Three‐source partitioning of CO2 efflux from maize field soil by 13C natural abundance

A natural‐¹³C‐labeling approach—formerly observed under controlled conditions—was tested in the field to partition total soil CO₂ efflux into root respiration, rhizomicrobial respiration, and soil organic matter (SOM) decomposition. Different results were expected in the field due to different climate, site, and microbial properties in contrast to the laboratory. Within this isotopic method, maize was planted on soil with C₃‐vegetation history and the total CO₂ efflux from soil was subdivided by isotopic mass balance. The C₄‐derived C in soil microbial biomass was also determined. Additionally, in a root‐exclusion approach, root‐ and SOM‐derived CO₂ were determined by the total CO₂ effluxes from maize (Zea mays L.) and bare‐fallow plots. In both approaches, maize‐derived CO₂ contributed 22% to 35% to the total CO₂ efflux during the growth period, which was comparable to other field studies. In our laboratory study, this CO₂ fraction was tripled due to different climate, soil, and sampling conditions. In the natural‐¹³C‐labeling approach, rhizomicrobial respiration was low compared to other studies, which was related to a low amount of C₄‐derived microbial biomass. At the end of the growth period, however, 64% root respiration and 36% rhizomicrobial respiration in relation to total root‐derived CO₂ were calculated when considering high isotopic fractionations between SOM, microbial biomass, and CO₂. This relationship was closer to the 50% : 50% partitioning described in the literature than without fractionation (23% root respiration, 77% rhizomicrobial respiration). Fractionation processes of ¹³C must be taken into account when calculating CO₂ partitioning in soil. Both methods—natural ¹³C labeling and root exclusion—showed the same partitioning results when ¹³C isotopic fractionation during microbial respiration was considered and may therefore be used to separate plant‐ and SOM‐derived CO₂ sources.     

Sources of CO2 efflux from soil and review of partitioning methods

Five main biogenic sources of CO₂ efflux from soils have been distinguished and described according to their turnover rates and the mean residence time of carbon. They are root respiration, rhizomicrobial respiration, decomposition of plant residues, the priming effect induced by root exudation or by addition of plant residues, and basal respiration by microbial decomposition of soil organic matter (SOM). These sources can be grouped in several combinations to summarize CO₂ efflux from the soil including: root-derived CO₂, plant-derived CO₂, SOM-derived CO₂, rhizosphere respiration, heterotrophic microbial respiration (respiration by heterotrophs), and respiration by autotrophs. These distinctions are important because without separation of SOM-derived CO₂ from plant-derived CO₂, measurements of total soil respiration have very limited value for evaluation of the soil as a source or sink of atmospheric CO₂ and for interpreting the sources of CO₂ and the fate of carbon within soils and ecosystems. Additionally, the processes linked to the five sources of CO₂ efflux from soil have various responses to environmental variables and consequently to global warming. This review describes the basic principles and assumptions of the following methods which allow SOM-derived and root-derived CO₂ efflux to be separated under laboratory and field conditions: root exclusion techniques, shading and clipping, tree girdling, regression, component integration, excised roots and insitu root respiration; continuous and pulse labeling, ¹³C natural abundance and FACE, and radiocarbon dating and bomb-¹⁴C. A short sections cover the separation of the respiration of autotrophs and that of heterotrophs, i.e. the separation of actual root respiration from microbial respiration, as well as methods allowing the amount of CO₂ evolved by decomposition of plant residues and by priming effects to be estimated. All these methods have been evaluated according to their inherent disturbance of the ecosystem and C fluxes, and their versatility under various conditions. The shortfalls of existing approaches and the need for further development and standardization of methods are highlighted.     

Effects of root and litter exclusion on soil CO2 efflux and microbial biomass in wet tropical forests

We examined the effects of root and litter exclusion on the rate of soil CO₂ efflux and microbial biomass at a soil depth of 25 cm in a secondary forest (dominated by Tabebuia heterophylla) and a pine (Pinus caribaea) plantation in the Luquillo Experimental Forest in Puerto Rico. The experimental plots were initially established in 1990, when root, forest floor mass and new litterfall were excluded for 7 y since then. Soil respiration was significantly reduced in the litter and root exclusion plots in both the secondary forest and the pine plantation compared with the control. Root exclusion had a greater effect on soil CO₂ efflux than the litter exclusion in the plantation, whereas a reversed pattern was observed in the secondary forest. The reduction of microbial biomass in the root exclusion plot was greater in the secondary forest (59%) than in the plantation (31%), while there was no difference of the reduction in the litter exclusion plots between these forests. Our results suggest that above-ground input and roots (root litter and exudates) differentially affect soil CO₂ efflux under different vegetation types.     

