools were not consistently altered. These findings indicate a potential for C accumulation in near-surface soil C pools under elevated CO₂. Stable isotopes provided evidence that elevated CO₂ did not alter the decomposition rate of new C inputs. Temporal variations in measured δ¹³C of respired CO₂ during incubation probably resulted mainly from the decomposition of changing mixtures of fresh residue and older organic matter. Lignin decomposition may have contributed to declining δ¹³C values late in the experiments. Isotopic dynamics during decomposition should be taken into account when interpreting δ¹³C measurements of soil respiration. Our study provides new understanding of soil C dynamics under elevated CO₂ through the use of stable C isotope measurements during microbial organic matter mineralization.     

Effects of nitrogen fertilization on pot‐grown wheat photosynthate partitioning within intensively farmed soil determined by 13C pulse‐labeling

Understanding rhizodeposited carbon (C) dynamics of winter wheat (Triticum aestivum L.) is important for improving soil fertility and increasing soil C stocks. However, the effects of nitrogen (N) fertilization on photosynthate C allocation to rhizodeposition of wheat grown in an intensively farmed alkaline soil remain elusive. In this study, pot‐grown winter wheat under N fertilization of 250 kg N ha^?1 was pulse‐labeled with ¹³CO₂ at tillering, elongation, anthesis, and grain‐filling stages. The ¹³C in shoots, roots, soil organic carbon (SOC), and rhizosphere‐respired CO₂ was measured 28 d after each ¹³C labeling. The proportion of net‐photosynthesized ¹³C recovered (shoots + roots + soil + soil respired CO₂) in the shoots increased from 58–64% at the tillering to 86–91% at the grain‐filling stage. Likewise, the proportion in the roots decreased from 21–28% to 2–3%, and that in the SOC pool increased from 1–2% to 6–7%. However, the ¹³C respired CO₂ allocated to soil peaked (17–18%) at the elongation stage and decreased to 6–8% at the grain‐filling stage. Over the entire growth season of wheat, N fertilization decreased the proportion of net photosynthate C translocated to the below‐ground pool by about 20%, but increased the total amount of fixed photosynthate C, and therefore increased the below‐ground photosynthate C input. We found that the chase period of about 4 weeks is sufficient to accurately monitor the recovery of ¹³C after pulse labeling in a wheat–soil system. We conclude that N fertilization increased the deposition of photoassimilate C into SOC pools over the entire growth season of wheat compared to the control treatment.     

Turnover of soil organic matter and of microbial biomass under C3-C4 vegetation change: Consideration of C fractionation and preferential substrate utilization

Two processes contribute to changes of the δ¹³C signature in soil pools: ¹³C fractionation per se and preferential microbial utilization of various substrates with different δ¹³C signature. These two processes were disentangled by simultaneously tracking δ¹³C in three pools - soil organic matter (SOM), microbial biomass, dissolved organic carbon (DOC) - and in CO₂ efflux during incubation of 1) soil after C₃-C₄ vegetation change, and 2) the reference C₃ soil.The study was done on the Ap horizon of a loamy Gleyic Cambisol developed under C₃ vegetation. Miscanthus giganteus - a perennial C₄ plant - was grown for 12 years, and the δ¹³C signature was used to distinguish between ‘old’ SOM (>12 years) and ‘recent’ Miscanthus-derived C (<12 years). The differences in δ¹³C signature of the three C pools and of CO₂ in the reference C₃ soil were less than 1‰, and only δ¹³C of microbial biomass was significantly different compared to other pools. Nontheless, the neglecting of isotopic fractionation can cause up to 10% of errors in calculations. In contrast to the reference soil, the δ¹³C of all pools in the soil after C₃-C₄ vegetation change was significantly different. Old C contributed only 20% to the microbial biomass but 60% to CO₂. This indicates that most of the old C was decomposed by microorganisms catabolically, without being utilized for growth. Based on δ¹³C changes in DOC, CO₂ and microbial biomass during 54 days of incubation in Miscanthus and reference soils, we concluded that the main process contributing to changes of the δ¹³C signature in soil pools was preferential utilization of recent versus old C (causing an up to 9.1‰ shift in δ¹³C values) and not ¹³C fractionation per se.Based on the δ¹³C changes in SOM, we showed that the estimated turnover time of old SOM increased by two years per year in 9 years after the vegetation change. The relative increase in the turnover rate of recent microbial C was 3 times faster than that of old C indicating preferential utilization of available recent C versus the old C.Combining long-term field observations with soil incubation reveals that the turnover time of C in microbial biomass was 200 times faster than in total SOM. Our study clearly showed that estimating the residence time of easily degradable microbial compounds and biomarkers should be done at time scales reflecting microbial turnover times (days) and not those of bulk SOM turnover (years and decades). This is necessary because the absence of C reutilization is a prerequisite for correct estimation of SOM turnover. We conclude that comparing the δ¹³C signature of linked pools helps calculate the relative turnover of old and recent pools.     

