Long-term organic phosphorus mineralization in Spodosols under forests and its relation to carbon and nitrogen mineralization

In forest soils where a large fraction of total phosphorus (P) is in organic forms, soil micro-organisms play a major role in the P cycle and plant availability since they mediate organic P transformations. However, the correct assessment of organic P mineralization is usually a challenging task because mineralized P is rapidly sorbed and most mineralization fluxes are very weak. The objectives of the present work were to quantify in five forest Spodosols at soil depths of 0-15 cm net mineralization of total organic P and the resulting increase in plant available inorganic P and to verify whether net or gross P mineralization could be estimated using the C or N mineralization rates. Net mineralization of total organic P was derived from the net changes in microbial P and gross mineralization of P in dead soil organic matter. We studied very low P-sorbing soils enabling us to use lower extractants to assess the change in total inorganic P as a result of gross mineralization of P in dead soil organic matter. In addition, to enable detection of gross mineralization of P in dead soil organic matter, a long-term incubation (517 days) experiment was carried out. At the beginning of the experiment, total P contents of the soils were very low (19-51 μg g⁻¹) and were essentially present as organic P (17-44 μg g⁻¹, 85-91%) or microbial P (6-14 μg g⁻¹; 24-39%). Conversely, the initial contents of inorganic P were low (2-7 μg g⁻¹; 9-15%). The net changes in the pool size of microbial P during the 517 days of incubation (4-8 μg g⁻¹) and the amounts of P resulting from gross mineralization of dead soil organic matter (0.001-0.018 μg g⁻¹ day⁻¹; 0.4-9.5 μg g⁻¹ for the entire incubation period) were considerable compared to the initial amounts of organic P and also when compared to the initial diffusive iP fraction (<0.3 μg g⁻¹). Diffusive iP corresponds to the phosphate ions that can be transferred from the solid constituents to the soil solution under a gradient of concentration. Net mineralization of organic P induced an important increase in iP in soil solution (0.6-10 μg g⁻¹; 600-5000% increase) and lower increases in diffusive iP fractions (0.3-5 μg g⁻¹; 300-2000% increase), soil solid constituents having an extremely low reactivity relative to iP. Therefore, soil micro-organisms and organic P transformations play a major role in the bioavailability of P in these forest soils. In our study, the dead soil organic matter was defined as a recalcitrant organic fraction. Probably because gross mineralization of P from this recalcitrant organic fraction was mainly driven by the micro-organisms’ needs for energy, the rates of gross mineralization of C, N and P in the recalcitrant organic fraction were similar. Indirect estimation of gross mineralization of P in dead soil organic matter using the gross C mineralization rate seems thus an alternative method for the studied soils. However, additional studies are needed to verify this alternative method in other soils. No relationships were found between microbial P release and microbial C and N releases.     

A mass-balance approach to measuring microbial uptake and pools of phosphorus in nutrient-amended soils

Soil microbial biomass P is usually determined through fumigation-extraction (FE), in which partially extractable P from lysed biomass is converted to biomass P using a conversion factor (K_p). Estimation of K_p has been usually based on cultured microorganisms, which may not adequately represent the soil microbial community in either nutrient-poor or in altered carbon and nutrient conditions following fertilisation. We report an alternative approach in which changes in microbial P storage are determined as the residual in a mass balance of extractable P before and after incubation. This approach was applied in three low-fertility sandy soils of southwestern Australia, to determine microbial P immobilisation during 5-day incubations in response to the amendment by 2.323 mg C g⁻¹, 100 μg N g⁻¹ and 20 μg P g⁻¹. The net P immobilisation during the amended incubations determined to be 18.1, 14.1 and 16.3 μg P g⁻¹ in the three soils, accounting for 70.6-90.5% of P added through amendment. Such estimates do not rely on fumigation and K_p values, but for comparison with the FE method we estimated ‘nominal’ K_p values to be 0.20-0.31 for the soils under the amended conditions. Our results showed that microbial P immobilisation was a dominant process regulating P concentration in soil water following the CNP amendment. The mass-balance approach provides information not only about changes in the microbial P compartment, but also about other major P-pools and their fluxes in regulating soil-water P concentrations under substrate- and nutrient-amended conditions.     

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.     

