Relationships between C and N availability, substrate age, and natural abundance C and N signatures of soil microbial biomass in a semiarid climate

Soil microbial organisms are central to carbon (C) and nitrogen (N) transformations in soils, yet not much is known about the stable isotope composition of these essential regulators of element cycles. We investigated the relationship between C and N availability and stable C and N isotope composition of soil microbial biomass across a three million year old semiarid substrate age gradient in northern Arizona. The δ¹⁵N of soil microbial biomass was on average 7.2‰ higher than that of soil total N for all substrate ages and 1.6‰ higher than that of extractable N, but not significantly different for the youngest and oldest sites. Microbial ¹⁵N enrichment relative to soil extractable and total N was low at the youngest site, increased to a maximum after 55,000 years, and then decreased slightly with age. The degree of ¹⁵N enrichment of microbial biomass correlated negatively with the C:N mass ratio of the soil extractable pool. The δ¹³C signature of soil microbial biomass was 1.4‰ and 4.6‰ enriched relative to that of soil total and extractable pools respectively and showed significant differences between sites. However, microbial ¹³C enrichment was unrelated to measures of C and N availability. Our results confirm that ¹⁵N, but not ¹³C enrichment of soil microbial biomass reflects changes in C and N availability and N processing during long-term ecosystem development.     

Dissipation of bacterially derived C and N through the meso- and macrofauna of a grassland soil

Feeding relationships between organisms may be determined by observations of behaviour in manipulative experiments or, as in more recent times, by the use of stable isotope labelling to trace the passage of ¹³C and ¹⁵N through food webs. Here we introduce living bacteria, labelled with both ¹³C and ¹⁵N into intact soil cores to understand further the movement of bacterially sourced C and N into the meso- and macrofauna of a grassland soil. We found that these groups showed a range of isotope levels which relate to their feeding strategies. Some had no label (e.g. dipterous larvae), whilst others were highly labelled which may indicate a preference for the added bacteria. This latter group included Collembola, generally perceived as being predominantly fungal feeders. This work describes a novel technique which has the potential to provide critical information about the dissipation of bacterially derived C and N through the soil food web.     

Extractability of labelled microbial biomass N by electro-ultrafiltration and CaCl2 extraction

A laboratory soil incubation and a pot experiment with ryegrass were carried out in order to examine the extractability of microbial biomass N by using either 10-mM CaCl₂ extraction or the electro-ultrafiltration (EUF) method. The aim of the experiment was to test the hypothesis whether the organic N (Norg) extracted by EUF or CaCl₂ from dried soil samples represents a part of the microbial biomass. For the laboratory incubation a ¹⁵N-labelled Escherichia coli suspension was mixed with the soil. For the pot experiment a suspension of ¹⁵N-labelled bacteria was applied which had previously been isolated from the soil used. Soil samples of both treatments, with and without applied bacterial suspension, were extracted by EUF and CaCl₂. The extractability of applied microbial biomass was estimated from the difference in extractable Norg between the two treatments. In addition, the N isotopic composition in the upper plant matter, in the soil, and in organic and inorganic N fractions of EUF and CaCl₂ extracts was analysed. Both experiments showed that the applied microbial biomass was highly accessible to mineralization and thus represented potentially mineralizable N. However, this mineralizable N was not extractable by CaCl₂ or by the EUF method. It was, therefore, concluded that the organic N released on soil drying and which was thus extractable was derived from the non-biomass soil organic matter. The result suggests that both extraction methods may provide a suitable index for mineralizable N only in cases where the decomposable organic substrates are derived mainly from sources other than the living soil biota.Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday     

Plant nitrate use in deciduous woodland: the relationship between leaf N, N natural abundance of forbs and soil N mineralisation