Three-source-partitioning of microbial biomass and of CO2 efflux from soil to evaluate mechanisms of priming effects

We propose and successfully applied a new approach for 3-source-partitioning based on a combination of ¹⁴C labeling with ¹³C natural abundance. By adding ¹⁴C-labeled glucose to soil after C₃ - C₄ vegetation change, we partitioned three C sources in three compartments, namely CO₂, microbial biomass and dissolved organic C (DOC). This enabled us to estimate mechanisms and sources of priming effects (PE).Glucose application at low and high rate (GL: 100 and GH: 1000 μg C g⁻¹, respectively) caused positive PE both short-term (during 1-3 days) and long-term (3-55 days). Despite a 10-fold difference in the amount of substrate added, the PE observed was larger by a factor of only 1.6 at the high versus low rate of glucose. The real and apparent priming effects were distinguished by partitioning of microbial C for glucose-C and SOM-derived C. As the amount of primed CO₂ respired during short-term PE was 40% lower than microbial C, and the contribution of soil C in microbial biomass did not increase, we concluded that such short-term PE was apparent and was mainly caused by accelerated microbial turnover (at GL) and by pool substitution (at GH). Both the amount of primed CO₂-C, which was 1.3-2.1 times larger than microbial C, and the increased contribution of soil C in microbial biomass allowed us to consider the long-term PE as being real. The sole source of real PE (GL treatment) was the “recent” soil organic matter, which is younger than 12-year-old C. The real PE-induced by a glucose amount exceeding microbial biomass (GH) was due to the almost equal contribution of ‘recent’ (<12 years) and ‘old’ (>12 years) C. Thus, the decomposition of old recalcitrant SOM was induced only by an amount of primer exceeding microbial C. We conclude that combining ¹⁴C labeling with ¹³C natural abundance helped disentangle three C sources in CO₂, microbial biomass and DOC and evaluate mechanisms and sources of PE.     

Soil carbon dioxide flux, carbon sequestration and crop productivity in a tropical dryland agroecosystem: Influence of organic inputs of varying resource quality

In view of the significance of agricultural soils in affecting global C balance, the impact of manipulation of the quality of exogenous inputs on soil CO₂–C flux was studied in rice–barley annual rotation tropical dryland agroecosystem. Chemical fertilizer, Sesbania shoot (high quality resources), wheat straw (low quality resource) and Sesbania + wheat straw (high + low quality), all carrying equivalent recommended dose of N, were added to soil. A distinct seasonal variation in CO₂–C flux was recorded in all treatments, flux being higher during rice period, and much reduced during barley and summer fallow periods. During rice period the mean CO₂–C flux was greater in wheat straw (161% increase over control) and Sesbania + wheat straw (+129%) treatments; however, during barley and summer fallow periods differences among treatments were small. CO₂–C flux was more influenced by seasonal variations in water-filled pore space compared to soil temperature. In contrast, the role of microbial biomass and live crop roots in regulating soil CO₂–C flux was highly limited. Wheat straw input showed smaller microbial biomass with a tendency of rapid turnover rate resulting in highest cumulative CO₂–C flux. The Sesbania input exhibited larger microbial biomass with slower turnover rate, leading to lower cumulative CO₂–C flux. Addition of Sesbania to wheat straw showed higher cumulative CO₂–C flux yet supported highest microbial biomass with lowest turnover rate indicating stabilization of microbial biomass. Although single application of wheat straw or Sesbania showed comparable net change in soil C (18% and 15% relative to control, respectively) and crop productivity (32% and 38%), yet they differed significantly in soil C balance (374 and −3 g C m⁻² y⁻¹ respectively), a response influenced by the recalcitrant and labile nature of the inputs. Combining the two inputs resulted in significant increment in net change in soil C (33% over control) and crop yield (49%) in addition to high C balance (152 g C m⁻² y⁻¹). It is suggested that appropriate mixing of high and low quality inputs may contribute to improved crop productivity and soil fertility in terms of soil C sequestration.     