A new technique to measure soil CO2 efflux at constant CO2 concentration

A new principle for measuring soil CO₂ efflux at constant ambient concentration is introduced. The measuring principle relies on the continuous absorption of CO₂ within the system to achieve a constant CO₂ concentration inside the soil chamber at ambient level, thus balancing the amount of CO₂ entering the soil chamber by diffusion from the soil. We report results that show reliable soil CO₂ efflux measurements with the new system. The novel measuring principle does not disturb the natural gradient of CO₂ within the soil, while allowing for continuous capture of the CO₂ released from the soil. It therefore holds great potential for application in simultaneous measurements of soil CO₂ efflux and its δ¹³C, since both variables show sensitivity to a distortion of the soil CO₂ profile commonly found in conventional chamber techniques.     

Effect of heavy metals contamination on root-derived and organic matter-derived CO2 efflux from soil planted with Zea mays

The CO₂ efflux from loamy Haplic Luvisol and heavy metal (HM) uptake by Zea mays L. were studied under increased HM contamination: Cd, Cu, and Ni up to 20, 1000, and 2500 mg kg⁻¹ soil, respectively. Split-root system with contrasting HM concentrations in both soil halves was used to investigate root-mediated HM translocation in uncontaminated soil zones. To separate root-derived and soil organic matter (SOM)-derived CO₂ efflux from soil, ¹⁴CO₂ pulse labeling of 15-, 25-, and 35-days-old plants was applied. The CO₂ evolution from the bare soil was 10.6 μg C–CO₂ d⁻¹ g⁻¹ (32 kg C–CO₂ d⁻¹ ha⁻¹) and was not affected by HM (except 2500 mg Ni kg⁻¹). The average CO₂ efflux from the soil with maize was about two times higher and amounted for about 22.0 μg C–CO₂ d⁻¹ g⁻¹. Portion of assimilates respired in the rhizosphere decreased with plant development from 6.0 to 7.0% of assimilated C for 25-days-old Zea mays to 0.4–2.0% for 45-days-old maize. The effect of the HM on root-derived ¹⁴CO₂ efflux increased with rising HM content in the following order: Cd < Cu < Ni. In Cu and Ni contaminated soils, shoot and root dry matter decreased to 70% and to 50% of the uncontaminated control, respectively. Plants contained much more HM in the roots than in the shoots. A split-root system with contrasting HM concentrations allowed to trace transport of mobile forms of HM by roots from contaminated soil half into the uncontaminated soil half. The portion of mobile HM forms in the soil (1 M NH₄NO₃ extract) increased with contamination and amounted to 9–16%, 2–6% and 1.5–3.5% for Cd, Cu, and Ni, respectively. Corresponding values for the easily available HM (1 M NH₄OAc extract) were 22–52%, 1–20% and 5–8.5%. Heavy metal availability for plants decreased in the following order: Cd > Cu ≥ Ni. No increase of HM availability in the soil was found after maize cultivation.     

Theoretical model of the abiotic component of soil CO2 tracer efflux in C pulse-labeling experiments on plant-soil systems

For measurement of the time lag between photosynthesis and CO₂ efflux from soil, the carbon isotope pulse-labeling technique is considered as the most suitable. However, an interference from the abiotic tracer CO₂ component is identified as a key difficulty for obtaining accurate results with this technique. Guidelines on how to reduce this interference are therefore urgently needed. The flux of abiotic ¹³CO₂ tracer into soil during the labeling stage, and its return to atmosphere during the monitoring stage was modeled numerically, and the labeling stage also analytically. The controls of the abiotic interference were investigated using these models. The amount of the abiotic tracer component and the time distribution of its rate of return to the atmosphere, were predicted by these models. The main model parameters were D_m (=the ratio between the soil ¹³CO₂ diffusivity and the retardation factor), and the ¹³CO₂ concentration at the soil-atmosphere interface during the labeling stage (S₁₃), while background ¹³CO₂ soil production parameters were unnecessary. The presented models guide the selection of experimental parameters for minimization of the abiotic interference. With parameterization for a particular case, the present numerical model provides a preliminary order-of-magnitude estimate of the abiotic component, which would indicate if this interference is of significance.     

