Carbohydrate and amino acid composition of dissolved organic matter leached from soil

Low molecular weight organic substances (LMWOS) in soil and soil solution include mainly amino acids, carboxylic acids, and carbohydrates. Due to their high bioavailability they play a crucial role in the cycles of C and nutrients in soils. The variety of soil processes that involve LMWOS requires identifying their composition to elucidate reactions and transformations. In most studies, LMWOS are extracted under artificial conditions, e.g. batch experiments, which may overestimate the actual concentrations. This study measures the composition of carbohydrates and amino acids in solution of a Haplic Luvisol leached in a column experiment. A combined system for simultaneous leaching and blowout of CO₂ was used to estimate LMWOS decomposition. ¹⁴C-labeled glucose was added as a highly sensitive tracer to control the efficiency of the LMWOS extraction by leaching and to estimate LMWOS decomposition during leaching. High performance liquid chromatography (HPLC), optimized for soil extracts, was used to analyze LMWOS composition. For HPLC optimization, different preparations of leached solutions (filtration vs. centrifugation, and drying vs. no-drying) were compared. For sugar determination, drying had no influence on the solution concentrations. In contrast, amino acid concentrations significantly decreased by drying LMWOS eluted substances. Combining the HPLC identification of eluted substances with ¹⁴C tracer application revealed that about 5% of the glucose could be leached unchanged within 786 min (13.1 h), whereas about 84% remained in the soil, 9% were decomposed to CO₂, and 2% were transformed to other LMWOS and recovered in the soil solution. The total amino acid concentration (TAC) in soil solution was about 8.2 μmol l⁻¹, dominated by alanine (14.4% of TAC), glycine (13.4%), glutamic acid (9.9%), serine (9.4%), and leucine (9.3%). The total carbohydrate concentration was about 2.4 μM, dominated by glucose (29.9%), glucuronic acid (26.8%), and galacturonic acid (17.3%). Ratios of hexoses to pentoses, amino sugars glucosamine to galactosamine, and neutral sugars to uronic acids were determined. All three parameters pointed to the dominant influence of plants as the source of LMWOS in the leached soil solution. Within the small contribution of microorganisms, bacteria dominated over fungi. These used biomarker ratios as well as LMWOS concentrations differed widely from the ones obtained with conventional batch extraction. More research is necessary to evaluate the application of these biomarkers to soil solutions.     

Fate of low molecular weight organic substances in an arable soil: From microbial uptake to utilisation and stabilisation

Microbial uptake and utilisation are the main transformation pathways of low molecular weight organic substances (LMWOS) in soil, but details on transformations are strongly limited. As various LMWOS classes enter biochemical cycles at different steps, we hypothesize that the percentage of their carbon (C) incorporation into microbial biomass and consequently stabilisation in soil are different.Representatives of the three main groups of LMWOS: amino acids (alanine, glutamate), sugars (glucose, ribose) and carboxylic acids (acetate, palmitate) – were applied at naturally-occurring concentrations into a loamy arable Luvisol in a field experiment. Incorporation of ¹³C from these LMWOS into extractable microbial biomass (EMB) and into phospholipid fatty acids (PLFAs) was investigated 3 d and 10 d after application. The microbial utilisation of LMWOS for cell membrane construction was estimated by replacement of PLFA-C with ¹³C.35–80% of initially applied LMWOS-¹³C was still present in the composition of soil organic matter after 10 days of experiment, with 10–24% of ¹³C incorporation into EMB at day three and 1–15% at day 10. Maximal incorporation of ¹³C into EMB was observed from sugars and the least from amino acids. Strong differences in microbial utilisation between LMWOS were observed mainly at day 10. Thus, despite similar initial rapid uptake by microorganisms, further metabolism within microbial cells accounts for the specific fate of C from various LMWOS in soils.¹³C from each LMWOS was incorporated into each PLFA. This reflects the ubiquitous utilisation of all LMWOS by all functional microbial groups. The preferential incorporation of palmitate into PLFAs reflects its role as a direct precursor for fatty acids. Higher ¹³C incorporation from alanine and glucose into specific PLFAs compared to glutamate, ribose and acetate reflects the preferential use of glycolysis-derived substances in the fatty acids synthesis.Gram-negative bacteria (16:1ω7c and 18:1ω7c) were the most abundant and active in LMWOS utilisation. Their high activity corresponds to a high demand for anabolic products, e.g. to dominance of pentose-phosphate pathway, i.e. incorporation of ribose-C into PLFAs. The ¹³C incorporation from sugars and amino acids into filamentous microorganisms was lower than into all prokaryotic groups. However, for carboxylic acids, the incorporation was in the same range (0.1–0.2% of the applied carboxylic acid ¹³C) as that of gram-positive bacteria. This may reflect the dominance of fungi and other filamentous microorganisms for utilisation of acidic and complex organics.Thus, we showed that despite similar initial uptake, C from individual LMWOS follows deviating metabolic pathways which accounts for the individual fate of LMWOS-C over 10 days. Consequently, stabilisation of C in soil is mainly connected with its incorporation into microbial compounds of various stability and not with its initial microbial uptake.     

