Dynamics of maize (Zea mays L.) leaf straw mineralization as affected by the presence of soil and the availability of nitrogen

An incubation experiment was carried out with maize (Zea mays L.) leaf straw to analyze the effects of mixing the residues with soil and N amendment on the decomposition process. In order to distinguish between soil effects and nitrogen effects for both the phyllospheric microorganisms already present on the surface of maize straw and soil microorganisms the N amendment was applied in two different placements: directly to the straw or to the soil. The experiment was performed in dynamic, automated microcosms for 22 days at 15 °C with 7 treatments: (1) untreated soil, (2) non-amended maize leaf straw without soil, (3) N amended maize leaf straw without soil, (4) soil mixed with maize leaf straw, (5) N amended soil, (6) N amended soil mixed with maize leaf straw, and (7) soil mixed with N amended maize leaf straw. ¹⁵NH₄¹⁵NO₃ (5 at%) was added. Gas emissions (CO₂, ¹³CO₂ and N₂O) were continuously recorded throughout the experiment. Microbial biomass C, biomass N, ergosterol, δ¹³C of soil organic C and of microbial biomass C as well as ¹⁵N in soil total N, mineral N and microbial biomass N were determined in soil samples at the end of the incubation. The CO₂ evolution rate showed a lag-phase of two days in the non-amended maize leaf straw treatment without soil, which was completely eliminated when mineral N was added. The addition of N generally increased the CO₂ evolution rate during the initial stages of maize leaf straw decomposition, but not the cumulative CO₂ production. The presence of soil caused roughly a 50% increase in cumulative CO₂ production within 22 days in the maize straw treatments due to a slower decrease of CO₂ evolution after the initial activity peak. Since there are no limitations of water or N, we suggest that soil provides a microbial community ensuring an effective succession of straw decomposing microorganisms. In the treatments where maize and soil was mixed, 75% of microbial biomass C was derived from maize. We concluded that this high contribution of maize using microbiota indicates a strong influence of organisms of phyllospheric origin to the microbial community in the soil after plant residues enter the soil.     

Formation and use of microbial residues after adding sugarcane sucrose to a heated soil devoid of soil organic matter

A 67-day incubation experiment was carried out with a soil initially devoid of any organic matter due to heating, which was amended with sugarcane sucrose (C₄-sucrose with a δ¹³C value of ?10.5‰), inorganic N and an inoculum for recolonisation and subsequently at day 33 with C₃-cellulose (δ¹³C value of ?23.4‰). In this soil, all organic matter is in the microbial biomass or in freshly formed residues, which makes it possible to analyse more clearly the role of microbial residues for decomposition of N-poor substrates. The average δ¹³C value over the whole incubation period was ?10.7‰ in soil total C in the treatments without C₃-cellulose addition. In the CO₂ evolved, the δ¹³C values decreased from ?13.4‰ to ?15.4‰ during incubation. In the microbial biomass, the δ¹³C values increased from ?11.5‰ to ?10.1‰ at days 33 and 38. At day 67, 36% of the C₄-sucrose was left in the treatment without a second amendment. The addition of C₃-cellulose resulted in a further 7% decrease, but 4% of the C₃-cellulose was lost during the second incubation period. Total microbial biomass C declined from 200 μg g^?1 soil at day 5 to 70 μg g^?1 soil at day 67. Fungal ergosterol increased to 1.5 μg g^?1 soil at day 12 and declined more or less linearly to 0.4 μg g^?1 soil at day 67. Bacterial muramic acid declined from a maximum of 35 μg g^?1 soil at day 5 to a constant level of around 16 μg g^?1 soil. Glucosamine showed a peak value at day 12. Galactosamine remained constant throughout the incubation. The fungal C/bacterial C ratio increased more or less linearly from 0.38 at day 5 to 1.1 at day 67 indicating a shift in the microbial community from bacteria to fungi during the incubation. The addition of C₃-cellulose led to a small increase in C₃-derived microbial biomass C, but to a strong increase in C₄-derived microbial biomass C. At days 45 and 67, the addition of N-free C₃-cellulose significantly decreased the C/N ratio of the microbial residues, suggesting that this fraction did not serve as an N-source, but as an energy source.     

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.     

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

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

Three-source partitioning of 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.     

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.     

