Controls over pathways of carbon efflux from soils along climate and black spruce productivity gradients in interior Alaska

Small changes in C cycling in boreal forests can change the sign of their C balance, so it is important to gain an understanding of the factors controlling small exports like water-soluble organic carbon (WSOC) fluxes from the soils in these systems. To examine this, we estimated WSOC fluxes based on measured concentrations along four replicate gradients in upland black spruce (Picea mariana [Mill.] BSP) productivity and soil temperature in interior Alaska and compared them to concurrent rates of soil CO₂ efflux. Concentrations of WSOC in organic and mineral horizons ranged from 4.9 to 22.7 g C m⁻² and from 1.4 to 8.4 g C m⁻², respectively. Annual WSOC fluxes (4.5-12.0 g C m⁻² y⁻¹) increased with annual soil CO₂ effluxes (365-739 g C m⁻² y⁻¹) across all sites (R²=0.55, p=0.02), with higher fluxes occurring in warmer, more productive stands. Although annual WSOC flux was relatively small compared to total soil CO₂ efflux across all sites (<3%), its relative contribution was highest in warmer, more productive stands which harbored less soil organic carbon. The proportions of relatively bioavailable organic fractions (hydrophilic organic matter and low molecular weight acids) were highest in WSOC in colder, low-productivity stands whereas the more degraded products of microbial activity (fulvic acids) were highest in warmer, more productive stands. These data suggest that WSOC mineralization may be a mechanism for increased soil C loss if the climate warms and therefore should be accounted for in order to accurately determine the sensitivity of boreal soil organic C balance to climate change.     

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.     

Use of 13C and 15N natural abundance techniques to characterize carbon and nitrogen dynamics in composting and in compost-amended soils

Isotope fractionation during composting may produce organic materials with a more homogenous δ¹³C and δ¹⁵N signature allowing study of their fate in soil. To verify this, C, N, δ¹³C and δ¹⁵N content were monitored during nine months covered (thermophilic; >40 °C) composting of corn silage (CSC). The C concentration reduced from 10.34 to 1.73 g C (g ash)⁻¹, or 83.3%, during composting. Nitrogen losses comprised 28.4% of initial N content. Compost δ¹³C values became slightly depleted and increasingly uniform (from −12.8±0.6‰ to −14.1±0.0‰) with composting. Compost δ¹⁵N values (0.3±1.3 to 8.2±0.4‰) increased with a similar reduced isotope variability.The fate of C and N of diverse composts in soil was subsequently examined. C, N, δ¹³C, δ¹⁵N content of whole soil (0-5 cm), light (<1.7 g cm⁻³) and heavy (>1.7 g cm⁻³) fraction, and (250-2000 μm; 53-250 μm and <53 μm) size separates, were characterized. Measurements took place one and two years following surface application of CSC, dairy manure compost (DMC), sewage sludge compost (SSLC), and liquid dairy manure (DM) to a temperate (C₃) grassland soil. The δ¹³C values and total C applied (Mg C ha⁻¹) were DM (−27.3‰; 2.9); DMC (−26.6‰; 10.0); SSLC (−25.9‰; 10.9) and CSC (−14.0‰; 4.6 and 9.2). The δ¹³C of un-amended soil exhibited low spatial (−28.0‰±0.2; n=96) and temporal (±0.1‰) variability. All C₄ (CSC) and C₃ (DMC; SSLC) composts, except C₃ manure (DM), significantly modified bulk soil δ¹³C and δ¹⁵N. Estimates of retention of compost C in soil by carbon balance were less sensitive than those calculated by C isotope techniques. One and two years after application, 95 and 89% (CSC), 75 and 63% (SSLC) and 88 and 42% (DMC) of applied compost C remained in the soil, with the majority (80-90%) found in particulate (>53 μm) and light fractions. However, C₄ compost (CSC) was readily detectable (12% of compost C remaining) in mineral (<53 μm) fractions. The δ¹⁵N-enriched N of compost supported interpretation of δ¹³C data. We can conclude that composts are highly recalcitrant with prolonged C storage in non-mineral soil fractions. The sensitivity of the natural abundance tracer technique to characterize their fate in soil improves during composting, as a more homogeneous C isotope signature develops, in addition to the relatively large amounts of stable C applied in composts.     