Microbial response to the addition of glucose in low-fertility soils

Addition of soluble organic substrates to soil has been shown to either increase or restrict the rate of microbial CO₂–C evolution. This has been attributed to a priming effect resulting from accelerated or decreased turnover of the soil organic matter including the soil microflora. We investigated microbial responses to small glucose-C additions (10–50 μg C g¹ soil) in arable soils either amended or not with cellulose. An immediate CO₂–C release between 0 and 69 h (equivalent to 59% of glucose-C applied) was measured. However, only half of the CO₂–C respired could be attributed to the utilisation of glucose-C substrate, based on the percentage of ¹⁴C–CO₂ evolved after the addition of a ¹⁴C-labelled glucose tracer. Thus, although no evidence of an immediate release of ‘extra’ C above the rate applied as glucose-C was observed, the pattern of decomposition for ¹⁴C-glucose suggested utilisation of an alternate C source. Based on this, a positive priming effect (1.5 to 4.3 times the amount CO₂–C evolved that was attributed to glucose-C decomposition) was observed for at least 170 h in non-cellulose-amended soil and 612 h in cellulose-amended soil. Two further phases of microbial activity in cellulose-amended soils were attributed to either activation of different microbial populations or end-product inhibition of cellulase activity after glucose addition. During these subsequent phases, a negative priming effect of between 0.1 and 1.5 times was observed. Findings indicate that the response of the microbial community to small additions of soluble organic C substrate is not consistent and support the premise that microbial response varies in a yet to be predicted manner between soil type and ecosystems. We hypothesise that this is due to differences in the microbial community structure activated by the addition of organic C and the timing of soluble organic substrate addition with respect to the current dissolved organic C status of the soil.     

Using metabolic tracer techniques to assess the impact of tillage and straw management on microbial carbon use efficiency in soil

Tillage practices and straw management can affect soil microbial activities with consequences for soil organic carbon (C) dynamics. Microorganisms metabolize soil organic C and in doing so gain energy and building blocks for biosynthesis, and release CO₂ to the atmosphere. Insight into the response of microbial metabolic processes and C use efficiency (CUE; microbial C produced per substrate C utilized) to management practices may therefore help to predict long term changes in soil C stocks. In this study, we assessed the effects of reduced (RT) and conventional tillage (CT) on the microbial central C metabolic network, using soil samples from a 12-year-old field experiment in an Irish winter wheat cropping system. Straw was removed from half of the RT and CT plots after harvest or incorporated into the soil in the other half, resulting in four treatment combinations. We added 1-¹³C and 2,3-¹³C pyruvate and 1-¹³C and U-¹³C glucose as metabolic tracer isotopomers to composite soil samples taken at two depths (0–15 cm and 15–30 cm) from each of the treatments and used the rate of position-specific respired ¹³CO₂ to parameterize a metabolic model. Model outcomes were then used to calculate CUE of the microbial community. Whereas the composite samples differed in CUE, the changes were small, with values ranging between 0.757 and 0.783 across treatments and soil depth. Increases in CUE were associated with a reduced tricarboxylic acid cycle and reductive pentose phosphate pathway activity and increased consumption of metabolic intermediates for biosynthesis. Our results suggest that RT and straw incorporation do not substantially affect CUE.     