C and N turnover in structurally intact soils of different texture

The turnover of native and applied C and N in undisturbed soil samples of different texture but similar mineralogical composition, origin and cropping history was evaluated at −10 kPa water potential. Cores of structurally intact soil with 108, 224 and 337 g clay kg⁻¹ were horizontially sliced and ¹⁵N-labelled sheep faeces was placed between the two halves of the intact core. The cores together with unamended treatments were incubated in the dark at 20 °C and the evolution of CO₂-C determined continuously for 177 d. Inorganic and microbial biomass N and ¹⁵N were determined periodically. Net nitrification was less in soil amended with faeces compared with unamended soil. When adjusted for the NO₃-N present in soil before faeces was applied, net nitrification became negative indicating that NO₃-N had been immobilized or denitrified. The soil most rich in clay nitrified least N and ¹⁵N. The amounts of N retained in the microbial biomass in unamended soils increased with clay content. A maximum of 13% of the faeces ¹⁵N was recovered in the microbial biomass in the amended soils. CO₂-C evolution increased with clay content in amended and unamended soils. CO₂-C evolution from the most sandy soil was reduced due to a low content of potentially mineralizable native soil C whereas the rate constant of C mineralization rate peaked in this soil. When the pool of potentially mineralizable native soil C was assumed proportional to volumetric water content, the three soils contained similar proportions of potentially mineralizable native soil C but the rate constant of C mineralization remained highest in the soil with least clay. Thus although a similar availability of water in the three soils was ensured by their identical matric potential, the actual volume of water seemed to determine the proportion of total C that was potentially mineralizable. The proportion of mineralizable C in the faeces was similar in the three soils (70% of total C), again with a higher rate constant of C mineralization in the soil with least clay. It is hypothesized that the pool of potentially mineralizable C and C rate constants fluctuate with the soil water content.     

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.     

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.     

Immobilization and mineralization of nitrogen in a saline and alkaline soil during microbial use of sugarcane filter cake amended with glucose

A 42-day incubation was conducted to study the effect of glucose and ammonium addition adjusted to a C/N ratio of 12.5 on sugarcane filter cake decomposition and on the release of inorganic N from microbial residues formed initially. The CO₂ evolved increased in comparison with the non-amended control from 35% of the added C with pure +5 mg g⁻¹ soil filter cake amendment to 41% with +5 mg g⁻¹ soil filter cake +2.5 mg g⁻¹ soil glucose amendment to 48% with 5 mg g⁻¹ soil filter cake +5 mg g⁻¹ soil glucose amendment. The different amendments increased microbial biomass C and microbial biomass N within 6 h and such an increase persisted. The fungal cell-membrane component ergosterol initially showed a disproportionate increase in relation to microbial biomass C, which completely disappeared by the end of the incubation. The cellulase activity showed a 5-fold increase after filter cake addition, which was not further increased by the additional glucose amendment. The cellulase activity showed an exponential decline to values around 4% of the initial value in all treatments. The amount of inorganic N immobilized from day 0 to day 14 increased with increasing amount of C added, in contrast to the control treatment. After day 14, the immobilized N was re-mineralized at rates between 1.3 and 1.5 μg N g⁻¹ soil d⁻¹ in the treatments being more than twice as high as in the control treatment. This means that the re-mineralization rate is independent of the actual size of the microbial residues pool and also independent of the size of the soil microbial biomass.     

Interactions of NaCl and Na2SO4 on soil organic C mineralization after addition of maize straws

NaCl and Na₂SO₄ often dominate salt compositions in saline soils. While either salt alone affects soil organic matter mineralization, their interactions on soil organic matter dynamics are unknown. This study aimed to investigate interactive effects of the two salts on organic C mineralization and microbial biomass C of the saline soils after addition of maize straws. Both NaCl and Na₂SO₄ were applied at 0, 40 and 80 mmol Na kg⁻¹ soil and the incubation was undertaken at soil water content of 15% and 20% (w/w) in dark at 28.5 °C for 70 days. The study found significant interactions of NaCl and Na₂SO₄ on CO₂-C evolution during the early incubation periods—a suppressing effect at days 1-2 but a stimulating effect at days 6-8 and 17-20, and thereafter the salt interactions were influenced by water content. The interactions of water content with NaCl or Na₂SO₄ on CO₂-C evolution were observed through the incubation periods except days 1-2, showing that the salt effects were dependent on water content. Total CO₂ evolution over the 70-day-long incubation decreased with increasing NaCl but increased with increasing Na₂SO₄ compared to the nil-salted treatment. Salt interactions on soil microbial biomass C were observed at days 7, 21, but not at day 49. Microbial biomass C increased at day 7 in the soils treated with either NaCl or Na₂SO₄ but decreased where the two salts were combined. At day 21, microbial biomass C increased with NaCl but decreased with Na₂SO₄ regardless whether the counterpart salt was added. The results suggest that soil organic C mineralization can be affected by the interactions of NaCl and Na₂SO₄, possibly through the salt-induced changes in microbial biomass community structure.     

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.     