Soil organic matter dynamics and land use change at a grassland/forest ecotone

In the grassland/forest ecotone of North America, many areas are experiencing afforestation and subsequent shifts in ecosystem carbon (C) stocks. Ecosystem scientists commonly employ a suite of techniques to examine how such land use changes can impact soil organic matter (SOM) forms and dynamics. This study employs four such techniques to compare SOM in grassland (Bromus inermis) and recently forested (∼35 year, Ulmus spp. and Quercus spp.) sites with similar soil types and long-term histories in Kansas, USA. The work examines C and nitrogen (N) parameters in labile and recalcitrant SOM fractions isolated via size and density fractionation, acid hydrolysis, and long-term incubations. Size fractionation highlighted differences between grassland and forested areas. N concentration of forested soils’ 63-212 μm fraction was higher than corresponding grassland soils’ values (3.0±0.3 vs. 2.3±0.3 mg g_fraction⁻¹, P<0.05), and N concentration of grassland soils’ 212-2000 μm fraction was higher than forested soils (3.0±0.4 vs. 2.3±0.2 mg g_fraction⁻¹, P<0.05). Similar trends were observed for these same fractions for C concentration; forested soils exhibited 1.3 times the C concentration in the 63-212 μm fraction compared to this fraction in grassland soils. Fractions separated via density separation and acid hydrolysis exhibited no differences in [C], [N], δ¹⁵N, or δ¹³C when compared across land use types. Plant litterfall from forested sites possessed significantly greater N concentrations than that from grassland sites (12.41±0.10 vs. 11.62±0.19 mg g_litter⁻¹). Long-term incubations revealed no differences in C or N dynamics between grassland and forested soils. δ¹³C and δ¹⁵N values of the smallest size and the heavier density fractions, likely representing older and more recalcitrant SOM, were enriched compared to younger and more labile SOM fractions; δ¹⁵N of forested soils’ 212-2000 μm fraction were higher than corresponding grassland soils (1.7±0.3‰ vs. 0.5±0.4‰). δ¹³C values of acid hydrolysis fractions likely reflect preferential losses of ¹³C-depleted compounds during hydrolysis. Though C and N data from size fractions were most effective at exhibiting differences between grassland and forested soils, no technique conclusively indicates consistent changes in SOM dynamics with forest growth on these soils. The study also highlights some of the challenges associated with describing SOM parameters, particularly δ¹³C, in SOM fractions isolated by acid hydrolysis.     

Measuring microbial uptake of nitrogen in nutrient-amended sandy soils—A mass-balance based approach

Soil microbial biomass N is commonly determined through fumigation-extraction (FE), and a conversion factor (K_EN) is necessary to convert extractable N to actual soil biomass N. Estimation of K_EN has been constrained by various uncertainties including potential microbial immobilisation. We developed a mass-balance approach to quantify changes in microbial N storage during nutrient-amended incubation, in which microbial uptake is determined as the residual in a ‘mass-balance’ based on soil-water N before and after amended incubation. The approach was applied to three sandy soils of southwestern Australia, to determine microbial N immobilisation during 5-day incubation in response to supply of 2.323 mg C g⁻¹, 100 μg N g⁻¹ and 20 μg P g⁻¹. The net N immobilisation was estimated to be 95-114 μg N g⁻¹ in the three soils, equivalent to 82.7-85.1% of soil-water N following the amendment. Such estimation for microbial uptake does not depend on fumigation and K_EN conversion, but for comparison purposes we estimated ‘nominal’ K_EN values (0.11-0.14) for the three soils, which were comparable to previously reported K_EN from soils receiving C and N amendment. The accuracy of our approach depends on the mass-balance equation and the integrated measurement errors of the multiple N pools, and was assessed practically through recoveries of added-N when microbial uptake can be minimised. Near-satisfactory recoveries were achieved under such conditions. Our mass-balance approach provides information not only about changes in the microbial biomass nitrogen storage, but also major N-pools and their fluxes in regulating soil N concentrations under substrate and nutrient amended conditions.     