Influence of soil phosphorus status and nitrogen addition on carbon mineralization from 14C-labelled glucose in pasture soils

 This study examines the effect of soil P status and N addition on the decomposition of ¹⁴C-labelled glucose to assess the consequences of reduced fertilizer inputs on the functioning of pastoral systems. A contrast in soil P fertility was obtained by selecting two hill pasture soils with different fertilizer history. At the two selected sites, representing low (LF) and high (HF) fertility status, total P concentrations were 640 and 820 mg kg^–1 and annual pasture production was 4,868 and 14,120 kg DM ha^–1 respectively. Soils were amended with ¹⁴C-labelled glucose (2,076 mg C kg^–1 soil), with and without the addition of N (207 mg kg^–1 soil), and incubated for 168 days. During incubation, the amounts of ¹⁴CO₂ respired, microbial biomass C and ¹⁴C, microbial biomass P, extractable inorganic P (P_i) and net N mineralization were determined periodically. Carbon turnover was greatly influenced by nutrient P availability. The amount of glucose-derived ¹⁴CO₂ production was high (72%) in the HF and low (67%) in the LF soil, as were microbial biomass C and P concentrations. The ¹⁴C that remained in the microbial biomass at the end of the 6-month incubation was higher in the LF soil (15%) than in the HF soil (11%). Fluctuations in P_i in the LF soil during incubation were small compared with those in HF soil, suggesting that P was cycling through microbial biomass. The concentrations of P_i were significantly greater in the HF samples throughout the incubation than in the LF samples. Net N mineralization and nitrification rates were also low in the LF soils, indicating a slow turnover of microorganisms under limited nutrient supply. Addition of N had little effect on biomass ¹⁴C and glucose utilization. This suggests that, at limiting P fertility, C turnover is retarded because microbial biomass becomes less efficient in the utilization of substrates. Received: 18 October 1999     

Decomposition of C-labeled roots in a pasture soil exposed to 10 years of elevated CO2

The net flux of soil C is determined by the balance between soil C input and microbial decomposition, both of which might be altered under prolonged elevated atmospheric CO₂. In this study, we determined the effect of elevated CO₂ on decomposition of grass root material (Lolium perenne L.). ¹⁴C-labeled root material, produced under ambient (35 Pa pCO₂) or elevated CO₂ (70 Pa pCO₂) was incubated in soil for 64 days. The soils were taken from a pasture ecosystem which had been exposed to ambient (35 Pa pCO₂) or elevated CO₂ (60 Pa pCO₂) under FACE-conditions for 10 years and two fertilizer N rates: 140 and 560 kg N ha⁻¹ year⁻¹. In soil exposed to elevated CO₂, decomposition rates of root material grown at either ambient or elevated CO₂ were always lower than in the control soil exposed to ambient CO₂, demonstrating a change in microbial activity. In the soil that received the high rate of N fertilizer, decomposition of root material grown at elevated CO₂ decreased by approximately 17% after incubation for 64 days compared to root material grown at ambient CO₂. The amount of ¹⁴CO₂ respired per amount of ¹⁴C incorporated in the microbial biomass (q¹⁴CO₂) was significantly lower when roots were grown under high CO₂ compared to roots grown under low CO₂. We hypothesize that this decrease is the result of a shift in the microbial community, causing an increase in metabolic efficiency. Soils exposed to elevated CO₂ tended to respire more native SOC, both with and without the addition of the root material, probably resulting from a higher C supply to the soil during the 10 years of treatment with elevated CO₂. The results show the importance of using soils adapted to elevated CO₂ in studies of decomposition of roots grown under elevated CO₂. Our results further suggest that negative priming effects may obscure CO₂ data in incubation experiments with unlabeled substrates. From the results obtained, we conclude that a slower turnover of root material grown in an ‘elevated-CO₂ world’ may result in a limited net increase in C storage in ryegrass swards.     