Organic carbon and stable C isotope in conservation agriculture and conventional systems

Conservation agriculture might have the potential to increase soil organic C content compared to conventional tillage based systems. The present study quantified soil organic carbon (SOC) and soil C derived from C₃ (wheat) and C₄ (maize) plant species using δ¹³C stable isotope. Soil with 16 y of continuous application of zero tillage (ZT) or conventional tillage (CT), monoculture (M) or rotation (R) of wheat and maize, and with (+r) and without retention (−r) in the field of crop residues were studied in the central highlands of Mexico. The highest SOC content was found in the 0-5 cm layer under ZTM and ZTR with residues retention. The soil cultivated with maize showed a higher SOC content in the 0-10 cm layer with residue retention than without residue. In the 10-20 cm layer, the highest SOC content was found in the CT treatment with residue retention. The SOC stock expressed as equivalent soil mass was greatest in the 0-20 cm layer of the ZTM (wheat and maize) and ZTR cultivated treatments with residue retention. After 16 y, the highest content of soil δ¹³C was found in ZTM + r and CTM + r treated soil cultivated with maize; −16.56‰ and −18.08‰ in the 0-5 cm layer, −18.41‰ and −18.02‰ in the 5-10 cm layer and −18.59‰ and −18.72‰ in the 10-20 cm layer respectively. All treatments had a higher percentages of C-C₃ (derived from wheat residues or the earlier forest) than C-C₄ (derived from maize residues). The highest percentages of C-C₄, was found in ZTM + r and CTM + r treated soil cultivated with maize, i.e. 33.0% and 13.0% in 0-5 cm layer, 9.1% and 14.3% in the 5-10 cm layer and 5.0% and 6.8% in 10-20 cm layer, respectively. The gross SOC turnover was lower in soil with residue retention than without residues. It was found that the ZT system with residue retention and rotation with wheat is a practice with a potential to retain organic carbon in soil.     

Use of C abundance to study short-term pig slurry decomposition in the field

Applying pig slurry (PS) on agricultural soils is a common practice. However, its impact on soil organic C dynamics is not clear. This experiment investigated the use of natural ¹³C abundance to study the short-term C mineralization of anaerobically stored PS under field conditions. Measurements of δ¹³C-CO₂ were made on soil air samples obtained from a bare sandy loam during 22 d following incorporation of either PS alone, PS+barley straw, or barley straw alone; an unamended treatment was used as a control. Slurry C was enriched in ¹³C (−20.0‰) because of the high corn (Zea mays L.) content of the animal diet. This value contrasted with δ¹³C of −28.4‰ for the soil organic matter and of −29.0‰ for the barley straw. A peak of high δ¹³CO₂ values (average of −9.2‰) was observed on the day of PS application and was attributed to the dissociation of PS carbonates when mixed with the relatively acidic soil. After this initial burst, 36% of the evolved CO₂ originated from the decomposing PS. After 22 d of incubation, approx. 20% of the PS-C had been lost as CO₂. This short-term field study did not show any priming effect of PS on the mineralization of straw or native soil C. Due to its heterogeneity, the use of the isotopic composition of the evolved CO₂ for estimating PS decomposition requires precaution either through the use of a specific experimental design involving comparable C3 and C4 treatments, or calculations to account for the presence of ¹³C-enriched inorganic C in the PS.     

Spatial heterogeneity in the relocation of added C within the structure of an upland grassland soil

A pulse of ¹³CO₂ was added to the above ground vegetation in an upland grassland to determine the effects of faunal diversity on the flux of carbon to the surface horizons of the soil. Faunal diversity was manipulated by liming and biocide treatments for three years prior to the pulse addition. The relocation of ¹³C within roots and rhizosphere soil was determined by analysis of samples of bulk soil and of specific features identified on soil thin sections on four dates after the addition of the ¹³CO₂ pulse. Analysis of bulk soils showed only a small enrichment in ¹³C and no significant effects of the treatments. Analysis by isotope ratio mass spectrometry of the products of in situ laser combustion of root material and aggregates formed from faunal excrement showed that the distribution of the newly photosynthesised ¹³C is very localised, with large spatial variability in soil and root δ¹³C at scales of less than 1 mm. δ¹³C values ranged from the natural abundance level of around −28‰ to −4.9‰ in roots and to −8.4‰ in aggregates. The small pulse and large spatial variability masked any effects of the liming and biocide treatments in these soils. However, the variability in the relocation of newly photosynthesised carbon may help to explain the large spatial variability found in bacterial numbers at the sub-mm scale within soils and emphasises the importance of the accessibility of substrates to decomposers in undisturbed structured soils.     

