Gross N transformation rates and net N mineralisation rates related to the C and N contents of soil organic matter fractions in grassland soils of different age

We investigated the relationship between soil organic matter (SOM) content and N dynamics in three grassland soils (0-10 and 10-20 cm depth) of different age (6, 14 and 50 y-old) with sandy loam textures. To study the distribution of the total C and N content the SOM was fractionated into light, intermediate and heavy density fractions of particulate macro-organic matter (150-2000 μm) and the 50-150 μm and <50 μm size fractions. The potential gross N transformation rates (mineralisation, nitrification, NH₄⁺ and NO₃⁻ immobilization) were determined by means of short-term, fully mirrored ¹⁵N isotope dilution experiments (7-d incubations). The long-term potential net N mineralisation and gross N immobilization rates were measured in 70-d incubations. The total C and N contents mainly tended to increase in the 0-10 cm layer with increasing age of the grassland soils. Significant differences in total SOM storage were detected for the long-term (50 y-old) conversion from arable land to permanent grassland. The largest relative increase in C and N contents had occurred in the heavy density fraction of the macro-organic matter, followed by the 50-150 and <50 μm fractions. Our results suggest that the heavy density fraction of the macro-organic matter could serve as a good indicator of early SOM accumulation, induced by converting arable land to permanent grassland. Gross N mineralisation, nitrification, and (long-term) gross N immobilization rates tended to increase with increasing age of the grasslands, and showed strong, positive correlations with the total C and N contents. The calculated gross N mineralisation rates (7-d incubations) and net N mineralisation rates (70-d incubations) corresponded with a gross N mineralisation of 643, 982 and 1876 kg N ha⁻¹ y⁻¹, and a net N mineralisation of 195, 208 and 274 kg N ha⁻¹ y⁻¹ in the upper 20 cm of the 6, 14 and 50 y-old grassland soils, respectively. Linear regression analysis showed that 93% of the variability of the gross N mineralisation rates could be explained by variation in the total N contents, whereas total N contents together with the C-to-N ratios of the <50 μm fraction explained 84% of the variability of the net N mineralisation rates. The relationship between long-term net N mineralisation rates and gross N mineralisation rates could be fitted by means of a logarithmic equation (net m=0.24Ln(gross m)+0.23, R²=0.69, P<0.05), which reflects that the ratio of gross N immobilization-to-gross N mineralisation tended to increase with increasing SOM contents. Microbial demand for N tended to increase with increasing SOM content in the grassland soils, indicating that potential N retention in soils through microbial N immobilization tends to be limited by C availability.     

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.     

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.     

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.     

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

Microbial biomass N dynamics were studied under field and laboratory conditions in soils of high yield (HY) and low yield (LY) areas in an agricultural field. The objective of the study was to determine the size and activity of soil microbial biomass in the soils of the different yield areas and to compare these data obtained under field and laboratory conditions. Soils were amended with ¹⁵N labelled mustard (Sinapis alba) residues (both experiments) and labelled nitrate (laboratory only) at 30 μg N g⁻¹ dry soil. Soil microbial biomass (SMB) N, mineral N (N_min) and total N content was monitored both in the field and in the laboratory. N₂O efflux was additionally measured in laboratory treatments. Isotope ratios were determined for SMB in both experiments, for all other parameters only in the laboratory treatments. In the laboratory less amounts of added substrate N were immobilised by the SMB in HY soils compared to LY soils, whereas in the field immobilisation of added N by SMB was higher in HY soils initially and slightly lower after 40 days of incubation. Calculated turnover times in the laboratory nitrate, laboratory mustard and field mustard amendments were 0.18, 0.27 and 0.74 years (HY) and 0.22, 0.61 and 1.01 years (LY), respectively. The turnover times of added substrate N always showed the trend to be faster in HY soils compared to LY soils. A faster turnover of nutrients in the HY soils may involve a better nutrient supply of the plants, which coincides with the higher agricultural yield observed in these areas.     

Shifts in amino sugar and ergosterol contents after addition of sucrose and cellulose to soil

An incubation experiment with organic soil amendments was carried out with the aim to determine whether formation and use of microbial tissue (biomass and residues) could be monitored by measuring glucosamine and muramic acid. Living fungal tissue was additionally determined by the cell-membrane component ergosterol. The organic amendments were fibrous maize cellulose and sugarcane sucrose adjusted to the same C/N ratio of 15. In a subsequent step, spherical cellulose was added without N to determine whether the microbial residues formed initially were preferentially decomposed. In the non-amended control treatment, ergosterol remained constant at 0.44 μg g⁻¹ soil throughout the 67-day incubation. It increased to a highest value of 1.9 μg g⁻¹ soil at day 5 in the sucrose treatment and to 5.0 μg g⁻¹ soil at day 33 in the fibrous cellulose treatment. Then, the ergosterol content declined again. The addition of spherical cellulose had no further significant effects on the ergosterol content in these two treatments. The non-amended control treatment contained 48 μg muramic acid and 650 μg glucosamine g⁻¹ soil at day 5. During incubation, these contents decreased by 17% and 19%, respectively. A 33% increase in muramic acid and an 8% increase in glucosamine were observed after adding sucrose. Consequently, the ratio of fungal C to bacterial C based on bacterial muramic acid and fungal glucosamine was lowered in comparison with the other two treatments. No effect on the two amino sugars was observed after adding cellulose initially or subsequently during the second incubation period. This indicates that the differences in quality between sucrose and cellulose had a strong impact on the formation of microbial residues. However, the amino sugars did not indicate a preferential decomposition of microbial residues as N sources.     