Our aim was to study whether the in situ natural abundance ¹⁵N (δ¹⁵N)-values and N concentration of understory plants were correlated with the form and amount of mineral N available in the soil. Also to determine whether such differences were related to earlier demonstrations of differences in biomass increase in the same species exposed to nutrient solutions with both and or to alone. Several studies show that the δ¹⁵N of in soil solution generally is isotopically lighter than the δ¹⁵N of due to fractionation during nitrification. Hence, it is reasonable to assume that plant species benefiting from in ecosystems without significant leaching or denitrification have lower δ¹⁵N-values in their tissues than species growing equally well, or better, on We studied the δ¹⁵N of six understory species in oak woodlands in southern Sweden at 12 sites which varied fivefold in potential net N mineralisation rate The species decreased in benefit from in the following order: Geum urbanum, Aegopodium podagraria, Milium effusum, Convallaria majalis, Deschampsia flexuosa and Poa nemoralis. Four or five species demonstrated a negative correlation between and leaf δ¹⁵N and a positive correlation between and leaf N concentration. In wide contrast, only D. flexuosa, which grows on soils with little nitrification, showed a positive correlation between and the leaf N concentration and δ¹⁵N-value. Furthermore, δ¹⁵N of plants from the field and previously obtained indices of hydroponic growth on relative to were closely correlated at the species level. We conclude that δ¹⁵N may serve as a comparative index of uptake of among understory species, preferably in combination with other indices of N availability. The use of δ¹⁵N needs careful consideration of known restrictions of method, soils and plants.     

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.     

Three-source partitioning of CO2 efflux from soil planted with maize by C natural abundance fails due to inactive microbial biomass

A theoretical approach to the partitioning of carbon dioxide (CO₂) efflux from soil with a C₃ vegetation history planted with maize (Zea mays), a C₄ plant, into three sources, root respiration (RR), rhizomicrobial respiration (RMR), and microbial soil organic matter (SOM) decomposition (SOMD), was examined. The δ¹³C values of SOM, roots, microbial biomass, and total CO₂ efflux were measured during a 40-day growing period. A three-source isotopic mass balance based on the measured δ¹³C values and on assumptions made in other studies showed that RR, RMR, and SOMD amounted to 91%, 4%, and 5%, respectively. Two assumptions were thoroughly examined in a sensitivity analysis: the absence of ¹³C fractionation and the conformity of δ¹³C of microbial CO₂ and that of microbial biomass. This approach strongly overestimated RR and underestimated RMR and microbial SOMD. CO₂ efflux from unplanted soil was enriched in ¹³C by 2.0‰ compared to microbial biomass. The consideration of this ¹³C fractionation in the mass balance equation changed the proportions of RR and RMR by only 4% and did not affect SOMD. A calculated δ¹³C value of microbial CO₂ by a mass balance equation including active and inactive parts of microbial biomass was used to adjust a hypothetical below-ground CO₂ partitioning to the measured and literature data. The active microbial biomass in the rhizosphere amounted to 37% to achieve an appropriate ratio between RR and RMR compared to measured data. Therefore, the three-source partitioning approach failed due to a low active portion of microbial biomass, which is the main microbial CO₂ source controlling the δ¹³C value of total microbial biomass. Since fumigation-extraction reflects total microbial biomass, its δ¹³C value was unsuitable to predict δ¹³C of released microbial CO₂ after a C₃-C₄ vegetation change. The second adjustment to the CO₂ partitioning results in the literature showed that at least 71% of the active microbial biomass utilizing maize rhizodeposits would be necessary to achieve that proportion between RR and RMR observed by other approaches based on ¹⁴C labelling. The method for partitioning total below-ground CO₂ efflux into three sources using a natural ¹³C labelling technique failed due to the small proportion of active microbial biomass in the rhizosphere. This small active fraction led to a discrepancy between δ¹³C values of microbial biomass and of microbially respired CO₂.     