Contribution of primary organic matter to the fatty acid pool in agricultural soils

Fatty acids as major compounds of soil lipids may affect many soil properties, but the input and turnover rates in soil are largely unknown. The objective of this study was to identify and quantify fatty acids in soils as a result of input from primary sources such as plant residues, farmyard manure and soil organisms, and to evaluate the corresponding turnover- and stabilization processes. The concentrations of n-C_10:0 to n-C_34:0 fatty acids were determined in the Ap horizon of a Phaeozem with long-term cropping of rye and maize and the treatments ‘Unfertilized’ (‘U’) and fertilized with ‘Farmyard manure’ (‘FYM’). The most important primary sources of fatty acids such as rye and maize stubble and roots, soil micro- and mesofauna, and the applied FYM were also investigated. The quantification of fatty acids by gas chromatography/mass spectrometry (GC/MS) showed that long-term FYM application led to larger concentrations of n-alkyl fatty acids in the plots grown with rye (‘U’: 48.1 μg g⁻¹, ‘FYM’: 57.7 μg g⁻¹, **P≤0.01, n=3) and maize (‘U’: 17.0 μg g⁻¹, ‘FYM’: 23.4 μg g⁻¹, ***P≤0.001, n=3). The observed bimodal fatty acid distribution in soils from n-C_10:0 to n-C_21:0 and from n-C_21:0 to n-C_34:0 with a predominance at n-C_16:0 and at n-C_28:0 was apparently due to input from crop residues, soil organisms and FYM. The short-chain lengths may have originated from the investigated primary sources. The major contributors to the long-chain lengths, with a maximum at n-C_28:0, were rye stubble and FYM. A change in mono-culture from rye to maize, 38 years prior to sampling, led to a decrease in fatty acid concentrations by factors of about 2.8 (‘U’) and 2.5 (‘FYM’). Therefore, rye-derived fatty acids and soil tillage had a larger impact on fatty acid pools than the input of primary organic matter. The changes in fatty acid distributions and pools under the consideration of the quantified input of primary organic matter led to the conclusion that the short-chained fatty acids were more rapidly decomposed than the long-chains.     

Carbon decomposition in broiler litter-amended soils

Organic carbon (OC) is generally low in Alabama (U.S.A.) soils and varies considerably with cropping systems. Information on decomposition rates of the added C is a prerequisite to designing strategies that improve C sequestration in farming systems. Different models including exponential models have been used to describe OC mineralization in soils as well as to describe its potential as CO₂ to be released into the environment. We investigated the decomposition of broiler litter added to ten non-calcareous soils (Appling, Troup, Cecil, Decatur, Sucarnoochee, Linker, Hartsells, Dothan, Maytag, and Colbert soils). A non-linear regression approach for N mineralization was used to estimate the potentially mineralizable OC pools (C_o) and the first-order rate constant (k) in the soil samples. Results showed that the non-amended soils have distinct differences in their ability to release their native OC as CO₂ and can be divided into four groups depending on their potentially mineralizable C (C_o) and their ability to protect stable organic matter. Sucarnoochee soil represents the first group and contains a moderate amount of OC (11.4 g C kg⁻¹) but had the highest C_o (7.30 g C kg⁻¹ soil). The second distinct group of soils has C_o varying between 5.50 and 5.00 g C kg⁻¹ soil (Decatur, Hartsells, Dothan, and Maytag). The third group has C_o between 5.00 and 4.00 (Appling, Cecil, and Linker). The fourth group has C_o less than 4.00 g C kg⁻¹ soil (Troup and Colbert). Half-life of C remaining in non-amended soils varied from 26 days in Maytag soil to 139 days in Cecil soil. The OC in these non-amended soils represents a very stable form of organic C and thus, not easily decomposed by soil microorganisms. In the broiler litter-amended soils, the C_o varied from 3.82 g C kg⁻¹ in Appling soil amended with broiler litter 1-7.04 g C kg⁻¹ soil in Maytag amended with broiler litter 2. Decomposition of the added OC proceeded in two phases with less than 31% decomposed in 43 days. Potentially mineralizable organic C (C_o) was related to soil organic C (r = 0.661**) and soil C/N ratio (r = 0.819*).     