Soil C and N pools in patchy shrublands of the Negev and Chihuahuan Deserts

Patchy distribution of vegetation within semi-arid shrublands is normally mirrored in the soil beneath perennial shrubs (macrophytic patches), compared to inter-shrub areas (microphytic patches). To determine impacts of (1) litterfall inputs within vegetation patches and (2) rainfall distribution on soil C and N, we investigated soil C and N pools and associated soil properties in two semi-arid shrublands, in the Negev Desert of Israel (Lehavim), which receives >90% of annual rainfall during winter and in the Chihuahuan Desert, USA (FHMR) that experiences a bimodal (Summer-Winter) annual rainfall pattern. We also evaluated grazing effects on soil C and N pools at Lehavim. More distinct differences in soil properties existed between patch types at the Negev site, where the soils contained higher soil organic C and N, amino acids and sugars, asparaginase activity and plant-available N than those at FHMR. Soil organic C (0-5 cm) in macrophytic patches was 39 g/kg at Lehavim and 13 g/kg at FHMR, and asparaginase activity was as high as 70 μg N/g 2 h in macrophytic patches at Lehavim, two times higher than at FHMR. The soil (0-5 cm) δ¹³C was −15 to −18‰ at Lehavim and −18 to −19‰ at FHMR, with significantly lower δ¹³C in macrophytic patches at both sites. The δ¹³C suggested that considerable macrophytic patch soil C was derived from cyanobacteria at Lehavim and C4 grasses at FHMR. Plant litter δ¹⁵N was 0.9‰ at Lehavim and 0.6‰ at FHMR, suggesting that much plant N was derived from N fixation. Concentrations of inorganic soil N (NH₄⁺+NO₃⁻) were up to 37 mg N/kg at Lehavim and <9 mg N/kg at FHMR. Grazing at Lehavim resulted in lower soil CH, AA, and AS. We conclude that differences between the sites are due largely to (i) higher amounts of litterfall C and N inputs within macrophytic patches at Lehavim and (ii) the different precipitation patterns, with summer precipitation at FHMR promoting increased organic matter mineralization compared to Lehavim, which experiences Winter precipitation only. Furthermore, greater differences in soil properties between patch types at Lehavim compared to FHMR can likely be attributed to the increasing importance of physical processes of resource dispersion at the more humid site in Arizona.     

Rice root-derived carbon input and its effect on decomposition of old soil carbon pool under elevated CO2

Rice (Oryza sativa) was grown in sunlit, semi-closed growth chambers (4×3×2 m, L×W×H) at 650 μl l⁻¹ CO₂ (elevated CO₂) to determine: (1) rice root-derived carbon (C) input into the soil under elevated CO₂ in one growing season, and (2) the effect of the newly input C on decomposition of the more recalcitrant native soil organic C. The initial δ¹³C value of the experimental soil was −25.8‰, which was 6‰ less depleted in ¹³C than the plants grown under elevated CO₂. Significant changes in δ¹³C of the soil organic C were detected after one growing season. The amount of new soil C input was estimated to be 0.9 t ha⁻¹ (or 2.1%) at 30 kg N ha⁻¹ and 1.8 t ha⁻¹ (4.1%) at 90 kg N ha⁻¹. Changes in soil δ¹³C suggested that the surface 5 cm of soil received more C input from plants than soils below. Laboratory incubation (25 °C) of soils from different horizons indicated that increased availability of the labile plant-derived C in the soil reduced decomposition of the native soil organic C. Provided the retardant effect of the new C on old soil organic C holds in the field in the longer-term, paddy soils will likely sequester more C from the atmosphere if more plant C enters the soil under elevated atmospheric CO₂.     