Microbial uptake of low‐molecular‐weight organic substances out‐competes sorption in soil

Low‐molecular‐weight organic substances (LMWOS) such as amino acids, sugars and carboxylates, are rapidly turned over in soil. Despite their importance, it remains unknown how the competition between microbial uptake and sorption to the soil matrix affects the LMWOS turnover in soil solution. This study describes the dynamics of LMWOS fluxes (10 µm ) in various pools (dissolved, sorbed, decomposed to CO₂ and incorporated into microbial biomass) and also assesses the LMWOS distribution in these pools over a very wide concentration range (0.01–1000 µm ). Representatives of each LMWOS group (glucose for sugars, alanine for amino acids, acetate for carboxylates), uniformly ¹⁴C‐labelled, were added to sterilized or non‐sterilized soil and analysed in different pools between 1 minute and 5.6 hours after addition. LMWOS were almost completely taken up by microorganisms within the first 30 minutes. Surprisingly, microbial uptake was much faster than the physicochemical sorption (estimated in sterilized soil), which needed 60 minutes to reach quasi‐equilibrium for alanine and about 400 minutes for glucose. Only acetate sorption was instantaneous. At a concentration of 100 µm , microbial decomposition after 4.5 hours was greater for alanine (76.7 ± 1.1%) than for acetate (55.2 ± 0.9%) or glucose (28.5 ± 1.5%). In contrast, incorporation into microbial biomass was greater for glucose (59.8 ± 1.2%) than for acetate (23.4 ± 5.9%) or alanine (5.2 ± 2.8%). Between 10 and 500 µm , the pathways of the three LMWOS changed: at 500 µm , alanine and acetate were less mineralized and more was incorporated into microbial biomass than at 10 µm , while glucose incorporation decreased. Despite the fact that the LMWOS concentrations in soil solution were important for competition between sorption and microbial uptake, their fate in soil is mainly determined by microbial uptake and further microbial transformations. For these substances, which represent the three main groups of LMWOS in soil, the microbial uptake out‐competes sorption.     

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.     

Repeated CO2 pulse-labelling reveals an additional net gain of soil carbon during growth of spring wheat under free air carbon dioxide enrichment (FACE)