C signature of CO2 evolved from incubated maize residues and maize-derived sheep faeces

Analyses of the spatial and temporal variations in the natural abundance of ¹³C are frequently employed to study transformations of plant residues and soil organic matter turnover on sites where long continued vegetation with the C₃-type photosynthesis pathway has been replaced with a C₄-type vegetation (or vice versa). One controversial issue associated with such analyses is the significance of isotopic fractionation during the microbial turnovers of C in complex substrates. To evaluate this issue, C₃-soil and quartz sand were amended with maize residues and with faeces from sheep feed exclusively on maize silage. The samples were incubated at 15 °C for 117 days (maize residues) or 224 days (sheep faeces). CO₂ evolved during incubation was trapped in NaOH and analysed for C isotopic contents. At the end of incubation, 63 and 50% of the maize C was evolved as CO₂ in the soil and sand, respectively, while 32% of the faeces C incubated with soil and with sand was recovered as CO₂. Maize and faeces showed a similar decomposition pattern but maize decomposed twice as fast as faeces. The δ¹³C of faeces was 0.3‰ lower than that of the maize residue (δ¹³C −13.4‰), while the δ¹³C of the C₃-soil used for incubation was −31.6‰. The δ¹³C value of the CO₂ recovered from unamended C₃-soil was similar or slightly lower (up to −1.5‰) than that of the C₃-soil itself except for an initial flush of ¹³C enriched CO₂. The δ¹³C values of the CO₂ from sand-based incubations typically ranged −15‰ to −17‰, i.e. around −3‰ lower than the δ¹³C measured for maize and faeces. Our study clearly demonstrates that the decomposition of complex substrates is associated with isotopic fractionation, causing evolved CO₂ to be depleted in ¹³C relative to substrates. Consequently the microbial products retained in the soil must be enriched in ¹³C.     

Carbon fluxes in soil food webs of increasing complexity revealed by C labelling and C natural abundance

Soil food webs are mainly based on three primary carbon (C) sources: root exudates, litter, and recalcitrant soil organic matter (SOM). These C sources vary in their availability and accessibility to soil organisms, which could lead to different pathways in soil food webs. The presence of three C isotopes (¹²C, ¹³C and ¹⁴C) offers an unique opportunity to investigate all three C sources simultaneously. In a microcosm experiment we studied the effect of food web complexity on the utilization of the three carbon sources. We choose an incomplete three factorial design with (i) living plants, (ii) litter and (iii) food web complexity. The most complex food web consisted of autochthonous microorganisms, nematodes, collembola, predatory mites, endogeic and anecic earthworms. We traced C from all three sources in soil, in CO₂ efflux and in individual organism groups by using maize grown on soil developed under C₃ vegetation and application of ¹⁴C labelled ryegrass shoots as a litter layer. The presence of living plants had a much greater effect on C pathways than food web complexity. Litter decomposition, measured as ¹⁴CO₂ efflux, was decreased in the presence of living plants from 71% to 33%. However, living plants increased the incorporation of litter C into microbial biomass and arrested carbon in the litter layer and in the upper soil layer. The only significant effect of food web complexity was on the litter C distribution in the soil layers. In treatments with fungivorous microarthropods (Collembola) the incorporation of litter carbon into mineral soil was reduced. Root exudates as C source were passed through rhizosphere microorganisms to the predator level (at least to the third trophic level). We conclude that living plants strongly affected C flows, directly by being a source of additional C, and indirectly by modifying the existing C flows within the food web including CO₂ efflux from the soil and litter decomposition.     

Soil CO2 efflux vs. soil respiration: Implications for flux models

In the long term, all CO₂ produced in the soil must be emitted by the surface and soil CO₂ efflux (FCO₂) must correspond to soil respiration (R_soil). In the short term, however, the efflux can deviate from the instantaneous soil respiration, if the amount of CO₂ stored in the soil pore-space (SCO₂) is changing. We measured FCO₂ continuously for one year using an automated chamber system. Simultaneously, vertical soil profiles of CO₂ concentration, moisture, and temperature were measured in order to assess the changes in the amount of CO₂ stored in the soil. R_soil was calculated as the sum of the rate of change of the CO₂ storage over time and FCO₂. The experiment was split into a warm and a cold season. The dependency of soil respiration and soil efflux on soil temperature and on soil moisture was analyzed separately. Only the moisture-driven model of the warm season was significantly different for FCO₂ and R_soil. At our site, a moisture-driven soil-respiration model derived from CO₂ efflux data would underestimate the importance of soil moisture. This effect can be attributed to a temporary storage of CO₂ in the soil pore-space after rainfalls where up to 40% of the respired CO₂ were stored.     