Thermal stability of soil organic matter pools and their δC values after C3-C4 vegetation change

Carbon isotopic composition of soils subjected to C₃-C₄ vegetation change is a suitable tool for the estimation of C turnover in soil organic matter (SOM) pools. We hypothesized that the biological availability of SOM pools is inversely proportional to their thermal stability. Soil samples from a field plot with 10.5 years of cultivation of the C₄ plant Miscanthus×gigantheus and from a reference plot under C₃ grassland vegetation were analysed by thermogravimetry coupled with differential scanning calorimetry (TG-DSC). According to differential weight losses (dTG) and energy release or consumption (DSC), five SOM pools with increasing thermal stability were distinguished: (I) 20-190 °C, (II) 190-310 °C, (III) 310-390 °C, (IV) 390-480 °C, and (V) 480-1000 °C. Their δ¹³C values were analysed by EA-IRMS. The weight losses in pool I were connected with water evaporation, since no significant C losses were measured and δ¹³C values remained unchanged. The δ¹³C of pools II and III in soil samples under Miscanthus were closer to the δ¹³C of the Miscanthus plant tissues (−11.8‰) compared to the thermally stable SOM pool V (−19.5‰). The portion of the Miscanthus-derived C₄-C in total SOM in 0-5 cm reached 55.4% in the 10.5 years. The C₄-C contribution in pool II was 60% and decreased down to 6% in pool V. The mean residence times (MRT) of SOM pools II, III, and IV were similar (11.6, 12.2, and 15.4 years, respectively), while pool V had a MRT of 163 years. Therefore, we concluded that the biological availability of thermal labile SOM pools (<480 °C) was higher, than that of the thermal stable pool decomposed above 480 °C. However, the increase of SOM stability with rising temperature was not gradual. Therefore, the applicability of the TG-DSC for the separation of SOM pools with different biological availability is limited.     

Quantifying the contribution of soil organic matter turnover to forest soil respiration, using natural abundance δC

Quantifying the loss of soil carbon through respiration has proved difficult, due to the challenge of measuring the losses associated with the turnover of soil organic matter (SOM) as distinct from autotrophic components. In forest ecosystems the δ¹³C value of respiration from turnover of SOM (δ¹³CR_SOM) is typically 2-4‰ enriched compared with that from roots and associated microbes (δ¹³CR_ROOTS), with that from the litter (δ¹³CR_LITTER) lying between the two. We measured soil respiration at 50 locations in a forest soil and then used differences in isotopic signatures to quantify the proportion of soil respiration arising from the turnover of SOM (fR_SOM) at a subset of 30 locations, chosen randomly. The soil surface CO₂ efflux was collected using an open chamber system supplied with CO₂-free air and the δ¹³C signature (δ¹³CR_S) measured, giving a mean (±SD) value across the site of −26.1 ± 0.58‰. The values of δ¹³CR_ROOTS, δ¹³CR_LITTER and δ¹³CR_SOM were measured at each location by incubation of roots, litter and root-free soil and collection of the CO₂ for isotopic analysis. δ¹³CR_SOM became progressively depleted with length of incubation (1.5‰ after 8 h), so CO₂ was collected after 20 min. The mean value of δ¹³CR_LITTER was −27.2 ± 0.68 ‰, which was indistinguishable from δ¹³CR_ROOTS of −27.6 ± 0.51‰, while δ¹³CR_SOM was −25.1 ± 0.88‰. δ¹³CR_ROOTS and δ¹³CR_SOM measured at each location were used as the end points of a two component mixing model to calculate fR_SOM, giving a mean value for fR_SOM of 0.61 ± 0.28. It was not possible to estimate fR_SOM using the total C contents of the roots and soil which were significantly depleted in ¹³C in comparison to their respired CO₂. However, at seven locations the δ¹³CR_S was slightly enriched compared with δ¹³CR_SOM (mean 0.3‰), which was not considered significantly different so fR_SOM was constrained to 1.0. If these seven rings were excluded mean fR_SOM was 0.49 ± 0.20. We have shown the possibility of using natural abundance ¹³C discrimination to quantify fR_SOM in a forest soil with an input of carbon only from C₃ photosynthesis.     

Nitrate leaching in soil: Tracing the NO3 sources with the help of stable N and O isotopes