Natural N abundance of plant and soil inorganic-N as evidence for over-fertilization with compost

To test the hypothesis that N isotope composition can be used as evidence of excessive compost application, we measured variation in patterns of N concentrations and corresponding δ¹⁵N values of plants and soil after compost application. To do so, a pot experiment with Chinese cabbage (Brassica campestris L. cv. Maeryok) was conducted for 42 days. Compost was applied at rates of 0 (SC0), 500 (SC1), 1000 (SC2), and 1500 mg N kg⁻¹ soil (SC3). Plant-N uptake linearly increased with compost application (r² = 0.956, P < 0.05) with an uptake efficiency of 76 g N kg⁻¹ of compost-N at 42 days after application, while dry-mass accumulation did not show such linear increases. Net N mineralized from compost-N increased linearly (r² = 0.998, P < 0.01) with a slope of 122 g N kg⁻¹ of compost-N. Plant-δ¹⁵N increased curvilinearly with increasing compost application, but this increase was insignificant between SC2 and SC3 treatments. The δ¹⁵N of soil inorganic-N (particularly NO₃⁻-N) increased with compost application. We found that plant-δ¹⁵N reflected the N isotope signal of soil NO₃⁻-N at each measurement during plant growth, and that δ¹⁵N of inner leaves and soil NO₃⁻-N was similar when initial NO₃⁻ in the compost was abundant. Therefore, we concluded that δ¹⁵N of whole plant (more obviously in newer plant parts) and soil NO₃⁻-N could reveal whether compost application was excessive, suggesting a possible use of δ¹⁵N in plants and soil as evidence of excess compost application.     

土壤C/N对苹果植株生长及氮素利用的影响

土壤C/N是土壤氮素循环的重要影响因素。本研究以2年生"富士"/平邑甜茶为试验材料, 应用¹⁵N示踪技术研究了不同土壤C/N[6.21(CK)、10、15、20、25、30、35和40]对苹果植株生长及氮素利用和损失的影响。结果表明: 随着土壤C/N比值的逐渐增大, 苹果新梢长度和植株鲜重均呈先升高后降低的变化趋势, C/N=15、20和25的3个处理苹果新梢长度和植株鲜重最大, 三者间无显著差异, 但均显著高于其他处理。不同C/N处理间植株15N利用率存在差异, 土壤C/N=25时, 植株¹⁵N利用率最大, 为22.87%, 与C/N=20的处理间无显著差异, 但两者均显著高于其他处理; 土壤C/N=40时, 植株¹⁵N利用率最低, 仅为15.43%, 低于CK处理的16.65%。土壤C/N处于15~25时, 植株吸收的氮素来自于肥料氮的比例较高; 而土壤C/N较低(<15)或太高(>25)时, 植株吸收的氮素来自于土壤氮的比例较高。土壤氮素残留量随土壤C/N的增大逐渐增加, C/N=40处理的土壤氮素残留量是CK的1.32倍。随着土壤C/N比值的逐渐增大, 肥料氮损失量呈先减少后增加的变化趋势, 以C/N=25时最少, 仅为施氮量的49.87%, 而对照最大, 为61.54%。因此, 综合土壤C/N对苹果植株生长及氮素平衡状况来看, 土壤C/N为15~25时, 能促进植株的生长发育, 降低氮肥损失, 提高肥料利用率。     

Natural 15N and 13C abundance in Andisols influenced by long-term fertilization management in Japan