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.     

Uncoupling of microbial CO2 production and release in frozen soil and its implications for field studies of arctic C cycling

Knowledge of seasonal trends and controls of soil CO₂ emissions to the atmosphere is important for simulating atmospheric CO₂ concentrations and for understanding and predicting the global carbon cycle. This is particularly the case for high arctic soils subject to extreme fluctuating environmental conditions. Based on field measurements of soil CO₂ efflux, temperature, water content, pore gas composition in soil and frozen cores as well as detailed temperature experiments performed in the laboratory, we evaluated seasonal controls of CO₂ effluxes from a well-drained tundra heath site in NE-Greenland. During the growing season, near-surface temperatures correlated well with observed CO₂ effluxes (r²>0.9). However, during intensive thawing of near-surface layers we observed up to 1.5-fold higher effluxes than expected due to temperature alone. These high rates were consistent with high CO₂ concentrations in frozen soil (>10% CO₂) and suggested a spring burst event during soil thawing and a corresponding trapping of produced CO₂ during winter. Laboratory experiments revealed that microbial soil respiration continued down to a least −18 °C and that up to 80% of the produced CO₂ was trapped in soil at temperatures between 0 and −9 °C. The trapping of CO₂ in frozen soil was positively correlated with soil moisture (r²=0.85) and led to an abrupt change of the temperature sensitivity (Q₁₀) observed for soil CO₂ release at 0 °C with Q₁₀ values below 0 °C being up to 100-fold higher than above 0 °C. The results of sub-zero CO₂ production allowed us to predict the microbial soil respiration throughout the year and to evaluate to what extent burst events during thawing can be explained by the release of CO₂ being produced and trapped during winter. Taking only the upper 20 cm of the soil into account, winter soil respiration accounted for about 40% of the annual soil respiration. At least 14% of the winter CO₂ production was trapped during the winter 2000-2001 and observed to be released upon thawing. Thus, the site-specific winter soil respiration is an important part of the annual C cycle and CO₂ trapping should be accounted for in future field and modelling studies of soil respiration dynamics in arctic ecosystems. In conclusion, we have discovered a soil moisture dependent uncoupling of CO₂ production and release in frozen soils with important implications for future field studies of Arctic C cycling.     

Measurement of soil CO2 efflux using soda lime absorption: both quantitative and reliable

Measurement of soil CO₂ efflux using a non-flow-through steady-state (NFT-SS) chamber with alkali absorption of CO₂ by soda lime was tested and compared with a flow-through non-steady-state (FT-NSS) IRGA method to assess suitability of using soda lime for field monitoring over large spatial scales and integrated over a day. Potential errors and artifacts associated with the soda lime chamber method were investigated and improvements made. The following issues relating to quantification and reliable measurement of soil CO₂ efflux were evaluated: (i) absorption capacity of the soda lime, (ii) additional and thus artifactual absorption of CO₂ by soda lime during the experimental procedure, (iii) variation in the CO₂ concentration inside the chamber headspace, and (iv) effects of chamber closure on soil CO₂ efflux. Soil CO₂ efflux, as measured using soda lime (with a range of quantities: 50, 100, and 200 g per 0.082 m² ground area enclosed in chamber), was compared with transient IRGA measurements as a reference method that is based on well-established physical principles, using several forms of spatial and temporal comparisons. Natural variation in efflux rates ranged from 2 to 5.5 g C m⁻² day⁻¹ between different chambers and over different days. A comparison of the IRGA-based assay with measurement based on soda lime yielded an overall correlation coefficient of 0.82. The slope of the regression line was not significantly different from the 1:1 line, and the intercept was not significantly different from the origin. This result indicated that measurement of CO₂ efflux by soda lime absorption was quantitatively similar and unbiased in relation to the reference method. The soda lime method can be a highly practical method for field measurements if implemented with due care (in terms of drying and weighing soda lime, and in minimizing leakages), and validated for specific field conditions. A detailed protocol is presented for use of the soda lime method for measurement of CO₂ efflux from field soils.     