Rising levels of atmospheric CO₂ have often been found to increase above and belowground biomass production of C3 plants. The additional translocation of organic matter into soils by increased root mass and exudates are supposed to possibly increase C pools in terrestrial ecosystems. Corresponding investigations were mostly conducted under more or less artificial indoor conditions with disturbed soils. To overcome these limitations, we conducted a ¹⁴CO₂ pulse-labelling experiment within the German FACE project to elucidate the role of an arable crop system in carbon sequestration under elevated CO₂. We cultivated spring wheat cv. “Minaret” with usual fertilisation and ample water supply in stainless steel cylinders forced into the soil of a control and a FACE plot. Between stem elongation and beginning of ripening the plants were repeatedly pulse-labelled with ¹⁴CO₂ in the field. Soil born total CO₂ and ¹⁴CO₂ was monitored daily till harvest. Thereafter, the distribution of ¹⁴C was analysed in all plant parts, soil, soil mineral fractions and soil microbial biomass. Due to the small number of grown wheat plants (40) in each ring and the inherent low statistical power, no significant above and belowground growth effect of elevated CO₂ was detected at harvest. But in comparison to ambient conditions, 28% more ¹⁴CO₂ and 12% more total CO₂ was evolved from soil under elevated CO₂ (550 μmol CO₂ mol⁻¹). In the root-free soil 27% more residual ¹⁴C was found in the FACE soil than in the soil from the ambient ring. In soil samples from both treatments about 80% of residual ¹⁴C was found in the clay fraction and 7% in the silt fraction. Very low ¹⁴C contents in the CFE extracts of microbial biomass in the soil from both CO₂ treatments did not allow assessing their influence on this parameter. Since the calculated specific radioactivity of soil born ¹⁴CO₂ gave no indication of an accelerated priming effect in the FACE soil, we conclude that wheat plants grown under elevated CO₂ can contribute to an additional net carbon gain in soils.     

Microbial immobilisation of C rhizodeposits in rhizosphere and root-free soil under continuous C labelling of oats

A greenhouse experiment was conducted by growing oats (Avenasativa L.) in a continuously ¹³CO₂ labeled atmosphere. The allocation of ¹³C-labeled photosynthates in plants, microbial biomass in rhizosphere and root-free soil, pools of soil organic C, and CO₂ emissions were examined over the plant's life cycle. To isolate rhizosphere from root-free soil, plant seedlings were placed into bags made of nylon monofilament screen tissue (16 μm mesh) filled with soil. Two peaks of ¹³C in rhizosphere pools of microbial biomass and dissolved organic carbon (DOC), as well as in CO₂ emissions at the earing and ripeness stages were revealed. These ¹³C maxima corresponded to: (i) the end of rapid root growth and (ii) beginning of root decomposition, respectively. The δ¹³C values of microbial biomass were higher than those of DOC and of soil organic matter (SOM). The microbial biomass C accounted for up to 56 and 39% of ¹³C recovered in the rhizosphere and root-free soil, respectively. Between 4 and 28% of ¹³C assimilated was recovered in the root-free soil. Depending on the phenological stage, the contribution of root-derived C to total CO₂ emission from soil varied from 61 to 92% of total CO₂ evolved, including 4-23% attributed to rhizomicrobial respiration. While 81-91% of C substrates used for microbial growth in the root-free soil and rhizosphere came from SOM, the remaining 9-19% of C substrates utilized by the microbial biomass was attributable to rhizodeposition. The use of continuous isotopic labelling and physical separation of root-free and rhizosphere soil, combined with natural ¹³C abundance were effective in gaining new insight on soil and rhizosphere C-cycling.     

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.     

Decomposition of N-labelled maize leaves in soil affected by endogeic geophagous Aporrectodea caliginosa

A microcosm experiment was carried out for 56 days at 12 °C to evaluate the feeding effects of the endogeic geophagous earthworm species Aporrectodea caliginosa on the microbial use of ¹⁵N-labelled maize leaves (Zea mays) added as 5 mm particles equivalent to 1 mg C and 57 μg N g⁻¹ soil. The dry weight of A. caliginosa biomass decreased in the no-maize treatment by 10% during the incubation and increased in the maize leaf treatments by 18%. Roughly 5% and 10% of the added maize leaf-C and leaf-N, respectively, were incorporated into the biomass of A. caliginosa. About 29% and 33% of the added maize leaf-C were mineralised to CO₂ in the no-earthworm and earthworm treatments, respectively. The presence of A. caliginosa significantly increased soil-derived CO₂ production by 90 μg g⁻¹ soil in the no-maize and maize leaf treatments, but increased the maize-derived CO₂ production only by 40 μg g⁻¹ soil. About 10.5% of maize leaf-C and leaf-N was incorporated into the soil microbial biomass in the absence of earthworms, but only 6% of the maize leaf-C and 3% of the maize leaf-N in the presence of earthworms. A. caliginosa preferentially fed on N rich, maize leaf-colonizing microorganisms to meet its N demand. This led to a significantly increased C/N ratio of the unconsumed microbial biomass in soil. The ergosterol-to-microbial biomass C ratio was not significantly decreased by the presence of earthworms. A. caliginosa did not directly contribute to comminution of plant residues, as indicated by the absence of any effects on the contents of the different particulate organic matter fractions, but mainly to grazing of residue-colonizing microorganisms, increasing their turnover considerably.     