Rhizosphere priming effect: Its functional relationships with microbial turnover, evapotranspiration, and C–N budgets

Understanding soil organic matter (SOM) decomposition and its interaction with rhizosphere processes is a crucial topic in soil biology and ecology. Using a natural ¹³C tracer method to separately measure SOM-derived CO₂ from root-derived CO₂, this study aims to connect the level of rhizosphere-dependent SOM decomposition with the C and N balance of the whole plant–soil system, and to mechanistically link the rhizosphere priming effect to soil microbial turnover and evapotranspiration. Results indicated that the magnitude of the rhizosphere priming effect on SOM decomposition varied widely, from zero to more than 380% of the unplanted control, and was largely influenced by plant species and phenology. Balancing the extra soil C loss from the strong rhizosphere priming effect in the planted treatments with C inputs from rhizodeposits and root biomass, the whole plant–soil system remained with a net carbon gain at the end of the experiment. The increased soil microbial biomass turnover rate and the enhanced evapotranspiration rate in the planted treatments had clear positive relationships with the level of the rhizosphere priming effect. The rhizosphere enhancement of soil carbon mineralization in the planted treatments did not result in a proportional increase in net N mineralization, suggesting a possible de-coupling of C cycling with N cycling in the rhizosphere.     

Isotopic analysis of respired CO2 during decomposition of separated soil organic matter pools

A detailed understanding of the processes that contribute to the δ¹³C value of respired CO₂ is necessary to make links between the isotopic signature of CO₂ efflux from the soil surface and various sources within the soil profile. We used density fractionation to divide soils from two forested sites that are a part of an ongoing detrital manipulation experiment (the Detrital Input and Removal Treatments, or DIRT project) into two soil organic matter pools, each of which contributes differently to total soil CO₂ efflux. In both sites, distinct biological pools resulted from density fractionation; however, our results do not always support the concept that the light fraction is readily decomposable whereas the heavy fraction is recalcitrant. In a laboratory incubation following density fractionation we found that cumulative respiration over the course of the incubation period was greater from the light fraction than from the heavy fraction for the deciduous site, while the opposite was true for the coniferous site.Use of stable isotopes yielded insight as to the nature of the density fractions, with the heavy fraction solids from both forests isotopically enriched relative to those of the light fraction. The isotopic signature of respired CO₂, however, was more complicated. During incubation of the fractions there was an initial isotopic depletion of the respired CO₂ compared to the substrate for both soil fractions from both forests. Over time for both fractions of both soils the respired δ¹³C reflected more closely the initial substrate value; however, the transition from depleted to enriched respiration relative to substrate occurs at a different stage of decomposition depending on site and substrate recalcitrance. We found a relationship between cumulative respiration during the incubation period and the duration of the transition from isotopically depleted to enriched respiration in the coniferous site but not the deciduous site. Our results suggest that a shift in microbial community or to dead microbial biomass as a substrate could be responsible for the transition in the isotopic signature of respired CO₂ during decomposition. It is likely that a combination of organic matter quality and isotopic discrimination by microbes, in addition to differences in microbial community composition, contribute to the isotopic signature of different organic matter fractions. It is apparent that respired δ¹³CO₂ cannot be assumed to be a direct representation of the substrate δ¹³C. Detailed knowledge of the soil characteristics at a particular site is necessary to interpret relationships between the isotopic values of a substrate and respired CO₂.     

C fractionation at the root-microorganisms-soil interface: A review and outlook for partitioning studies