C and N natural abundance of the soil microbial biomass

Stable isotope analysis is a powerful tool in the study of soil organic matter formation. It is often observed that more decomposed soil organic matter is ¹³C, and especially ¹⁵N-enriched relative to fresh litter and recent organic matter. We investigated whether this shift in isotope composition relates to the isotope composition of the microbial biomass, an important source for soil organic matter. We developed a new approach to determine the natural abundance C and N isotope composition of the microbial biomass across a broad range of soil types, vegetation, and climates. We found consistently that the soil microbial biomass was ¹⁵N-enriched relative to the total (3.2 ‰) and extractable N pools (3.7 ‰), and ¹³C-enriched relative to the extractable C pool (2.5 ‰). The microbial biomass was also ¹³C-enriched relative to total C for soils that exhibited a C3-plant signature (1.6 ‰), but ¹³C-depleted for soils with a C4 signature (−1.1 ‰). The latter was probably associated with an increase of annual C3 forbs in C4 grasslands after an extreme drought. These findings are in agreement with the proposed contribution of microbial products to the stabilized soil organic matter and may help explain the shift in isotope composition during soil organic matter formation.     

Natural abundance δN and δC of DNA extracted from soil

We report the first simultaneous measurements of δ¹⁵N and δ¹³C of DNA extracted from surface soils. The isotopic composition of DNA differed significantly among nine different soils. The δ¹³C and δ¹⁵N of DNA was correlated with δ¹³C and δ¹⁵N of soil, respectively, suggesting that the isotopic composition of DNA is strongly influenced by the isotopic composition of soil organic matter. However, in all samples DNA was enriched in ¹³C relative to soil, indicating microorganisms fractionated C during assimilation or preferentially used ¹³C enriched substrates. Enrichment of DNA in ¹⁵N relative to soil was not consistently observed, but there were significant differences between δ¹⁵N of DNA and δ¹⁵N of soil for three different sites, suggesting microorganisms are fractionating N or preferentially using N substrates at different rates across these contrasting ecosystems. There was a strong linear correlation between δ¹⁵N of DNA and δ¹⁵N of the microbial biomass, which indicated DNA was depleted in ¹⁵N relative to the microbial biomass by approximately 3.4‰. Our results show that accurate and precise isotopic measurements of C and N in DNA extracted from the soil are feasible, and that these analyses may provide powerful tools for elucidating C and N cycling processes through soil microorganisms.     

Effects of elevated CO2 concentration on rhizodeposition from Lolium perenne grown on soil exposed to 9 years of CO2 enrichment

The effects of enriched CO₂ atmosphere on partitioning of recently assimilated carbon were investigated in a plant-soil-microorganism system in which Lolium perenne seedlings were planted into cores inserted into the resident soil within a sward that had been treated with elevated CO₂ for 9 consecutive years, under two N fertilisation levels (Swiss FACE experiment). The planted cores were excavated from the ambient (35 Pa pCO₂) and enriched (60 Pa pCO₂) rings at two dates, in spring and autumn, during the growing season. The cores were brought back to the laboratory for ¹⁴C labelling of shoots in order to trace the transfer of recently assimilated C both within the plant and to the soil and microbial biomass. At the spring sampling, high N supply stimulated shoot and total dry matter production. Consistently, high N enhanced the allocation of recently fixed C to shoots, and reduced it to belowground compartments. Elevated CO₂ had no consequences for DM or the pattern of C allocation. At the autumn sampling, at high N plot, yield of L. perenne was stimulated by elevated CO₂. Consistently, ¹⁴C was preferentially allocated aboveground and, consequently belowground recent C allocation was depressed and rhizodeposition reduced. At both experimental periods, total soil C content was similar in all treatments, providing no evidence for soil carbon sequestration in the Swiss Free Air CO₂ Enrichment experiment (FACE) after 9 years of enrichment. Recently assimilated C and soil C were mineralised faster in soils from enriched rings, suggesting a CO₂-induced shift in the microbial biomass characteristics (structure, diversity, activity) and/or in the quality of the root-released organic compounds.     