Two field experiments were conducted on Andisols in Japan to evaluate the changes in the natural ¹⁵N and ¹³C abundance in the soil profile and to determine whether the values of δ¹⁵N could be used as an indicator of fertilizer sources or fertilizer fate. The 6-year experiment conducted at the National Agricultural Research Center (NARC) consisted of the following treatments: application of swine compost (COMPOST), slow-release nitrogen fertilizer (SRNF), readily available nitrogen fertilizer (RANF), and absence of fertilization (CONTROL). Experimental plots located at the Nippon Agricultural Research Institute (NARI) received cattle compost at different rates for 12 years; a forest soil at this site was sampled for comparison. Swine compost application led to a considerable change in the δ¹⁵N distribution pattern in the soil profile, with the highest δ¹⁵N values recorded in the top 20 cm layers of the COMPOST plot, decreasing in the sequence of CONTROL >- RANF > SRNF, mainly due to the relatively high δ¹⁵N value of swine compost and its subsequent decomposition. In contrast, SRNF application resulted in the lowest δ¹⁵N values in soil, indicating the presence of negligible nitrogen losses relative to input and low nitrogen cycling rates. Values of δ¹⁵N increased with compost application rates at NARI. In the leachate collected at 1-m depth, the δ¹⁵N values decreased in the sequence of COMPOST > RANF ≥ CONTROL > SRNF. The δ¹³C values in soil peaked in the 40–60 cm layers for all the fertilizers. The δ¹³C value was lowest in forest soil due to the presence of plant residues in soil organic matter. These results indicated that the δ¹⁵N values in the upper soil layers or leachate may enable to detect pollution sources of organic or inorganic nitrogen qualitatively in Andisols.     

Compartmentalization of the soil animal food web as indicated by dual analysis of stable isotope ratios (N/N and C/C)

The soil animal food web has become a focus of recent ecological research but trophic relationships still remain enigmatic for many taxa. Analysis of stable isotope ratios of N and C provides a powerful tool for disentangling food web structure. In this study, animals, roots, soil and litter material from a temperate deciduous forest were analysed. The combined measurement of δ¹⁵N and δ¹³C provided insights into the compartmentalization of the soil animal food web. Leaf litter feeders were separated from animals relying mainly on recent belowground carbon resources and from animals feeding on older carbon. The trophic pathway of leaf litter-feeding species appears to be a dead end, presumably because leaf litter feeders (mainly diplopods and oribatid mites) are unavailable to predators due to large size and/or strong sclerotization. Endogeic earthworms that rely on older carbon also appear to exist in predator-free space. The data suggest that the largest trophic compartment constitutes of ectomycorrhizal feeders and their predators. Additionally, there is a smaller trophic compartment consisting of predators likely feeding on enchytraeids and potentially nematodes.     

Enrichment of natural 15N abundance in frankia-infected nodules

Abstract

The enrichment of ¹⁵N in the nodules of some N₂-fixing leguminous plants is an interesting finding (Shearer et al. 1982). The extent of ¹⁵N enrichment differed depending on the plant species (Shearer et al. 1982; Yoneyama 1987) and bacterial strains (Steele et al. 1983), and in soybeans it was apparently related to the nitrogen fixation efficiency (Shearer et al. 1984)     

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.     

Organic matter turnover in soil physical fractions following woody plant invasion of grassland: Evidence from natural C and N

Soil physical structure causes differential accessibility of soil organic carbon (SOC) to decomposer organisms and is an important determinant of SOC storage and turnover. Techniques for physical fractionation of soil organic matter in conjunction with isotopic analyses (δ¹³C, δ¹⁵N) of those soil fractions have been used previously to (a) determine where organic C is stored relative to aggregate structure, (b) identify sources of SOC, (c) quantify turnover rates of SOC in specific soil fractions, and (d) evaluate organic matter quality. We used these two complementary approaches to characterize soil C storage and dynamics in the Rio Grande Plains of southern Texas where C₃ trees/shrubs (δ¹³C=−27‰) have largely replaced C₄ grasslands (δ¹³C=−14‰) over the past 100-200 years. Using a chronosequence approach, soils were collected from remnant grasslands (Time 0) and from woody plant stands ranging in age from 10 to 130 years. We separated soil organic matter into specific size/density fractions and determined their C and N concentrations and natural δ¹³C and δ¹⁵N values. Mean residence times (MRTs) of soil fractions were calculated based on changes in their δ¹³C with time after woody encroachment. The shortest MRTs (average=30 years) were associated with all particulate organic matter (POM) fractions not protected within aggregates. Fine POM (53-250 μm) within macro- and microaggregates was relatively more protected from decay, with an average MRT of 60 years. All silt+clay fractions had the longest MRTs (average=360 years) regardless of whether they were found inside or outside of aggregate structure. δ¹⁵N values of soil physical fractions were positively correlated with MRTs of the same fractions, suggesting that higher δ¹⁵N values reflect an increased degree of humification. Increased soil C and N pools in wooded areas were due to both the retention of older C₄-derived organic matter by protection within microaggregates and association with silt+clay, and the accumulation of new C₃-derived organic matter in macroaggregates and POM fractions.     