The contribution of soil organic matter fractions to carbon and nitrogen mineralization and microbial community size and structure

The aims of this study were to: (i) assess the impact of hay and fertilizer application on organic matter (OM) fractions (dissolved organic matter (DOM), light fraction organic matter (LFOM, <1.0 g cm⁻³), heavy fraction OM (HFOM, <1.7 g cm⁻³)), carbon (C) and nitrogen (N) cycling processes and microbial community size and structure, and (ii) quantify the role of OM fractions to C and N cycling. Soil was collected in 2001 from a field experiment to which grass hay (1996) and/or fertilizer (1995 and 1999) had previously been applied. DOM-C (P<0.05) and DOM-N (P=0.07) were significantly higher in control and fertilized soil than hay and hay+fertilized soil. LFOM and HFOM C and N contents and C/N ratios were significantly (P<0.05) higher in hay+fertilized and hay amended soil than in control and fertilized soil. Potentially mineralizable-N (PMN), microbial biomass-C (MB-C), microbial biomass-N (MB-N) and microbial respiration (CO₂) were not affected by fertilizer and/or hay application. Gross N mineralization (Gross Min) and gross nitrification (Gross Nit) rates were significantly (P<0.05) higher in fertilized, hay, hay+fertilized soil than control soil. However, there was no significant difference between treatments in gross N immobilization rates. Results reported here highlight the importance of a labile fraction of the DOM pool to N and C cycling as its removal significantly (P<0.05) reduced PMN, MB-N, Gross Min and Gross Nit compared with whole soil in most or all treatments. In soil where DOM+LFOM were removed PMN was significantly (P<0.05) lower, but MB-C, Gross Min and Gross Nit was significantly (P<0.05) higher than in DOM removed soil. This suggests that LFOM plays an important role as a sink for mineral-N. Total soil phospholipid fatty acid (PLFA) concentration was significantly (P<0.05) higher in hay amended than control, fertilized and hay+fertilized soil. Principal components analysis was able to clearly discriminate between control, fertilized, hay+fertilized and hay amended soil. Soil amended with hay or fertilizer had a microbial community structure which differed from that of the control or hay+fertilized soils. Redundancy analysis with Monte Carlo permutation tests revealed that PLFA profiles were strongly correlated to differences in Gross Min, Gross Nit, MB-N, MB-C, MB-C/N ratio, total soil C and total soil C/N ratio. The results of this research suggest that changes in microbial structure are related to aspects of soil C and N pools and cycling.     

Initial recovery of soil carbon and nitrogen pools and dynamics following disturbance in jack pine forests: A comparison of wildfire and clearcut harvesting

Forests naturally maintained by stand-replacing wildfires are often managed with clearcut harvesting, yet we know little about how replacing wildfire with clearcutting affects soil processes and properties. We compared the initial recovery of carbon (C) and nitrogen (N) pools and dynamics following disturbance in jack pine (Pinus banksiana) stands in northern Lower Michigan, USA, by sampling soils (Oa+A horizons) from three “treatments”: 3-6-year-old harvest-regenerated stands, 3-6-year-old wildfire-regenerated stands and 40-55-year-old intact, mature stands (n=4 stands per treatment). We measured total C and N; microbial biomass and potentially mineralizable C and N; net nitrification; and gross rates of N mineralization and nitrification. Burned stands exhibited reduced soil N but not C, whereas clearcut and mature stands had similar quantities of soil organic matter. Both disturbance types reduced microbial biomass C compared to mature stands; however, microbial biomass N was reduced in burned stands but not in clearcut stands. The experimental C and N mineralization values were fit to a first-order rate equation to estimate potentially mineralizable pool size (C₀ and N₀) and rate parameters. Values for C₀ in burned and clearcut stands were approximately half that of the mature treatment, with no difference between disturbance types. In contrast, N₀ was lowest in the wildfire stands (170.2 μg N g⁻¹), intermediate in the clearcuts (215.4 μg N g⁻¹) and highest in the mature stands (244.6 μg N g⁻¹). The most pronounced difference between disturbance types was for net nitrification. These data were fit to a sigmoidal growth equation to estimate potential NO₃⁻ accumulation (Nit_max) and kinetic parameters. Values of Nit_max in clearcut soils exceeded that of wildfire and mature soils (149.2 vs. 83.5 vs. 96.5 μg NO₃⁻-N g⁻¹, respectively). Moreover, the clearcut treatment exhibited no lag period for net NO₃⁻ production, whereas the burned and mature treatments exhibited an approximate 8-week lag period before producing appreciable quantities of NO₃⁻. There were no differences between disturbances in gross rates of mineralization or nitrification; rather, lower NO₃⁻ immobilization rates in the clearcut soils, 0.20 μg NO₃⁻ g⁻¹ d⁻¹ compared to 0.65 in the burned soils, explained the difference in net nitrification. Because the mobility of NO₃⁻ and NH₄⁺ differs markedly in soil, our results suggest that differences in nitrification between wildfire and clearcutting could have important consequences for plant nutrition and leaching losses following disturbance.     