Microbial immobilisation and turnover of C labelled substrates in two arable soils under field and laboratory conditions

Microbial biomass C immobilisation and turnover were studied under field and laboratory conditions in soils of high yield (HY) and low yield (LY) areas within an agricultural field. We compared the size and activity of soil microbial biomass (SMB) in the soils of the different yield areas under field and laboratory conditions. Soils were amended with ¹³C labelled mustard (Sinapis alba) residues (both experiments) and labelled glucose (laboratory only) at 500 μg C g⁻¹ dry soil. SMB-C, dissolved organic carbon (DOC) and total C content were monitored in the field and the laboratory. CO₂-efflux was also measured in laboratory treatments. Isotope ratios were determined for SMB in both experiments, but other variables only in the laboratory treatments. A positive priming effect was measured in three of four laboratory treatments. Priming was induced after a significant increase of soil derived C in the microbial biomass. Thereafter, the total C loss through priming was always smaller than or equal to the decline in microbial biomass C. In field and laboratory experiments SMB in the HY soil immobilised less of the added substrate C than LY soil SMB. Calculated turnover times in the laboratory glucose amendment were 0.24 (HY) and 0.31 y (LY), in the laboratory mustard treatment 0.58 (HY) and 0.44 y (LY) and in the field mustard amendments 1.09 (HY) and 1.25 y (LY). In both the field mustard and laboratory glucose treatments turnover in the HY soil tended to exceed that in the LY soil. These turnover times as well as the reaction of SMB-C to drying-rewetting and substrate addition, indicated that the HY soil possessed a more active microbial community with a more rapid C turnover than the LY soil. As C turnover is considered to be closely linked to nutrient cycles, faster turnover in the HY soil may involve a better nutrient supply for crops resulting in higher agricultural yield.     

Adenylates in the soil microbial biomass at different temperatures

Five soils from temperate sites (Germany; 2 arable and 3 grassland) were incubated aerobically at 5, 10, 15, 20, 25, 35, and 40 °C for 8 days. Soils were analysed for soil microbial biomass C, biomass N, AMP, ADP, and ATP to determine whether the increase in the ATP-to-microbial biomass C ratio with increasing temperature was either due to an increase in the adenylate energy charge (AEC) or de novo synthesis of ATP, or both. Around 80% of the variance in microbial biomass C and biomass N was explained by differences in soil properties, only 7% by the temperature treatments. Averaging the data of all 5 soils for each incubation temperature, the microbial biomass C content decreased with increasing temperature from 15 to 40 °C continuously by 2.5 μg g⁻¹ soil °C⁻¹ after 8-days' incubation. However, this decrease was not accompanied by a similar decrease in microbial biomass N. The average microbial biomass C/N ratio was 6.8. Between 54 and 76% of the variance in AMP, ADP, ATP and the sum of adenylates was explained by differences in soil properties and between 14 (ADP) and 27% (ATP) by the temperature treatments. However, temperature effects on AMP and ADP were variable and inconsistent. In contrast, ATP and consequently also the sum of adenylates increased continuously from 5 to 30 °C followed by a decline to 40 °C. The AEC showed similarly a small, but significant increase with increasing temperature from 0.73 to 0.85 at 30 °C. Consequently, the majority of the variance, i.e. roughly 60% in AEC values, but also in ATP-to-microbial biomass C ratios was explained by the incubation temperature. The mean ATP-to-microbial biomass C ratio increased from 4.7 μmol g⁻¹ at 5 °C to a 2.5 fold maximum of 12.0 μmol g⁻¹ at 35 °C. This increase was linear with a rate of 0.26 μmol ATP g⁻¹ microbial biomass C °C⁻¹. The energy for the extra ATP produced during temperature increase is probably derived from an accelerated turnover of endocellular C reserves in the microbial biomass.     

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.     

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

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

The proportional mineralisation of microbial biomass and organic matter caused by air-drying and rewetting of a grassland soil