Dynamics of labile and recalcitrant soil carbon pools in a sorghum free-air CO2 enrichment (FACE) agroecosystem

Experimentation with dynamics of soil carbon pools as affected by elevated CO₂ can better define the ability of terrestrial ecosystems to sequester global carbon. In the present study, 6 N HCl hydrolysis and stable-carbon isotopic analysis (δ¹³C) were used to investigate labile and recalcitrant soil carbon pools and the translocation among these pools of sorghum residues isotopically labeled in the 1998-1999 Arizona Maricopa free air CO₂ enrichment (FACE) experiment, in which elevated CO₂ (FACE: 560 μmol mol⁻¹) and ambient CO₂ (Control: 360 μmol mol⁻¹) interact with water-adequate (wet) and water-deficient (dry) treatments. We found that on average 53% of the final soil organic carbon (SOC) in the FACE plot was in the recalcitrant carbon pool and 47% in the labile pool, whereas in the Control plot 46% and 54% of carbon were in recalcitrant and labile pools, respectively, indicating that elevated CO₂ transferred more SOC into the slow-decay carbon pool. Also, isotopic mixing models revealed that increased new sorghum residue input to the recalcitrant pool mainly accounts for this change, especially for the upper soil horizon (0-30 cm) where new carbon in recalcitrant soil pools of FACE wet and dry treatments was 1.7 and 2.8 times as large as that in respective Control recalcitrant pools. Similarly, old C in the recalcitrant pool under elevated CO₂ was higher than that under ambient CO₂, indicating that elevated CO₂ reduces the decay of the old C in recalcitrant pool. Mean residence time (MRT) of bulk soil carbon at the depth of 0-30 cm was significantly longer in FACE plot than Control plot by the averages of 12 and 13 yr under the dry and wet conditions, respectively. The MRT was positively correlated to the ratio of carbon content in the recalcitrant pool to total SOC and negatively correlated to the ratio of carbon content in the labile pool to total SOC. Influence of water alone on the bulk SOC or the labile and recalcitrant pools was not significant. However, water stress interacting with CO₂ enhanced the shift of the carbon from labile pool to recalcitrant pool. Our results imply that terrestrial agroecosystems may play a critical role in sequestrating atmospheric CO₂ and mitigating harmful CO₂ under future atmospheric conditions.     

Changes in the fungal-to-bacterial respiratory ratio and microbial biomass in agriculturally managed soils under free-air CO2 enrichment (FACE) - A six-year survey of a field study