Natural variations of the ¹³C/¹²C ratio have been frequently used over the last three decades to trace C sources and fluxes between plants, microorganisms, and soil. Many of these studies have used the natural-¹³C-labelling approach, i.e. natural δ¹³C variation after C₃-C₄ vegetation changes. In this review, we focus on ¹³C fractionation in main processes at the interface between roots, microorganisms, and soil: root respiration, microbial respiration, formation of dissolved organic carbon, as well as microbial uptake and utilization of soil organic matter (SOM). Based on literature data and our own studies, we estimated that, on average, the roots of C₃ and C₄ plants are ¹³C enriched compared to shoots by +1.2 ± 0.6‰ and +0.3 ± 0.4‰, respectively. The CO₂ released by root respiration was ¹³C depleted by about −2.1 ± 2.2‰ for C₃ plants and −1.3 ± 2.4‰ for C₄ plants compared to root tissue. However, only a very few studies investigated ¹³C fractionation by root respiration. This urgently calls for further research. In soils developed under C₃ vegetation, the microbial biomass was ¹³C enriched by +1.2 ± 2.6‰ and microbial CO₂ was also ¹³C enriched by +0.7 ± 2.8‰ compared to SOM. This discrimination pattern suggests preferential utilization of ¹³C-enriched substances by microorganisms, but a respiration of lighter compounds from this fraction. The δ¹³C signature of the microbial pool is composed of metabolically active and dormant microorganisms; the respired CO₂, however, derives mainly from active organisms. This discrepancy and the preferential substrate utilization explain the δ¹³C differences between microorganisms and CO₂ by an ‘apparent’ ¹³C discrimination. Preferential consumption of easily decomposable substrates and less negative δ¹³C values were common for substances with low C/N ratios. Preferential substrate utilization was more important for C₃ soils because, in C₄ soils, microbial respiration strictly followed kinetics, i.e. microorganisms incorporated heavier C (? = +1.1‰) and respired lighter C (? = −1.1‰) than SOM. Temperature and precipitation had no significant effect on the ¹³C fractionation in these processes in C₃ soils. Increasing temperature and decreasing precipitation led, however, to increasing δ¹³C of soil C pools.Based on these ¹³C fractionations we developed a number of consequences for C partitioning studies using ¹³C natural abundance. In the framework of standard isotope mixing models, we calculated CO₂ partitioning using the natural-¹³C-labelling approach at a vegetation change from C₃ to C₄ plants assuming a root-derived fraction between 0% and 100% to total soil CO₂. Disregarding any ¹³C fractionation processes, the calculated results deviated by up to 10% from the assumed fractions. Accounting for ¹³C fractionation in the standard deviations of the C₄ source and the mixing pool did not improve the exactness of the partitioning results; rather, it doubled the standard errors of the CO₂ pools. Including ¹³C fractionations directly into the mass balance equations reproduced the assumed CO₂ partitioning exactly. At the end, we therefore give recommendations on how to consider ¹³C fractionations in research on carbon flows between plants, microorganisms, and soil.     

The initial rate of C substrate utilization and longer-term soil C storage

The initial reaction of microbial transformation and turnover of soil carbon inputs may influence the magnitude of longer-term net soil C storage. The objective of this study was to test the merit of the hypothesis that the more rapid substrates are initially utilized, the longer the residual products remain in the soil. We used simple model C compounds to determine their decomposition rates and persistence over time. Pure ¹⁴C compounds of glucose, acetate, arginine, oxalate, phenylalanine, and urea were incubated in soil for 125 days at 24°C. Total respired CO₂ and ¹⁴CO₂ was quantitatively measured every day for 15 days and residual soil ¹⁴C after 125 days. The percent ¹⁴C remaining in the soil after 125 days of incubation was positively and significantly correlated with the percent substrate utilized in the first day of incubation. The ¹⁴C in the microbial biomass ranged from 4–15% after 15 days and declined through day 125, contributing significantly to the ¹⁴C that evolved over the longer time period. Priming of ¹²C soil organic matter (SOM) was negative at day 3 but became positive, reaching a maximum on day 12; the total increase in soil C from added substrates was greater than the primed C. The primed C came from ¹²C SOM rather than the microbial biomass. This data supports the concept that the more rapidly a substrate is initially mineralized, the more persistent it will be in the soil over time.     

Microbial community utilization of added carbon substrates in response to long-term carbon input manipulation