Near infrared reflectance spectroscopy for determination of organic matter fractions including microbial biomass in coniferous forest soils

The objective of this work was to investigate the usefulness of near infrared reflectance spectroscopy (NIRS) in determining some C and N fractions of soils: labile compounds, microbial biomass, compounds derived from added ¹³C- and ¹⁵N-labelled straw. Soil samples were obtained from a previous experiment where soils were labelled by addition of ¹³C- and ¹⁵N-labelled wheat straw and incubated in coniferous forests in northern Sweden (64-60°N) and south France (43°N). The incubation lasted three years with 7-9 samplings at regular time steps and four replicates at each sampling (204 samples). Samples were scanned using a near infrared reflectance spectrophotometer (NIRSystem 6500). Calibrations were obtained by using a modified partial least squares regression technique with reference data on total C and N, ¹³C, ¹⁵N, control extract-C, -N, -¹³C and -¹⁵N, fumigated extract-C, -N, -¹³C and -¹⁵N, biomass-C, -N, -¹³C and -¹⁵N contents. Mathematical treatments of the absorbance data were first or second derivative with a gap from 4 to 10 nm. The standard error of calibration (SEC)-to-standard deviation of the reference measurements ratio was ≤0.2 for 10 models, namely total C and N, ¹³C, ¹⁵N, control extract-C, fumigated extract-C and -N, biomass-C and -N and biomass-¹⁵N models and therefore considered as very good. With an R²=0.955, the fumigated extract-¹⁵N model is also good. The standard error of performance calculated on the independent set of data and SEC were within 20% of each other for all the best equations except for the biomass-¹⁵N model. The ability of NIRS to detect ¹³C and ¹⁵N in total C and N and in the extracts is noteworthy, not because of its predictive function that is not really of interest in this case, but because it indicates that the spectra kept the signature of the properties of the organic matter derived from the straw even after two- or three-year decomposition. The incorporation of the ¹³C in the biomass was less well predicted than that of the ¹⁵N. This could indicate that the biomass derived from the straw was characterised by a particular protein or amino acid composition compared to the total biomass that includes a large proportion of dormant micro-organisms. The predictive ability of NIRS for microbial biomass-C and -N is particularly interesting because the conventional analyses are time consuming. In addition, NIRS allows detecting analytical errors.     

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.     

Black carbon decomposition and incorporation into soil microbial biomass estimated by C labeling