During the first few days after rewetting of an air-dried soil (AD-RW), microbial activity increases compared to that in the original moist soil, causing increased mineralisation (a flush) of soil organic carbon (C) and other nutrients. The AD-RW flush is believed to be derived from the enhanced mineralisation of both non-biomass soil organic matter (due to its physical release and enhanced availability) and microbial biomass killed during drying and rewetting. Our aim was to determine the effects of AD-RW on the mineralisation of soil organic matter and microbial biomass during and after repeated AD-RW cycles and to quantify their proportions in the CO₂-C flushes that resulted. To do this, a UK grassland soil was amended with ¹⁴C-labelled glucose to label the biomass and then given five AD-RW cycles, each followed by 7 d incubation at 25 °C and 50% water holding capacity. Each AD-RW cycle increased the amount of CO₂-C evolved (varying from 83 to 240 μg g⁻¹ soil), compared to the control with, overall, less CO₂-C being evolved as the number of AD-RW cycles increased. In the first cycle, the amount of biomass C decreased by 44% and microbial ATP by 70% while concentrations of extractable C nearly doubled. However, all rapidly recovered and within 1.3 d after rewetting, biomass C was 87% and ATP was 78% of the initial concentrations measured prior to air-drying. Similarly, by 2 d, extractable organic C had decreased to a similar concentration to the original. After the five AD-RW cycles, the amounts of total and ¹⁴C-labelled biomass C remaining in the soil accounted for 60 and 40% of those in the similarly incubated control soil, respectively. Soil biomass ATP concentrations following the first AD-RW cycle remained remarkably constant (ranging from about 10 to 14 μmol ATP g⁻¹ biomass C) and very similar to the concentration in the fresh soil prior to air-drying. We developed a simple mathematical procedure to estimate the proportion of CO₂-C derived from biomass C and non-biomass C during AD-RW. From it, we estimate that, over the five AD-RW cycles, about 60% of the CO₂-C evolved came from mineralisation of non-biomass organic C and the remainder from the biomass C itself.     

Fate of gram-negative bacterial biomass in soil—mineralization and contribution to SOM

Soil microorganisms contribute to the formation of non-living soil organic matter (SOM) by metabolic transformation of plant-derived material. After cell death, their biomass components with a specific molecular character become incorporated into SOM imprinting its chemical properties, although this process has not yet been quantified. In order to elucidate the contribution to SOM formation, we investigated the fate of gram-negative bacterial model biomass (Escherichia coli usually introduced into soil with manure or feces) during incubation of soil with isotopically (¹³C) and genetically (lux gene) labeled cells. The decline of living cells was monitored by the loss of bioluminescence. The carbon turnover and mineralization was balanced by bulk soil stable isotope analysis, and the persistence of nucleic acids was investigated by PCR amplification of the lux gene. During incubation, the number of viable E. coli cells decreased rapidly (99.9% within the first 42 d) serving as substrate for other microorganisms or for the formation of SOM, and bioluminescent cells could only be detected during the first 56 d. However, the lux gene was still detected after 224 d, which indicates stabilization of DNA in SOM. Although the survival of E. coli in soil is limited, only about 65% of the added labeled biomass carbon was mineralized to ¹³CO₂ and 51% remained in soil after 224 d with an average ¹³C recovery of 117%. The amount of ¹³C found in the PLFA representative of living cells had decreased to 25% of the initial value, suggesting a proportional decrease of the ¹³C in the soil microbial biomass. The extent of this decrease is higher than the mineralization of the bulk E. coli C and thus the difference of around 25% has to be stabilized as metabolites, or in non-living SOM. The data provide evidence that the genetic information and a considerable part of the carbon from dying bacterial biomass were retained in both the soil microbial food web and in non-living SOM.     

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

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

Respiration pattern and microbial use of field-grown transgenic Bt-maize residues

A 49-day incubation experiment was carried out with the addition of field-grown maize stem and leaf residues to soil at three different temperatures (5, 15, and 25 °C). The aim was to study the effects of two transgenic Bt-maize varieties in comparison to their two parental non-Bt varieties on the mineralization of the residues, on their incorporation into the microbial biomass and on changes in the microbial community structure. The stem and leaf residues of Novelis-Bt contained 3.9 μg g⁻¹ dry weight of the Bt toxin Cry1Ab and those of Valmont-Bt only 0.8 μg g⁻¹. The residues of the two parental non-Bt varieties Nobilis and Prelude contained higher concentrations of ergosterol (+220%) and glucosamine (+190%) and had a larger fungal C-to-bacterial C ratio (+240%) than the two Bt varieties. After adding the Bt residues, an initial peak in respiration of an extra 700 μg CO₂-C g⁻¹ soil or 4% of the added amount was observed in comparison to the two non-Bt varieties at all three temperatures. On average of the four varieties, 19-38% of the maize C added was mineralized during the 49-day incubation at the three different temperatures. The overall mean increase in total maize-derived CO₂ evolution corresponded to a Q₁₀ value of 1.4 for both temperature steps, i.e. from 5 to 15 °C and from 15 to 25 °C. The addition of maize residues led to a strong increase in all microbial properties analyzed. The highest contents were always measured at 5 °C and the lowest at 25 °C. The variety-specific contents of microbial biomass C, biomass N, ATP and adenylates increased in the order Novelis-Bt ? Prelude<Valmont-Bt ? Nobilis. The mineralization of Novelis-Bt residues with the highest Bt concentration and lowest N concentration and their incorporation into the microbial biomass was significantly reduced compared to the parental non-Bt variety Nobilis. These negative effects increased considerably from 5 to 25 °C. The transgenic Bt variety Valmont did not show further significant effects except for the initial peak in respiration at any temperature.