Carbon dynamics in a temperate grassland soil after 9 years exposure to elevated CO2 (Swiss FACE)

Elevated pCO₂ increases the net primary production, C/N ratio, and C input to the soil and hence provides opportunities to sequester CO₂-C in soils to mitigate anthropogenic CO₂. The Swiss 9 y grassland FACE (free air carbon-dioxide enrichment) experiment enabled us to explore the potential of elevated pCO₂ (60 Pa), plant species (Lolium perenne L. and Trifolium repens L.) and nitrogen fertilization (140 and 540 kg ha⁻¹ y⁻¹) on carbon sequestration and mineralization by a temperate grassland soil. Use of ¹³C in combination with respired CO₂ enabled the identification of the origins of active fractions of soil organic carbon. Elevated pCO₂ had no significant effect on total soil carbon, and total soil carbon was also independent of plant species and nitrogen fertilization. However, new (FACE-derived depleted ¹³C) input of carbon into the soil in the elevated pCO₂ treatments was dependent on nitrogen fertilization and plant species. New carbon input into the top 15 cm of soil from L. perennne high nitrogen (LPH), L. perenne low nitrogen (LPL) and T. repens low nitrogen (TRL) treatments during the 9 y elevated pCO₂ experiment was 9.3±2.0, 12.1±1.8 and 6.8±2.7 Mg C ha⁻¹, respectively. Fractions of FACE-derived carbon in less protected soil particles >53 μm in size were higher than in <53 μm particles. In addition, elevated pCO₂ increased CO₂ emission over the 118 d incubation by 55, 61 and 13% from undisturbed soil from LPH, LPL and TRL treatments, respectively; but only by 13, 36, and 18%, respectively, from disturbed soil (without roots). Higher input of new carbon led to increased decomposition of older soil organic matter (priming effect), which was driven by the quantity (mainly roots) of newly input carbon (L. perenne) as well as the quality of old soil carbon (e.g. higher recalcitrance in T. repens). Based on these results, the potential of well managed and established temperate grassland soils to sequester carbon under continued increasing concentrations of atmospheric CO₂ appears to be rather limited.     

Elevated CO2 concentration and nitrogen fertilisation effects on N2O and CH4 fluxes and biomass production of Phleum pratense on farmed peat soil

The effects of elevated CO₂ supply on N₂O and CH₄ fluxes and biomass production of Phleum pratense were studied in a greenhouse experiment. Three sets of 12 farmed peat soil mesocosms (10 cm dia, 47 cm long) sown with P. pratense and equally distributed in four thermo-controlled greenhouses were fertilised with a commercial fertiliser in order to add 2, 6 or 10 g N m⁻². In two of the greenhouses, CO₂ concentration was kept at atmospheric concentration (360 μmol mol⁻¹) and in the other two at doubled concentration (720 μmol mol⁻¹). Soil temperature was kept at 15 °C and air temperature at 20 °C. Natural lighting was supported by artificial light and deionized water was used to regulate soil moisture. Forage was harvested and the plants fertilised three times during the basic experiment, followed by an extra fertilisations and harvests. At the end of the experiment CH₄ production and CH₄ oxidation potentials were determined; roots were collected and the biomass was determined. From the three first harvests the amount of total N in the aboveground biomass was determined. N₂O and CH₄ exchange was monitored using a closed chamber technique and a gas chromatograph. The highest N₂O fluxes (on average, 255 μg N₂O m⁻² h⁻¹ during period IV) occurred just after fertilisation at high water contents, and especially at the beginning of the growing season (on average, 490 μg N₂O m⁻² h⁻¹ during period I) when the competition of vegetation for N was low. CH₄ fluxes were negligible throughout the experiment, and for all treatments the production and oxidation potentials of CH₄ were inconsequential. Especially at the highest rates of fertilisation, the elevated supply of CO₂ increased above- and below-ground biomass production, but both at the highest and lowest rates of fertilisation, decreased the total amount of N in the aboveground dry biomass. N₂O fluxes tended to be higher under doubled CO₂ concentrations, indicating that increasing atmospheric CO₂ concentration may affect N and C dynamics in farmed peat soil.     