The chemical composition and quantity of plant inputs to soil are primary factors controlling the size and structure of the soil microbial community. Little is known about how changes in the composition of the soil microbial community affect decomposition rates and other ecosystem functions. This study examined the degradation of universally ¹³C-labeled glucose, glutamate, oxalate, and phenol in soil from an old-growth Douglas-fir (Pseudotsuga menziesii)—western hemlock (Tsuga heterophylla) forest in the Oregon Cascades that has experienced 7 y of chronic C input manipulation. The soils used in this experiment were part of a larger Detritus Input and Removal Treatment experiment and have received normal C inputs (control), doubled wood inputs, or root and litter input exclusion (no inputs). Soil from the doubled wood treatment had a higher fungal:bacterial ratio, and soil from the no inputs treatment had a lower fungal:bacterial ratio, than the control soil. Differences in the utilization of the compounds added to the field-manipulated soils were assessed by following the ¹³C tracer into microbial biomass and respiration. In addition, ¹³C-phospholipid fatty acids (PLFA) analysis was used to examine differential microbial utilization of the added substrates. Glucose and glutamate were metabolized similarly in soils of all three litter treatments. In contrast, the microbial community in the double wood soil respired more added phenol and oxalate, whereas microbes in the no inputs soil respired less added phenol and oxalate, than the control soil. Phenol was incorporated primarily into fungal PLFA, especially in soil of the double wood treatment. The addition of all four substrates led to enhanced degradation of soil organic matter (priming) in soils of all three litter treatments, and was greater following the addition of phenol and oxalate as compared to glucose and glutamate. Priming was greater in the no inputs soil as compared to the control or doubled wood soils. These results demonstrate that altering plant inputs to soil can lead to changes in microbial utilization of C compounds. It appears that many of these changes are the result of alteration in the size and composition of the microbial community.     

Resilience of microbial respiration, respiratory quotient and stable isotope characteristics to soil hydrocarbon addition

On the basis of CO₂ evolution rate, O₂ uptake rate, and ¹³C isotopic signature of respired CO₂, the metabolic response to the addition of ¹³C labelled n-hexadecane and palmitic acid each with supplementary nitrogen was studied for two topsoils, one under continuous agricultural management and the other under beech forest. The CO₂ evolution rate was immediately stimulated in the agricultural soil and the respiratory quotient (RQ) decreased from 0.8 to 0.4 mol CO₂ evolution rate per mol O₂ uptake rate, which was below the theoretically expected value of 0.65 and 0.70 for the degradation of n-hexadecane and palmitic acid, respectively. The microbial response was delayed in the forest soil, but developed better than in the agricultural soil throughout the subsequent 2-4 weeks. Consequently, the respiration rate returned earlier to the initial level for the beech forest soil and the δ¹³C of respired CO₂ and RQ approached values before hydrocarbon addition. Based on the link among respiration rates, RQ and ¹³C-CO₂ value, the added oil-analogue compounds induced a more rapid response in the agricultural soil and were degraded more completely in the forest soil. We concluded that the resilience, which we defined here as the capacity of the soil microbiota to buffer perturbance and to reorganise in response to change resulting in a more desirable system, was higher in our forest soil than for the agricultural soil.     

Drying/rewetting cycles mobilize old C from deep soils from a California annual grassland

We measured the ¹⁴C and ¹³C signatures of CO₂ respired from surface and deep soils released through multiple dry/rewetting cycles in laboratory incubations. The C respired from surface soils included components fixed before and after the 1960s. However, that respired from deep soils was derived from organic matter with a mean turnover time estimated in the range of 650-850 years. This reinforces previous research suggesting that a substantial amount of deep soil C is chemically labile but physically inaccessible to microorganisms, but also suggests that substantial amounts of that C may not be so strongly bound to minerals as to be effectively inert, raising the question of why it hasn’t already been metabolized. It also demonstrates the contribution of C fixed before the 1960s to CO₂ metabolized in the surface soils at this site.     

Is there a linear relationship between priming effect intensity and the amount of organic matter input?