Soil organic C stabilization and thresholds in C saturation

When building soil organic matter (SOM) contents in agricultural production systems, stabilization of both pre-existing as well as added C is important. A laboratory mineralization experiment was conducted over 374 days to evaluate the effect of pre-existing SOM on soil C mineralization after addition of organic matter (OM) using sugar cane. The SOM gradient used here stretched from 21 to 106 g C kg⁻¹ soil and was a result of different periods of continuous cultivation of 5, 20, 35 and 105 years in comparison to a forest soil. The rate of organic C mineralization was found to be dependent on the status of pre-existing soil organic C (SOC). Highly degraded soil which had been under continuous cultivation for 35 years and more showed the highest rate of C mineralization per unit SOC (117.9 mg C g⁻¹ C) while forest soil had the lowest amount of C mineralized per unit SOC (73.5 mg C g⁻¹ C). Forest soil had the highest amount of increased C mineralization as a result of organic matter (OM) additions (8.0 mg C g⁻¹ soil) followed by the highly degraded soil that had been under cultivation for 105 years (5.5 mg C g⁻¹ soil). Additional mineralized C as a function of time after forest conversion declined progressively within the first 20 years of continuous soil use. Soil which had been under continuous cultivation for 20 years had the lowest amount of additional mineralized C (4.0 mg C g⁻¹ soil). SOM stabilization efficiency in the studied soils appears to be highest with intermediate cultivation history of about 20 years. These soils that have been recently converted to cultivation also appear to have a greater ability to stabilize added OM than the most degraded soils investigated in this study. It is thus advisable to provide intervention strategies to reverse SOM decline for farming communities at an intermediate stage before the soils are highly depleted of SOC.     

Changes in soil organic carbon dynamics in an Eastern Chinese coastal wetland following invasion by a C4 plant Spartina alterniflora

The exotic C₄ grass Spartina alterniflora was intentionally introduced to tidal coastal wetlands in Jiangsu province of China in 1982. Since then it has rapidly replaced the native C₃ plant Suaeda salsa, becoming one of the dominant vegetation types in the coastal wetlands of China. Although plant invasion can change soil organic carbon (SOC) storage, little is known about how plant invasion influences C storage within soil fractions. We investigated how S. alterniflora invasion across an 8, 12 and 14-year chronosequence affected SOC and soil nitrogen (N), using soil fractionation and stable δ¹³C isotope analyses. SOC and N concentrations at 0-10 cm depth in S. alterniflora soil increased during the S. alterniflora invasion chronosequence, ranging from 3.67 to 4.90 g C kg⁻¹ soil, and from 0.307 to 0.391 g N kg⁻¹ soil. These were significantly higher than the values in the Suaeda salsa community, by 27.0-69.6% for SOC, and 21.8-55.2% for total N. The S. alterniflora-derived SOC varied from 0.40 to 0.92 g C kg⁻¹ according to mixing calculations, assuming the two possible SOC sources of S. alterniflora and S. salsa, and accounted for 10.8-18.7% of total SOC in the colonized soils. The estimated accumulative rate of SOC from C₄ (S. alterniflora) was 64.1 C kg⁻¹ soil year⁻¹ and from C₃ sources was 78.1 mg C kg⁻¹. The concentration of S. alterniflora-derived SOC significantly decreased from coarse fraction to fine fraction, and linearly increased as the period of S. alterniflora invasion increased. The highest accumulative rate of SOC from a C₄ source occurred in macroaggregates, while the highest rate from C₃ was in microaggregates. The storage of SOC derived from S. alterniflora in the macroaggregates was 0.27-0.44 g C kg⁻¹ soil, accounting for 43.1-49.1% of the total C₄derived SOC in the soil. Our results suggest that S. alterniflora invasion in coastal wetlands could facilitate SOC storage, because of the high potential for accumulation of the C which has been newly derived from S. alterniflora litter and roots.     

Long-term nitrogen additions increased surface soil carbon concentration in a forest plantation despite elevated decomposition