In soil ecology, microbial parameters have been identified as sensitive indicators of changes in the soil environment. The Braunschweig FACE project provided the opportunity to study the effects of elevated CO₂ (550 μmol mol⁻¹) as compared to ambient CO₂ (370 μmol mol⁻¹) on total microbial biomass (C_mic), C_mic-to-C_org ratio and the fungal-to-bacterial respiratory ratio together with total C_org, N_t, C:N ratio and pH over a six-year period. Field management followed a typical crop rotation system of this region with either a crop-related full nitrogen supply (N100) or 50% reduced N supply (N50). The soil microbial parameters responded to the elevated CO₂ treatment in varying intensities and time spans. The fungal-to-bacterial respiratory ratio was the most sensitive parameter in responding to an elevated CO₂ treatment with highly significant differences to ambient CO₂-treated control plots in the third year of CO₂ fumigation. After six years bacterial respiratory activity had increased in ascending order to 34% in FACE-treated plots (N50 and N100) as compared to control plots. Soil microbial biomass (C_mic) responded more slowly to the FACE treatment with highly significant increases of >12% after the fourth year of CO₂ fumigation. The C_mic-to-C_org ratio responded very late in the last two years of the CO₂ treatment with a significant increase of >7.0% only in the N100 variant. Total C_org and N_t were slightly but significantly increased under FACE around 10.0% with ascending tendency over time starting with the second year of CO₂ treatment. No significant FACE effects could be recorded for the C:N ratio or pH.These results suggest that under FACE treatment changes in the soil microbial community will occur. In our study the fungal-to-bacterial respiratory ratio was superior to total C_mic as microbial bioindicators in reflecting changes in the soil organic matter composition.     

Rhizodeposition-induced decomposition increases N availability to wild and cultivated wheat genotypes under elevated CO2

Elevated CO₂ may increase nutrient availability in the rhizosphere by stimulating N release from recalcitrant soil organic matter (SOM) pools through enhanced rhizodeposition. We aimed to elucidate how CO₂-induced increases in rhizodeposition affect N release from recalcitrant SOM, and how wild versus cultivated genotypes of wheat mediated differential responses in soil N cycling under elevated CO₂. To quantify root-derived soil carbon (C) input and release of N from stable SOM pools, plants were grown for 1 month in microcosms, exposed to ¹³C labeling at ambient (392 μmol mol⁻¹) and elevated (792 μmol mol⁻¹) CO₂ concentrations, in soil containing ¹⁵N predominantly incorporated into recalcitrant SOM pools. Decomposition of stable soil C increased by 43%, root-derived soil C increased by 59%, and microbial-¹³C was enhanced by 50% under elevated compared to ambient CO₂. Concurrently, plant ¹⁵N uptake increased (+7%) under elevated CO₂ while ¹⁵N contents in the microbial biomass and mineral N pool decreased. Wild genotypes allocated more C to their roots, while cultivated genotypes allocated more C to their shoots under ambient and elevated CO₂. This led to increased stable C decomposition, but not to increased N acquisition for the wild genotypes. Data suggest that increased rhizodeposition under elevated CO₂ can stimulate mineralization of N from recalcitrant SOM pools and that contrasting C allocation patterns cannot fully explain plant mediated differential responses in soil N cycling to elevated CO₂.     

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

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

Short and long-term effects of elevated CO2 on Lolium perenne rhizodeposition and its consequences on soil organic matter turnover and plant N yield

It is still unclear whether elevated CO₂ increases plant root exudation and consequently affects the soil microbial biomass. The effects of elevated CO₂ on the fate of the C and nitrogen (N) contained in old soil organic matter pools is also unclear. In this study the short and long-term effects of elevated CO₂ on C and N pools and fluxes were assessed by growing isolated plants of ryegrass (Lolium perenne) in glasshouses at elevated and ambient atmospheric CO₂ and using soil from the New Zealand FACE site that had >4 years exposure to CO₂ enrichment. Using ¹⁴CO₂ pulse labelling, the effects of elevated CO₂ on C allocation within the plant-soil system were studied. Under elevated CO₂ more root derived C was found in the soil and in the microbial biomass 48 h after labelling. The increased availability of substrate significantly stimulated soil microbial growth and acted as priming effect, enhancing native soil organic matter decomposition regardless of the mineral N supply. Despite indications of faster N cycling in soil under elevated CO₂, N availability to plants stayed unchanged. Soil previously exposed to elevated CO₂ exhibited a higher N cycling rate but again there was no effect on plant N uptake. With respect to the difficulties of extrapolating glasshouse experiment results to the field, we concluded that the accumulation of coarse organic matter observed in the field under elevated CO₂ was probably not created by an imbalance between C and N but was likely to be due to more complex phenomena involving soil mesofauna and/or other nutrients limitations.     