Inputs of fresh organic matter (FOM) are known to affect the rate of soil organic matter (SOM) mineralization. SOM mineralization can be accelerated or decelerated by FOM inputs. This phenomenon, known as the Priming effect (PE), may largely influence the carbon (C) storage capacity of soils. However, the link between PE intensity and FOM inputs is not clearly understood. Indeed, almost all the studies about PE used only one FOM amount which is generally largely below the amount of FOM observed in field conditions. In our study, we incubated soil amended with three levels of ¹³C-labeled straw as FOM and a control without FOM amendment for 80 days. The three levels used were in the same range as the natural FOM inputs observed on our sampling site. Various levels of mineral nitrogen were added within each level of straw supply so that the final input C:N ratios ranged among 44, 30 and 20. CO₂ and δ¹³C-CO₂ were measured during the experiment allowing us to distinguish the FOM respired CO₂ from the SOM respired CO₂. We observed that PE intensity did not increase linearly with increasing FOM additions. Moreover, decreasing the input C:N ratios did not systematically affect PE intensity probably because of shifts in the microbial characteristics such as their C:N ratio or their assimilation yields. These results suggest that PE is a saturating function of FOM inputs that is only weakly influenced by initial N availability. Our results may be explained (i) by the existence of a limited SOM pool subject to PE (ii) or by the occurrence of two simultaneous and antagonistic mechanisms: an increase of the total active microbial biomass accelerating SOM mineralization (i.e. a positive PE) and a preferential substrate utilization of FOM over SOM decreasing SOM mineralization (i.e. a negative PE). Finally, irrespective of the mechanisms implied, our results suggest that the importance of positive PE relatively to the amount of FOM may decrease when FOM inputs increase, which is favorable to carbon sequestration in soils. Indeed, in the case of the lower amount of FOM, the PE corresponded to 6.25% of the total amount of CO₂ mineralized at the end of the experiment while, for the higher amount of FOM, the PE corresponded to 5% of the total amount of CO₂ mineralized at the end of the experiment.     

Input of carbon to soil from wheat plants

The growth of wheat plants and the distribution of labelled photosynthate from pulse-labelling with ¹⁴CO₂ were measured periodically during the growing season in the field. During early growth there was approximately the same proportion of photosynthate translocated below ground and retained in the shoots. Of the ¹⁴C below ground about a half was respired and a quarter each was in the soil and roots. This distribution changed exponentially during growth with an increasing proportion of ¹⁴C remaining in the shoots and a corresponding decreasing proportion being translated below ground, which was only a few percent by flowering. From this information the total input of carbon to the soil from the crop was calculated to be 1305 kg Cha⁻¹. Comparison of these estimates with those from previous experiments suggest that differences do occur due to the stage of growth of the plant, the environmental conditions, soil type and microbial activity.     

A passive sampling method for radiocarbon analysis of soil respiration using molecular sieve

Radiocarbon analysis of soil CO₂ can provide information on the age, source and turnover rate of soil organic C. We developed a new method for passively trapping respired CO₂ on molecular sieve, allowing it to be returned to the laboratory and recovered for C isotope analysis. We tested the method on a soil at a grassland site, and using a synthetic soil created to provide a contrasting isotopic signature. As with other passive sampling techniques, a small amount of fractionation of the ¹³C isotope occurs during sampling, which we have quantified, otherwise the results show that the molecular sieve traps a sufficiently large and representative sample of CO₂ for C isotope analysis. Since ¹⁴C results are routinely corrected for mass-dependent fractionation, our results show that passive sampling of soil respiration using molecular sieve offers a reliable method to collect soil-respired CO₂ for ¹⁴C analysis.     

Identification of groups of metabolically-active rhizosphere microorganisms by stable isotope probing of PLFAs

We combined microbial community phospholipid fatty acid (PLFA) analyses with an in situ stable isotope ¹³CO₂ labelling approach to identify microbial groups actively involved in assimilation of root-derived C in limed grassland soils. We hypothesized that the application of lime would stimulate more rapid ¹³C assimilation and turnover in microbial PLFAs. Four and 8 d after label application, 18:1ω9, 18:2ω6,9 (fungal biomarkers) and 16:1ω7, 18:1ω7, 19:0cy (Gram-negative bacterial biomarkers) showed the most ¹³C enrichment and rapid turnover rates. This suggests that these microorganisms were assimilating recently-photosynthesized root C inputs to soils. Contrary to our hypothesis, liming did not affect assimilation or turnover rates of ¹³C-labelled C. ¹³C stable isotope pulse-labelling technique paired with analyses of PLFA microbial biomarkers shows promise for in situ investigations of microbial function in soils.