Forests cover one-third of the Earth’s land surface and account for 30-40% of soil carbon (C). Despite numerous studies, questions still remain about the factors controlling forest soil C turnover. Present understanding of global C cycle is limited by considerable uncertainty over the potential response of soil C dynamics to rapid nitrogen (N) enrichment of ecosystems, mainly from fuel combustion and fertilizer application. Here, we present a 15-year-long field study and show an average increase of 14.6% in soil C concentration in the 0-5 cm mineral soil layer in N fertilized (defined as N+ hereafter) sub-plots of a second-rotation Pinus radiata plantation in New Zealand compared to control sub-plots. The results of ¹⁴C and lignin analyses of soil C indicate that N additions significantly accelerate decomposition of labile and recalcitrant soil C. Using an annual-time step model, we estimated the soil C turnover time. In the N+ sub-plots, soil C in the light (a density < 1.70 g cm⁻³) and heavy fractions had the mean residence times of 23 and 67 yr, respectively, which are lower than those in the control sub-plots (36 and 133 yr in the light and heavy fractions, respectively). The commonly used lignin oxidation indices (vanillic acid to vanillin and syringic acid to syringaldehyde ratios) were significantly greater in the N+ sub-plots than in the control sub-plots, suggesting increased lignin decomposition due to fertilization. The estimation of C inputs to forest floor and δ¹³C analysis of soil C fractions indicate that the observed buildup of surface soil C concentrations in the N+ sub-plots can be attributed to increased inputs of C mass from forest debris. We conclude that long-term N additions in productive forests may increase C storage in both living tree biomass and soils despite elevated decomposition of soil organic matter.     

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

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

Organic matter in density fractions of water-stable aggregates in silty soils: Effect of land use

The location of soil organic matter (SOM) within the soil matrix is considered a major factor determining its turnover, but quantitative information about the effects of land cover and land use on the distribution of SOM at the soil aggregate level is rare. We analyzed the effect of land cover/land use (spruce forest, grassland, wheat and maize) on the distribution of free particulate organic matter (POM) with a density <1.6 g cm⁻³ (free POM_<1.6), occluded particulate organic matter with densities <1.6 g cm⁻³ (occluded POM_<1.6) and 1.6-2.0 g cm⁻³ (occluded POM_1.6-2.0) and mineral-associated SOM (>2.0 g cm⁻³) in size classes of slaking-resistant aggregates (53-250, 250-1000, 1000-2000, >2000 μm) and in the sieve fraction <53 μm from silty soils by applying a combined aggregate size and density fractionation procedure. We also determined the turnover time of soil organic carbon (SOC) fractions at the aggregate level in the soil of the maize site using the ¹³C/¹²C isotope ratio. SOM contents were higher in the grassland soil aggregates than in those of the arable soils mainly because of greater contents of mineral-associated SOM. The contribution of occluded POM to total SOC in the A horizon aggregates was greater in the spruce soil (23-44%) than in the grassland (11%) and arable soils (19%). The mass and carbon content of both the free and occluded POM fractions were greater in the forest soil than in the grassland and arable soils. In all soils, the C/N ratios of soil fractions within each aggregate size class decreased in the following order: free POM_<1.6>occluded POM_<1.6-2.0>mineral-associated SOM. The mean age of SOC associated with the <53 μm mineral fraction of water-stable aggregates in the Ap horizon of the maize site varied between 63 and 69 yr in aggregates >250 μm, 76 yr in the 53-250 μm aggregate class, and 102 yr in the sieve fraction <53 μm. The mean age of SOC in the occluded POM increased with decreasing aggregate size from 20 to 30 yr in aggregates >1000 μm to 66 yr in aggregates <53 μm. Free POM had the most rapid rates of C-turnover, with residence times ranging from 10 yr in the fraction >2000 μm to 42 yr in the fraction 53-250 μm. Results indicated that SOM in slaking-resistant aggregates was not a homogeneous pool, but consisted of size/density fractions exhibiting different composition and stability. The properties of these fractions were influenced by the aggregate size. Land cover/land use were important factors controlling the amount and composition of SOM fractions at the aggregate level.     

High specific activity in low microbial biomass soils across a no-till evapotranspiration gradient in Colorado