Incomplete combustion of organics such as vegetation or fossil fuel led to accumulation of charred products in the upper soil horizon. Such charred products, frequently called pyrogenic carbon or black carbon (BC), may act as an important long-term carbon (C) sink because its microbial decomposition and chemical transformation is probably very slow. Direct estimations of BC decomposition rates are absent because the BC content changes are too small for any relevant experimental period. Estimations based on CO₂ efflux are also unsuitable because the contribution of BC to CO₂ is too small compared to soil organic matter (SOM) and other sources.We produced BC by charring ¹⁴C labeled residues of perennial ryegrass (Lolium perenne). We then incubated this ¹⁴C labeled BC in Ah of a Haplic Luvisol soil originated from loess or in loess for 3.2 years. The decomposition rates of BC were estimated based on ¹⁴CO₂ sampled 44 times during the 3.2 years incubation period (1181 days). Additionally we introduced five repeated treatments with either 1) addition of glucose as an energy source for microorganisms to initiate cometabolic BC decomposition or 2) intensive mixing of the soil to check the effect of mechanical disturbance of aggregates on BC decomposition. Black carbon addition amounting to 20% of C_org of the soil or 200% of C_org of loess did not change total CO₂ efflux from the soil and slightly decreased it from the loess. This shows a very low BC contribution to recent CO₂ fluxes. The decomposition rates of BC calculated based on ¹⁴C in CO₂ were similar in soil and in loess and amounted to 1.36 10⁻⁵ d⁻¹ (=1.36 10⁻³% d⁻¹). This corresponds to a decomposition of about 0.5% BC per year under optimal conditions. Considering about 10 times slower decomposition of BC under natural conditions, the mean residence time (MRT) of BC is about 2000 years, and the half-life is about 1400 years. Considering the short duration of the incubation and the typical decreasing decomposition rates with time, we conclude that the MRT of BC in soils is in the range of millennia.The strong increase in BC decomposition rates (up to 6 times) after adding glucose and the decrease of this stimulation after 2 weeks in the soil (and after 3 months in loess) allowed us to conclude cometabolic BC decomposition. This was supported by higher stimulation of BC decomposition by glucose addition compared to mechanical disturbance as well as higher glucose effects in loess compared to the soil. The effect of mechanical disturbance was over within 2 weeks. The incorporation of BC into microorganisms (fumigation/extraction) after 624 days of incubation amounted to 2.6 and 1.5% of ¹⁴C input into soil and loess, respectively. The amount of BC in dissolved organic carbon (DOC) was below the detection limit (<0.01%) showing no BC decomposition products in water leached from the soil.We conclude that applying ¹⁴C labeled BC opens new ways for very sensitive tracing of BC transformation products in released CO₂, microbial biomass, DOC, and SOM pools with various properties.     

Comparison of 15N natural abundance and difference methods for estimating N2 fixation of soybeans grown in a tropical soil

Abstract

A study was carried out to compare the difference or N-yield method with the ¹⁵N natural abundance method for the estimation of the fractional contribution of biological N₂ fixation in the different plant parts of nodulating and non-nodulating isolines of soybeans. The results indicated that the δ¹⁵N values of most plant parts of soybeans were significantly lower (p<0.05) in the nodulating than in the non-nodulating isoline. However, in the case of the root+nodule component, the δ¹⁵N value was higher in the nodulating than in the non-nodulating isoline possibly due to isotopic discrimination of ¹⁵N over ¹⁴N which may have occurred in the nodules. Inoculation of soybeans with the Bradyrhizobium japonicum strain CB 1809 increased significantly (p<0.05) the δ¹⁵N value of the root+nodule component implying that the effectiveness of the soybean-rhizobium symbiosis had increased by inoculation.

Percentage of plant N derived from atmospheric N₂ fixation (%Ndfa) estimated by the ¹⁵N natural abundance method was highly correlated (r=0.762, p<0.01) with that by the difference or N-yield method and the differences between the two methods were not statistically significant. The agreement between the two methods was closer at maturity than at the early reproductive stage.

The %Ndfa obtained by the difference method ranged from 48.4 to 92.6% whereas the %Ndfa obtained by the ¹⁵N natural abundance method ranged from 43.2 to 92.4% in the different plant parts. Based on the ¹⁵N natural abundance method, approximately 15% of the N in pod, shoot, grain, and shell was derived from the soil but in the case of stover, this fraction was about 55%.     