Microbial use of maize cellulose and sugarcane sucrose monitored by changes in the C/C ratio

An arable soil with organic matter formed from C₃-vegetation was amended initially with maize cellulose (C₄-cellulose) and sugarcane sucrose (C₄-sucrose) in a 67-day laboratory incubation experiment with microcosms at 25 °C. The amount and isotopic composition (¹³C/¹²C) of soil organic C, CO₂ evolved, microbial biomass C, and microbial residue C were determined to prove whether the formation of microbial residues depends on the quality of the added C source adjusted with NH₄NO₃ to the same C/N ratio of 15. In a subsequent step, C₃-cellulose (3 mg C g⁻¹ soil) was added without N to soil to determine whether the microbial residues formed initially from C₄-substrate are preferentially decomposed to maintain the N-demand of the soil microbial community. At the end of the experiment, 23% of the two C₄-substrates added was left in the soil, while 3% and 4% of the added C₄-cellulose and C₄-sucrose, respectively, were found in the microbial biomass. The addition of the two C₄-substrates caused a significant 100% increase in C₃-derived CO₂ evolution during the 5-33 day incubation period. The addition of C₃-cellulose caused a significant 50% increase in C₄-derived CO₂ evolution during the 38-67 day incubation period. The decrease in microbial biomass C₄-C accounted for roughly 60% of this increase. Cellulose addition promoted microorganisms strongly able to recycle N immediately from their own tissue by “cryptic growth” instead of incorporating NO₃⁻ from the soil solution. The differences in quality of the microbial residues produced by C₄-cellulose and C₄-sucrose decomposing microorganisms are also reflected by the difference in the rates of CO₂ evolution, but not in the rates of net N mineralization.     

CO2 and inorganic N supply modify competition for N between co-occurring grass plants, tree seedlings and soil microorganisms

Plant-plant and plant-soil interactions play a key role in determining plant community structure and ecosystem function. However, the effects of global change on the interplay between co-occurring plants and soil microbes in successional communities are poorly understood. In this study, we investigated competition for nitrogen (N) between soil microorganisms, grass plants and establishing tree seedlings under factorial carbon dioxide (CO₂) and N treatments. Fraxinus excelsior seedlings were germinated in the presence or absence of grass competition (Dactylis glomerata) at low (380 μmol mol⁻¹) or high (645 μmol mol⁻¹) CO₂ and at two levels of N nutrition in a mesocosm experiment. Pulse ¹⁵N labelling was used to examine N partitioning among plant and soil compartments. Dactylis exerted a strong negative effect on Fraxinus biomass, N capture and ¹⁵N recovery irrespective of N and CO₂ treatment. In contrast, the presence of Dactylis had a positive effect on the microbial N pool. Plant and soil responses to N treatment were of a greater magnitude compared with responses to elevated CO₂, but the pattern of Fraxinus- and microbial-N pool response to N and CO₂ varied depending on grass competition treatment. Within the Dactylis competition treatment, decreases in Fraxinus biomass in response to N were not mirrored by decreases in tree seedling N content, suggesting a shift from below- to above-ground competition. In the Dactylis-sown pots, ¹⁵N recovery could be ranked Dactylis > microbial pool > Fraxinus in all N and CO₂ treatment combinations. Inequalities between Fraxinus and soil microorganisms in terms of ¹⁵N recovery were exacerbated by N addition. Contrary to expectations, elevated CO₂ did not increase plant-microbe competition. Nevertheless, microbial ¹⁵N recovery showed a small positive increase in the high CO₂ treatment. Overall, elevated CO₂ and N supply did not interact on plant/soil N partitioning. Our data suggest that the competitive balance between establishing tree seedlings and grass plants in an undisturbed sward is relatively insensitive to CO₂ or N-induced modifications in N competition between plant and soil compartments.     