The need to identify microbial community parameters that predict microbial activity is becoming more urgent, due to the desire to manage microbial communities for ecosystem services as well as the desire to incorporate microbial community parameters within ecosystem models. In dryland agroecosystems, microbial biomass C (MBC) can be increased by adopting alternative management strategies that increase crop residue retention, nutrient reserves, improve soil structure and result in greater water retention. Changes in MBC could subsequently affect microbial activities related to decomposition, C stabilization and sequestration. We hypothesized that MBC and potential microbial activities that broadly relate to decomposition (basal and substrate-induced respiration, N mineralization, and β-glucosidase and arylsulfatase enzyme activities) would be similarly affected by no-till, dryland winter wheat rotations distributed along a potential evapotranspiration (PET) gradient in eastern Colorado. Microbial biomass was smaller in March 2004 than in November 2003 (417 vs. 231 μg g⁻¹ soil), and consistently smaller in soils from the high PET soil (191 μg g⁻¹) than in the medium and low PET soils (379 and 398 μg g⁻¹, respectively). Among treatments, MBC was largest under perennial grass (398 μg g⁻¹). Potential microbial activities did not consistently follow the same trends as MBC, and the only activities significantly correlated with MBC were β-glucosidase (r = 0.61) and substrate-induced respiration (r = 0.27). In contrast to MBC, specific microbial activities (expressed on a per MBC basis) were greatest in the high PET soils. Specific but not total activities were correlated with microbial community structure, which was determined in a previous study. High specific activity in low biomass, high PET soils may be due to higher microbial maintenance requirements, as well as to the unique microbial community structure (lower bacterial-to-fungal fatty acid ratio and lower 17:0 cy-to-16:1ω7c stress ratio) associated with these soils. In conclusion, microbial biomass should not be utilized as the sole predictor of microbial activity when comparing soils with different community structures and levels of physiological stress, due to the influence of these factors on specific activity.     

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.     

Assaying the effects of chemical ameliorants with earthworms and plants exposed to a heavily polluted metalliferous soil

The study examined the effects of chemical ameliorant additions (1% montmorillonite, 1% hydroxylapatite, or 1% ferrous oxide) on the availability of cadmium (Cd), copper (Cu), lead (Pb) and zinc (Zn) to the earthworm, Lumbricus rubellus, exposed for 4 weeks to a circumneutral heavily polluted soil (Cd = 220 μg g^–1; Cu = 35 μg g^–1; Pb = 6070 μg g^–1; Zn = 124500 μg g^–1) in 1:0–1:3 dilutions with a clean soil, under laboratory conditions. Soil type (i.e. the dilution series) had a strong influence on the 1 M ammonium acetate extractable metal fractions in soil and on worm-tissue concentrations of Cd, Pb and Zn. Soil treatments (i.e. amelioration) significantly reduced only the soil Zn extractable fraction; Zn concentrations in worms tended to be lower in amended soils. A second experiment, involving curly cress (Lepidium sativum), grown either directly in the serial soil dilutions with 5% ameliorant additions or in the water-extractable fractions of the soils, indicated that root growth is a more sensitive endpoint of metal availability than chlorophyll assays. It was concluded that: (i) chemical immobilization of metals is probably most effective in soils with low to moderate degrees of metal pollution; (ii) an integrated suite of bioassays incorporating different, ecologically relevant, taxa is to be recommended for monitoring metal bioavailabilities and biological effects.     

Filtration increases the correlation between water extractable organic carbon and soil microbial activity

A study was carried out in order to establish the relationship between the water extractable organic carbon (WEOC) content of soils and soil microbial activity, and to determine how variations in the extraction procedure might influence the quantity of WEOC recovered. Concentrations of WEOC were determined in soils taken from 12 different sites in the south east of Scotland, using a procedure in which samples were shaken with distilled water, centrifuged at 5000g and then filtered through 0.45 μm Millipore filters. Filtration resulted in between 30 and 400 μg C g⁻¹ being extracted using this procedure and the concentration of WEOC in the resultant extracts correlated with soil microbial production of CO₂ and dehydrogenase activity (P<0.001). Without filtration, although more WEOC was extracted (between 31 and 716 μg C g⁻¹), there was no significant correlation with biological activity. There was also no correlation between WEOC and nitrous oxide release during the incubations. Centrifugation at 20,000g for at least 10 min prior to filtration was required to remove particulate organic materials. Storage of samples at 4 °C or for up to 1 week or freezing for up to 3 months was not found to have a large influence on the concentration of WEOC in extracts, although amounts increased with soil:extractant ratio and increasing extraction time (from 15 to 60 min).     