Legumes increase the plant-available N pool in soil, but might also increase NO₃⁻ leaching to groundwater. To minimize NO₃⁻ leaching, N-release processes and the contribution of legumes to NO₃⁻ concentrations in soil must be known. Our objectives were (1) to quantify NO₃⁻-N export to >0.3 m soil depth from three legume monocultures (Medicago x varia Martyn, Onobrychis viciifolia Scop., Lathyrus pratensis L.) and from three bare ground plots. Furthermore, we (2) tested if it is possible to apply a mixing model for NO₃⁻ in soil solution based on its dual isotope signals, and (3) estimated the contribution of legume mineralization to NO₃⁻ concentrations in soil solution under field conditions. We collected rainfall and soil solution at 0.3 m soil depth during 1 year, and determined NO₃⁻ concentrations and δ¹⁵N and δ¹⁸O of NO₃⁻ for >11.5 mg NO₃⁻-N l⁻¹. We incubated soil samples to assess potential N release by mineralization and determined δ¹⁵N and δ¹⁸O signals of NO₃⁻ derived from mineralization of non-leguminous and leguminous organic matter.Mean annual N export to >0.3 m soil depth was highest in bare ground plots (9.7 g NO₃⁻-N m⁻²; the SD reflects the spatial variation) followed by Medicago x varia monoculture (6.0 g NO₃⁻-N m⁻²). The O. viciifolia and L. pratensis monocultures had a much lower mean annual N export (0.5 and 0.3 g NO₃⁻-N m⁻²). The averaged NO₃⁻-N leaching during 70 days was not significantly different between field estimates and incubation for the Medicago x varia Martyn monoculture.The δ¹⁵N and δ¹⁸O values in NO₃⁻ of rainfall (δ¹⁵N: 3.3±0.8‰; δ¹⁸O: 30.8±4.7‰), mineralization of non-leguminous SOM (9.3±0.9‰; 6.7±0.8‰), and mineralization of leguminous SOM (1.5±0.6‰; 5.1±0.9‰) were markedly different. Applying a linear mixing model based on these three sources to δ¹⁵N and δ¹⁸O values in NO₃⁻ of soil solution during winter 2003, we calculated 18-41% to originate from rainfall, 38-57% from mineralization of non-leguminous SOM, and 18-40% from mineralization of leguminous SOM.Our results demonstrate that (1) even under legumes NO₃⁻-N leaching was reduced compared to bare ground, (2) the application of a three-end-member mixing model for NO₃⁻ based on its dual isotope signals produced plausible results and suggests that under particular circumstances such models can be used to estimate the contributions of different NO₃⁻ sources in soil solution, and (3) in the 2nd year after establishment of legumes, they contributed approximately one-fourth to NO₃⁻-N loss.     

Carbon input belowground is the major C flux contributing to leaf litter mass loss: Evidences from a C labelled-leaf litter experiment

Partitioning of the quantities of C lost by leaf litter through decomposition into (i) CO₂ efflux to the atmosphere and (ii) C input to soil organic matter (SOM) is essential in order to develop a deeper understanding of the litter-soil biogeochemical continuum. However, this is a challenging task due to the occurrence of many different processes contributing to litter biomass loss. With the aim of quantifying different fluxes of C lost by leaf litter decomposition, a field experiment was performed at a short rotation coppice poplar plantation in central Italy. Populus nigra leaf litter, enriched in ¹³C (δ¹³C ∼ +160‰) was placed within collars to decompose in direct contact with the soil (δ¹³C ∼ −26‰) for 11 months. CO₂ efflux from within the collars and its isotopic composition were determined at monthly intervals. After 11 months, remaining litter and soil profiles (0-20 cm) were sampled and analysed for their total C and ¹³C content. Gas chromatography (GC), GC-mass spectrometry (MS) and GC-combustion-isotope ratio (GC/C/IRMS) were used to analyse phospholipid fatty acids (PLFA) extracted from soil samples to identify the groups of soil micro-organisms that had incorporated litter-derived C and to determine the quantity of C incorporated by the soil microbial biomass (SMB). By the end of the experiment, the litter had lost about 80% of its original weight. The fraction of litter C lost as an input into the soil (67 ± 12% of the total C loss) was found to be twice as much as the fraction released as CO₂ to the atmosphere (30 ± 3%), thus demonstrating the importance of quantifying litter-derived C input to soils, in litter decomposition studies. The mean δ¹³C values of PLFAs in soil (δ¹³C = −12.5‰) showed sustained incorporation of litter-derived C after one year (7.8 ± 1.6% of total PLFA-C). Thus, through the application of stable ¹³C isotope analyses, we have quantified two major C fluxes contributing to litter decomposition, at macroscopic and microscopic levels.     