Short-term decomposition of <Superscript>14</Superscript>C-labelled glucose in a fluvo-aquic soil as affected by lanthanum amendment

In this study, we investigated the effects of lanthanum (La), one of the rare earth elements (REEs), on microbial biomass C as well as the decomposition of ¹⁴C-labelled glucose in a fluvo-aquic soil in 28 days. The soil was collected from the field plots under maize/wheat rotation in Fengqiu Ecological Experimental Station of Chinese Academy of Sciences, Henan Province, China. Application of La decreased soil microbial biomass C during the experimental period, and there was a negative correlation (P < 0.01) between microbial biomass and application rate of La. La increased microbial biomass ¹⁴C after ¹⁴C glucose addition, and the increase was significant (P < 0.05) at the rates of more than 160 mg kg⁻¹ soil. La slightly increased ¹⁴CO₂ evolution at lower rates of application but decreased it at higher rates 1 day after ¹⁴C glucose addition, while there was no significant effect from days 2 to 28. For the cumulative ¹⁴CO₂ evolution during the incubation of 28 days, La slightly increased it at the rates of less than 120 mg kg⁻¹ soil, while significantly decreased (P < 0.05) it at the rate of 200 mg kg⁻¹ soil. The results indicated that agricultural use of REEs such as La in soil could decrease the amount of soil microbial biomass and change the pattern of microbial utilization on glucose C source in a short period.     

Increased microbial activity under elevated [CO2] does not enhance residue decomposition in a semi-arid cropping system in Australia

The association between the responses of microbial activity and residue decomposition to elevated atmospheric [CO₂] under field conditions in Australian cropping systems is unknown. We measured soil CO₂ emission and decomposition of wheat and field pea residues in a wheat cropping system in the field using the Australian Grains Free-Air CO₂ Enrichment (AGFACE) facility in Horsham, Victoria. Elevated [CO₂] (550 μmol mol⁻¹) increased soil CO₂ emission by 41%, but did not affect the percentage of the original mass or C remaining for either type of residue throughout the experimental period. Our findings suggest that the rates of residue decomposition and residue C mineralization in this semi-arid wheat cropping system were not affected by elevated [CO₂] despite higher microbial activity. This has major implication for the C sequestration potential of semi-arid cropping systems under future CO₂ climates.     

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.     

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.     

Origin of positive δC of emitted CO2 from soils after application of biogas residues

Bioenergy production from renewable organic material is known to be a clean energy source and therefore its use is currently much promoted in many countries. Biogas by-products also called biogas residues (BGR) are rich in partially stable organic carbon and can be used as an organic fertilizer for crop production. However so far, many environmental issues relevant when BGR are applied to agricultural land (soil C sequestration, increased denitrification and nutrient leaching) still have to be studied. Therefore a field experiment was set up to investigate the degradation of BGR and its impact on the decomposition of native soil organic matter based on a natural abundance stable isotope approach. Maize, a C₄ plant has been used as bioenergy crop, therefore the δ¹³C of total C in BGR was −16.0‰_PDB and soil organic matter was mostly derived from C₃ plant based detritus, SOM thus showed a δ¹³C of −28.4‰_PDB. Immediately after BGR application, soil-emitted CO₂ showed unexpectedly high δ¹³C of up to +23.6‰_PDB, which has never been reported earlier. A subsequent laboratory scale experiment confirmed the positive δ¹³C of soil-emitted CO₂ after BGR addition and showed that obviously, the added BGR led to a consumption of dissolved inorganic C in soils. Additionally, it was observed that the δ¹³C of CO₂ driven from inorganic C of BGR (BGR-IC) by acid treatment was +35.6‰_PDB. Therefore, we suggest that also under field conditions the transformation of BGR-IC into CO₂ contributed largely to CO₂ emissions in addition to the decomposition of organic matter, which affected both the amount and the carbon isotope signature of emitted CO₂ in the initial period after BGR application. Positive δ¹³C of inorganic C contained in BGR was attributed to processes with strong fractionation of C isotopes during anaerobic fermentation in the biogas formation process.