Variation in 15N natural abundance of soil, humic fractions and plant materials in a disturbed and an undisturbed grassland

The natural abundance of δ¹⁵N in disturbed and undisturbed pasture soils was examined. From the disturbed soil, the top 10 cm of the profile was examined and the soil split into fractions based on particle size. Plant shoot and root material contained similar low enrichments in ¹⁵N, whereas recently deposited shoot residues were highly enriched. Differences between the soil fractions in observed total N did not reflect similar ¹⁵N variation. However, the enrichment of humic material extracted from the largest soil fraction was considerably lower in ¹⁵N relative to that from the smaller fractions. The complexity of the humic material from the larger fractions was less according to the E ₄ /E ₆ ratio. Analysis of the profile from the undisturbed soil showed increasing ¹⁵N enrichment with depth which corresponded well with visible soil horizons and showed an inverse relationship with total soil N. This ¹⁵N enrichment was mirrored by the enrichment in humic materials down the profile and also corresponded with an increasing chemical complexity as shown by the E ₄ /E ₆ ratio. Received: 15 March 1996     

Carbon dynamics determined by natural C abundance in microcosm experiments with soils from long-term maize and rye monocultures

Understanding carbon dynamics in soil is the key to managing soil organic matter. Our objective was to quantify the carbon dynamics in microcosm experiments with soils from long-term rye and maize monocultures using natural ¹³C abundance. Microcosms with undisturbed soil columns from the surface soil (0-25 cm) and subsoil (25-50 cm) of plots cultivated with rye (C₃-plant) since 1878 and maize (C₄-plant) since 1961 with and without NPK fertilization from the long-term experiment ‘Ewiger Roggen’ in Halle, Germany, were incubated for 230 days at 8 °C and irrigated with 2 mm 10⁻² M CaCl₂ per day. Younger, C₄-derived and older, C₃-derived percentages of soil organic carbon (SOC), dissolved organic carbon (DOC), microbial biomass (C_mic) and CO₂ from heterothropic respiration were determined by natural ¹³C abundance. The percentage of maize-derived carbon was highest in CO₂ (42-79%), followed by C_mic (23-46%), DOC (5-30%) and SOC (5-14%) in the surface soils and subsoils of the maize plots. The percentage of maize-derived C was higher for the NPK plot than for the unfertilized plot and higher for the surface soils than for the subsoils. Specific production rates of DOC, CO₂-C and C_mic from the maize-derived SOC were 0.06-0.08% for DOC, 1.6-2.6% for CO₂-C and 1.9-2.7% for C_mic, respectively, and specific production rates from rye-derived SOC of the continuous maize plot were 0.03-0.05% for DOC, 0.1-0.2% for CO₂-C and 0.3-0.5% for C_mic. NPK fertilization did not affect the specific production rates. Strong correlations were found between C₄-derived C_mic and C₄-derived SOC, DOC and CO₂-C (r≥0.90), whereas the relationship between C₃-derived C_mic and C₃-derived SOC, DOC and CO₂-C was not as pronounced (r≤0.67). The results stress the different importance of former (older than 40 years) and recent (younger than 40 years) litter C inputs for the formation of different C pools in the soil.     

Cultivation effects on biochemical properties, C storage and N natural abundance in the 0–5 cm layer of an acidic soil from temperate humid zone

Changes in some soil chemical, including ¹⁵N values, and biochemical properties (microbial C, FDA hydrolysis, glucosidase and urease activities) due to two tillage systems, conventional tillage (CT) and no-tillage (NT), were evaluated in an acid soil from temperate humid zone (NW of Spain) and compared with values obtained for a reference forest soil. The results showed that in the surface layer (0–5 cm depth) tillage tended to increase soil pH and to decrease organic matter levels and microbial biomass and activity values. The data also indicated that 8 years of NT, compared to CT, resulted in greater organic matter content and increased microbial biomass and activity, the changes being more pronounced for the microbial properties. Adoption of NT resulted in an increase of soil C storage of 1.24 Mg C ha⁻¹ year⁻¹ with regard to CT. The suitability of ¹⁵N as a potential tracer of land-use in this acid soil was also confirmed.