Rapid quantification of Bacillus subtilis antibiotics in the rhizosphere

Biological control agents like Bacillus subtilis offer an alternative and supplement to synthetic pesticides. Antibiotic production by biocontrol strains of B. subtilis can play a major role in plant disease suppression. Our current understanding of B. subtilis antibiosis comes from culture media measurements of antibiotic production and in vitro suppression of pathogens. Quantifying the antibiotic metabolite chemistry of B. subtilis biofilms growing on root surfaces provides a more accurate understanding of in vivo antibiotic production. An analytical method based on solid-phase extraction (SPE) with high-performance liquid chromatography (HPLC) and mass spectroscopy (MS) has been developed to quantify antibiotics produced by B. subtilis growing on plant roots. Cucumber (Cucumis sativus) was grown in composted soil and potting media inoculated with B. subtilis strain QST 713 (AgraQuest, USA). Two important B. subtilis antibiotics, surfactin and iturin A, were extracted from root and rhizosphere soil using acidified organic solvents followed by cleaning and concentration using SPE. HPLC and HPLC-MS were used to measure surfactin and iturin A. Rhizosphere concentrations of both antibiotics increased with plant age. For plants grown in peat-based potting media, surfactin concentrations increased from 9 μg g⁻¹ root fresh weight (RFW) at 15 d to 30 μg g⁻¹ RFW at 43 d. Iturin concentrations were 7 μg g⁻¹ RFW at 15 d and 180 μg g⁻¹ RFW at 43 d. In an initial field trial in a composted fine sandy loam, we demonstrated rhizosphere production of surfactin and iturin under competition and predation by the myriad macro- and microfauna existing in a fertile high-organic soil, with mature B. subtilis-inoculated cucumber roots yielding 33 μg g⁻¹ RFW surfactin and 630 μg g⁻¹ RFW iturin at 78 d.     

Three-source-partitioning of microbial biomass and of CO2 efflux from soil to evaluate mechanisms of priming effects

We propose and successfully applied a new approach for 3-source-partitioning based on a combination of ¹⁴C labeling with ¹³C natural abundance. By adding ¹⁴C-labeled glucose to soil after C₃ - C₄ vegetation change, we partitioned three C sources in three compartments, namely CO₂, microbial biomass and dissolved organic C (DOC). This enabled us to estimate mechanisms and sources of priming effects (PE).Glucose application at low and high rate (GL: 100 and GH: 1000 μg C g⁻¹, respectively) caused positive PE both short-term (during 1-3 days) and long-term (3-55 days). Despite a 10-fold difference in the amount of substrate added, the PE observed was larger by a factor of only 1.6 at the high versus low rate of glucose. The real and apparent priming effects were distinguished by partitioning of microbial C for glucose-C and SOM-derived C. As the amount of primed CO₂ respired during short-term PE was 40% lower than microbial C, and the contribution of soil C in microbial biomass did not increase, we concluded that such short-term PE was apparent and was mainly caused by accelerated microbial turnover (at GL) and by pool substitution (at GH). Both the amount of primed CO₂-C, which was 1.3-2.1 times larger than microbial C, and the increased contribution of soil C in microbial biomass allowed us to consider the long-term PE as being real. The sole source of real PE (GL treatment) was the “recent” soil organic matter, which is younger than 12-year-old C. The real PE-induced by a glucose amount exceeding microbial biomass (GH) was due to the almost equal contribution of ‘recent’ (<12 years) and ‘old’ (>12 years) C. Thus, the decomposition of old recalcitrant SOM was induced only by an amount of primer exceeding microbial C. We conclude that combining ¹⁴C labeling with ¹³C natural abundance helped disentangle three C sources in CO₂, microbial biomass and DOC and evaluate mechanisms and sources of PE.     

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.     

Heterotrophic and autotrophic nitrification in two acid pasture soils

Laboratory incubation experiments, using ¹⁵N-labeling techniques and simple analytical models, were conducted to measure heterotrophic and autotrophic nitrification rates in two acid soils (pH 4.8-5.3; 1/5 in H₂O) with high organic carbon contents (6.2-6.8% in top 5 cm soil). The soils were from pastures located near Maindample and Ruffy in the Northeast Victoria, Australia. Gross rates of N mineralization, nitrification and immobilization were measured. The gross rates of autotrophic nitrification were 0.157 and 0.119 μg N g⁻¹ h⁻¹ and heterotrophic nitrification rates were 0.036 and 0.009 μg N g⁻¹ h⁻¹ for the Maindample and Ruffy soils, respectively. Heterotrophic nitrification accounted for 19% and 7% of the total nitrification in the Maindample and Ruffy soils, respectively. The heterotrophic nitrifiers used organic N compounds and no as the substrate for nitrification.