Quantification of priming and CO2 emission sources following the application of different slurry particle size fractions to a grassland soil

The highest emissions of CO₂ from soils and most pronounced priming effect (PE) from soils generally occur immediately after slurry application. However, the influence of different particle size slurry fractions on net soil C respiration dynamics and PE has not been studied. Therefore, a slurry separation technique based on particle sizes was used in the present study. Six distinct fractions (>2000, 425-2000, 250-425, 150-250, 45-150, <45 μm) were generated from two dairy slurries (one from cows fed a predominantly maize silage diet and the other from cows fed a grass silage diet) were applied to soil. During the first days of the 332 days experiment, all slurry fraction amendments significantly increased soil CO₂ effluxes (by 2-8 times) compared to the non-amended control. The increased CO₂ emission rates had a negative relationship with slurry particle size, but its duration was positively correlated with slurry particle size. The percentage of the cumulative CO₂ emitted was only higher in the first 8 days in the finest slurry particle sizes (<150 μm). The proportion of slurry-derived C emitted as CO₂ 2 h after addition to soil varied between 29% and 100% of total emitted CO₂-C. Generally, the proportion of slurry-derived C emitted initially decreased rapidly in the <250 μm fractions, but decreased more slowly or even increased in the >250 μm fractions. The overall contribution of slurry C to total CO₂ emissions was higher in smaller slurry particle size treatments in the first days after application. The addition of the various slurry fractions to soil caused both significant positive and negative PEs on the soil organic matter mineralization. The timing and type (positive or negative) of PE depended on the slurry particle size. Clearly, farm based separation pre-treatment leading to two or more fractions with different particle sizes has also the potential to reduce or modify short-term CO₂ emissions immediately after slurry application to soil.     

Impacts of 22-year organic and inorganic N managements on soil organic C fractions in a maize field,northeast China

Impacts of 22-year organic and inorganic N managements on total organic carbon (TOC), water-soluble organic C (WSOC), microbial biomass C (MBC), particulate organic C (POC) and KMnO₄ oxidized organic C (KMnO₄-C) concentrations, C management index (CMI), and C storage in surface soil (0–20 cm) were investigated in a maize (Zea may L.) field experiment, Northeast China. The treatments included, CK: unfertilized control, M: organic manure (135 kg N ha^− 1 year^− 1), N: inorganic N fertilizer (135 kg N ha^− 1 year^− 1) and MN: combination of organic manure (67.5 kg N ha^− 1 year^− 1) and inorganic N fertilizer (67.5 kg N ha^− 1 year^− 1). TOC concentration and C storage were significantly increased under the M and MN treatments, but not under the inorganic N treatment. The organic treatments of M and MN were more effective in increasing WSOC, MBC, POC and KMnO₄-C concentrations and CMI than the N treatment. The M treatment was most effective for sequestrating SOC (10.6 Mg ha^− 1) and showed similar increase in degree of grain yield to the N and MN treatments, therefore it could be the best option for improving soil productivity and C storage in the maize cropping system.     

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.     

Characterizing the impact of diffusive and advective soil gas transport on the measurement and interpretation of the isotopic signal of soil respiration

By measuring the isotopic signature of soil respiration, we seek to learn the isotopic composition of the carbon respired in the soil (δ¹³C_R-s) so that we may draw inferences about ecosystem processes. Requisite to this goal is the need to understand how δ¹³C_R-s is affected by both contributions of multiple carbon sources to respiration and fractionation due to soil gas transport. In this study, we measured potential isotopic sources to determine their contributions to δ¹³C_R-s and we performed a series of experiments to investigate the impact of soil gas transport on δ¹³C_R-s estimates. The objectives of these experiments were to: i) compare estimates of δ¹³C_R-s derived from aboveground and belowground techniques, ii) evaluate the roles of diffusion and advection in a forest soil on the estimates of δ¹³C_R-s, and iii) determine the contribution of new and old carbon sources to δ¹³C_R-s for a Douglas-fir stand in the Pacific Northwest during our measurement period. We found a maximum difference of −2.36‰ between estimates of δ¹³C_R-s based on aboveground vs. belowground measurements; the aboveground estimate was enriched relative to the belowground estimate. Soil gas transport during the experiment was primarily by diffusion and the average belowground estimate of δ¹³C_R-s was enriched by 3.8-4.0‰ with respect to the source estimates from steady-state transport models. The affect of natural fluctuations in advective soil gas transport was little to non-existent; however, an advection-diffusion model was more accurate than a model based solely on diffusion in predicting the isotopic samples near the soil surface. Thus, estimates made from belowground gas samples will improve with an increase in samples near the soil surface. We measured a −1‰ difference in δ¹³C_R-s as a result of an experiment where advection was induced, a value which may represent an upper limit in fractionation due to advective gas transport in forest ecosystems. We found that aboveground measurements of δ¹³C_R-s may be particularly susceptible to atmospheric incursion, which may produce estimates that are enriched in ¹³C. The partitioning results attributed 69-98% of soil respiration to a source with a highly depleted isotopic signature similar to that of water-soluble carbon from foliage measured at our site.     