Availability of CO2 as a factor affecting the rate of nitrification in soil

A laboratory incubation experiment was conducted to demonstrate that reduced availability of CO₂ in soil may be an important factor limiting nitrification. Soil samples were incubated at 30±2 °C for 20 days using vessels with or without the arrangement for trapping CO₂ in sodium hydroxide. This arrangement led to a decrease of ca. 96% in the CO₂ concentration of the headspace, with a range of 95.7-97.5 at different sampling intervals. In the absence of trapping arrangement, CO₂ concentration of the headspace varied from 580 to 859 ppm, i.e. 62-140% higher than that of the outside atmosphere (358 ppm). The nitrification process was significantly retarded under conditions of reduced CO₂ concentration; reduction varied from 8 to 62% at different incubation intervals. The results of the study led to the inference that decreased availability of CO₂ in closed vessels (with arrangement for trapping CO₂) will have a significant bearing on the process of nitrification and hence on the overall dynamics of N transformations.     

Impact of atmospheric CO2 and plant life forms on soil microbial activities

From the global change perspective, increase of atmospheric CO₂ and land cover transformation are among the major impacts caused by human activities. In this study, we are addressing the combined issues of the effect of CO₂ concentration increase and plant type on soil microbial activities by asking how annual and perennial plant groups affect soil microbial processes under elevated CO₂. The experimental design used a mix of species of different growth forms for both annuals and perennials. Our objective was: (1) to determine how two years of annual or perennial plant cover and CO₂ enrichment could affect Mediterranean soil microbial processes; (2) to test the resistance and the resilience of these soil functional processes after a natural perturbation. We determined the effects of 2 years atmospheric CO₂ enrichment on soil potential respiration (SIR), denitrification (DEA) and nitrification (NEA) activities. We could not find any significant effect of CO₂ increase on SIR, DEA and NEA. However, we found a strong effect of the plant cover type, i.e. annuals versus perennials, on the potential microbial activity related to N cycling. DEA and NEA were significantly higher in soil under annual plants while SIR was not significantly different. To determine whether these changes would survive a natural perturbation, we carried out a rain event experiment once the experimental treatments (i.e. different plant cover and atmospheric CO₂ concentration) were stopped. The soil potential respiration, as expressed by the SIR, was not affected and remained stable. DEA rates converged rapidly under annuals and perennials after the rain event. Under both annuals and perennials NEA increased significantly after the rain event but remained significantly higher in the soil with annual plants. The relative change of the soil microbial processes induced by annual and perennial plants was inversely related to the density and the diversity of the corresponding microbial functional groups.     

Winter soil CO2 efflux and its contribution to annual soil respiration in different ecosystems of a forest-steppe ecotone, north China

Most soil respiration measurements are conducted during the growing season. In tundra and boreal forest ecosystems, cumulative winter soil CO₂ fluxes are reported to be a significant component of their annual carbon budgets. However, little information on winter soil CO₂ efflux is known from mid-latitude ecosystems. Therefore, comparing measurements of soil respiration taken annually versus during the growing season will improve the accuracy of ecosystem carbon budgets and the response of soil CO₂ efflux to climate changes. In this study we measured winter soil CO₂ efflux and its contribution to annual soil respiration for seven ecosystems (three forests: Pinus sylvestris var. mongolica plantation, Larix principis-rupprechtii plantation and Betula platyphylla forest; two shrubs: Rosa bella and Malus baccata; and two meadow grasslands) in a forest-steppe ecotone, north China. Overall mean winter and growing season soil CO₂ effluxes were 0.15-0.26 μmol m⁻² s⁻¹ and 2.65-4.61 μmol m⁻² s⁻¹, respectively, with significant differences in the growing season among the different ecosystems. Annual Q₁₀ (increased soil respiration rate per 10 °C increase in temperature) was generally higher than the growing season Q₁₀. Soil water content accounted for 84% of the variations in growing season Q₁₀ and soil temperature range explained 88% of the variation in annual Q₁₀. Soil organic carbon density to 30 cm depth was a good surrogate for SR₁₀ (basal soil respiration at a reference temperature of 10 °C). Annual soil CO₂ efflux ranged from 394.76 g C m⁻² to 973.18 g C m⁻² using observed ecosystem-specific response equations between soil respiration and soil temperature. Estimates ranged from 424.90 g C m⁻² to 784.73 g C m⁻² by interpolating measured soil respiration between sampling dates for every day of the year and then computing the sum to obtain the annual value. The contributions of winter soil CO₂ efflux to annual soil respiration were 3.48-7.30% and 4.92-7.83% using interpolated and modeled methods, respectively. Our results indicate that in mid-latitude ecosystems, soil CO₂ efflux continues throughout the winter and winter soil respiration is an important component of annual CO₂ efflux.