Differences in isotopic fractionation of nitrogen in water-saturated and unsaturated soils

Temporal variations in δ¹⁵N of NH₄⁺ and NO₃⁻ in water-saturated and unsaturated soils were examined in a laboratory incubation study. Ammonium sulfate (δ¹⁵N=−2.6‰) was added to 25 g samples of soil at concentrations of 160 mg N kg⁻¹. Soils were then incubated under unsaturated (50% of water holding capacity at saturation, WHC) or saturated (100% of WHC) water conditions for 7 and 36 d, respectively. During 7 d incubation of unsaturated soil, the NH₄⁺-N concentration decreased from 164.8 to 34.4 mg kg⁻¹, and the δ¹⁵N of NH₄⁺ increased from −0.4 to +57.2‰ through nitrification, as evidenced by corresponding increase in NO₃⁻-N concentration and lower δ¹⁵N of NO₃⁻ (product) than that of NH₄⁺ (substrate) at each sampling time. In saturated soil, the concentration of NH₄⁺-N decreased gradually from 162.4 to 24.2 mg kg⁻¹, and the δ¹⁵N values increased from +0.8 to +21.0‰ during 36 d incubation. However, increase in NO₃⁻ concentration was not observed due to loss of NO₃⁻ through concurrent denitrification in anaerobic sites. The apparent isotopic fractionation factors (α_s/p) associated with decrease in NH₄⁺ concentration were 1.04 and 1.01 in unsaturated and saturated soils, respectively. Since nitrification is likely to introduce greater isotope fractionation than microbial immobilization, the higher value for unsaturated soil probably reflected faster nitrification under aerobic conditions. The lower value for saturated soil suggests that immobilization and subsequent remineralization of NH₄⁺ were relatively more dominant than nitrification under the anaerobic conditions.     

Natural N abundances of inorganic nitrogen in soil treated with fertilizer and compost under changing soil moisture regimes

This study was conducted to examine whether the applications of N-inputs (compost and fertilizer) having different N isotopic compositions (δ¹⁵N) produce isotopically different inorganic-N and to investigate the effect of soil moisture regimes on the temporal variations in the δ¹⁵N of inorganic-N in soils. To do so, the temporal variations in the concentrations and the δ¹⁵N of NH₄⁺ and NO₃⁻ in soils treated with two levels (0 and 150 mg N kg⁻¹) of ammonium sulfate (δ¹⁵N=−2.3‰) and compost (+13.9‰) during a 10-week incubation were compared by changing soil moisture regime after 6 weeks either from saturated to unsaturated conditions or vice versa. Another incubation study using ¹⁵N-labeled ammonium sulfate (3.05 ¹⁵N atom%) was conducted to estimate the rates of nitrification and denitrification with a numerical model FLUAZ. The δ¹⁵N values of NH₄⁺ and NO₃⁻ were greatly affected by the availability of substrate for each of the nitrification and denitrification processes and the soil moisture status that affects the relative predominance between the two processes. Under saturated conditions for 6 weeks, the δ¹⁵N of NH₄⁺ in soils treated with fertilizer progressively increased from +2.9‰ at 0.5 week to +18.9‰ at 6 weeks due to nitrification. During the same period, NO₃⁻ concentrations were consistently low and the corresponding δ¹⁵N increased from +16.3 to +39.2‰ through denitrification. Under subsequent water-unsaturated conditions, the NO₃⁻ concentrations increased through nitrification, which resulted in the decrease in the δ¹⁵N of NO₃⁻. In soils, which were unsaturated for the first 6-weeks incubation, the δ¹⁵N of NH₄⁺ increased sharply at 0.5 week due to fast nitrification. On the other hand, the δ¹⁵N of NO₃⁻ showed the lowest value at 0.5 week due to incomplete nitrification, but after a subsequence increase, they remained stable while nitrification and denitrification were negligible between 1 and 6 weeks. Changing to saturated conditions after the initial 6-weeks incubation, however, increased the δ¹⁵N of NO₃⁻ progressively with a concurrent decrease in NO₃⁻ concentration through denitrification. The differences in δ¹⁵N of NO⁻₃ between compost and fertilizer treatments were consistent throughout the incubation period. The δ¹⁵N of NO₃⁻ increased with the addition of compost (range: +13.0 to +35.4‰), but decreased with the addition of fertilizer (−10.8 to +11.4‰), thus resulting in intermediate values in soils receiving both fertilizer and compost (−3.5 to +20.3‰). Therefore, such differences in δ¹⁵N of NO₃⁻ observed in this study suggest a possibility that the δ¹⁵N of upland-grown plants receiving compost would be higher than those treated with fertilizer because NO₃⁻ is the most abundant N for plant uptake in upland soils.     

Savanna-derived organic matter remaining in arable soils of the South African Highveld long-term mixed cropping: Evidence from C and N natural abundance

Sustainable agriculture requires the formation of new humus from the crops. We utilized ¹³C and ¹⁵N signatures of soil organic matter to assess how rapidly wheat/maize cropping contributed to the humus formation in coarse-textured savanna soils of the South African Highveld. Composite samples were taken from the top 20 cm of soils (Plinthustalfs) cropped for lengths of time varying from 0 to 98 years, after conversion from native grassland savanna (C4). We performed natural ¹³C and ¹⁵N abundance measurements on bulk and particle-size fractions. The bulk soil δ¹³C values steadily decreased from −14.6 in (C4 dominated) grassland to −16.5‰ after 90 years of arable cropping. This δ¹³C shift was attributable to increasing replacement of savanna-derived C by wheat crop (C3) C which dominated over maize (C4) inputs. After calculating the annual C input from the crop yields and the output from literature data, by using a stepwise C replacement model, we were able to correct the soil δ¹³C data for the irregular maize inputs for a period of about one century. Within 90 years of cropping 41-89% of the remaining soil organic matter was crop-derived in the three studied agroecosystems. The surface soil C stocks after 90 years of the wheat/maize crop rotation could accurately be described with the Rothamsted Carbon Model, but modelled C inputs to the soil were very low. The coarse sand fraction reflected temporal fluctuations in ¹³C of the last C₃ or C₄ cropping and the silt fraction evidenced selective erosion loss of old savanna-derived C. Bulk soil ¹⁵N did not change with increasing cropping length. Decreasing δ¹⁵N values caused by fertilizer N inputs with prolonged arable cropping were only detected for the coarse sand fraction. This indicated that the present N fertilization was not retained in stable soil C pool. Clearly, conventional cropping practices on the South African highlands neither contribute to the preservation of old savanna C and N, nor the effective humus reformation by the crops.     

Nitrogen fixation by biological soil crusts and heterotrophic bacteria in an intact Mojave Desert ecosystem with elevated CO2 and added soil carbon

Fixation of N by biological soil crusts and free-living heterotrophic soil microbes provides a significant proportion of ecosystem N in arid lands. To gain a better understanding of how elevated CO₂ may affect N₂-fixation in aridland ecosystems, we measured C₂H₂ reduction as a proxy for nitrogenase activity in biological soil crusts for 2 yr, and in soils either with or without dextrose-C additions for 1 yr, in an intact Mojave Desert ecosystem exposed to elevated CO₂. We also measured crust and soil δ¹⁵N and total N to assess changes in N sources, and δ¹³C of crusts to determine a functional shift in crust species, with elevated CO₂. The mean rate of C₂H₂ reduction by biological soil crusts was 76.9±5.6 μmol C₂H₄ m⁻² h⁻¹. There was no significant CO₂ effect, but crusts from plant interspaces showed high variability in nitrogenase activity with elevated CO₂. Additions of dextrose-C had a positive effect on rates of C₂H₂ reduction in soil. There was no elevated CO₂ effect on soil nitrogenase activity. Plant cover affected soil response to C addition, with the largest response in plant interspaces. The mean rate of C₂H₂ reduction in soils either with or without C additions were 8.5±3.6 μmol C₂H₄ m⁻² h⁻¹ and 4.8±2.1 μmol m⁻² h⁻¹, respectively. Crust and soil δ¹⁵N and δ¹³C values were not affected by CO₂ treatment, but did show an effect of cover type. Crust and soil samples in plant interspaces had the lowest values for both measurements. Analysis of soil and crust [N] and δ¹⁵N data with the Rayleigh distillation model suggests that any plant community changes with elevated CO₂ and concomitant changes in litter composition likely will overwhelm any physiological changes in N₂-fixation.