Priming effect of biuret addition on native soil N mineralisation under laboratory conditions

This study was carried out to quantify the priming effect of biuret on native soil nitrogen (N) mineralisation during a 112-day incubation. Addition of biuret (100 mg ¹⁵N-labelled biuret kg⁻¹ soil) increased the turnover rate constant of soil organic matter and had a positive priming effect on native soil N mineralisation in two soils. The additional mineralisation was 0.65% of the total soil N (equivalent to 47.1 kg N ha⁻¹) in a sandy loam soil and 0.62% of the soil N (equivalent to 46.5 kg N ha⁻¹) in a silt loam soil.     

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.     

Gross fluxes of nitrogen in grassland soil exposed to elevated atmospheric pCO2 for seven years

Plant response to increasing atmospheric CO₂ partial pressure (pCO₂) depends on several factors, one of which is mineral nitrogen availability facilitated by the mineralisation of organic N. Gross rates of N mineralisation were examined in grassland soils exposed to ambient (36 Pa) and elevated (60 Pa) atmospheric pCO₂ for 7 years in the Swiss Free Air Carbon dioxide Enrichment experiment. It was hypothesized that increased below-ground translocation of photoassimilates at elevated pCO₂ would lead to an increase in immobilisation of N due to an excess supply of energy to the roots and rhizosphere. Intact soil cores were sampled from Lolium perenne and Trifolium repens swards in May and September, 2000. The rates of gross N mineralisation (m) and NH₄⁺ consumption (c) were determined using ¹⁵N isotopic dilution during a 51-h period of incubation. The rates of N immobilisation were estimated either as the difference between m and the net N mineralisation rate or as the amount of ¹⁵N released from the microbial biomass after chloroform fumigation. Soil samples from both swards showed that the rates of gross N mineralisation and NH₄⁺ consumption did not change significantly under elevated pCO₂. The lack of a significant effect of elevated pCO₂ on organic N turnover was consistent with the similar size of the microbial biomass and similar immobilisation of applied ¹⁵N in the microbial N pool under ambient and elevated pCO₂. Rates of m and c, and microbial ¹⁵N did not differ significantly between the two sward types although a weak (p<0.1) pCO₂ by sward interaction occurred. A significantly larger amount of NO₃⁻ was recovered at the end of the incubation in soil taken from T. repens swards compared to that from L. perenne swards. Eleven percent of the added ¹⁵N were recovered in the roots in the cores sampled under L. perenne, while only 5% were recovered in roots of T. repens. These results demonstrate that roots remained a considerable sink despite the shoots being cut at ground level prior to incubation and suggest that the calculation of N immobilisation from gross and net rates of mineralisation in soils with a high root biomass does not reflect the actual immobilisation of N in the microbial biomass. The results of this study did not support the initial hypothesis and indicate that below-ground turnover of N, as well as N availability, measured in short-term experiments are not strongly affected by long-term exposure to elevated pCO₂. It is suggested that differences in plant N demand, rather than major changes in soil N mineralisation/immobilisation, are the long-term driving factors for N dynamics in these grassland systems.     

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.     

Sulphur immobilization and availability in soils assessed using isotope dilution

Increasing recognition of S deficiency in soils has raised the need for understanding processes governing S cycling and availability in soils. However, the quantification of the two main processes of S cycling, i.e. mineralization and immobilization, remains difficult as these processes occur simultaneously. A modified isotope ³⁵SO₄ dilution technique was developed and used to measure the effect of sulphate (SO₄) fertilization on S mineralization and immobilization in planted (pot experiment with ryegrass (Lolium multiflorum L.)) and unplanted soils (incubation). The immobilization and mineralization of S was calculated from the dynamics of stable and labelled S in soil KH₂PO₄ extracts containing an anion exchange membrane that concentrates SO₄ and mainly excludes other S species. The mathematical analysis of the isotope dilution data differs from methods proposed earlier. The radiolabile S in unplanted soil (E value) and in ryegrass (L value) were used as a measure of total available S in soils. Sulphate immobilization rate significantly declined during incubation. Sulphate application reduced gross mineralization but surprisingly reduced SO₄ immobilization. The E value significantly increased during the incubation in all soils as a result of gross mineralization, e.g. from 3.8 mg S kg⁻¹ at day 0 to 11.5 mg S kg⁻¹ at day 43 in the sandy soil with no sulphate addition. A full recovery in the E value of S added in (+S) treatments was achieved. Similarly, radiolabile S in the above-ground ryegrass biomass (L value) increased with S addition, with a full recovery of added S. The E and L values nearly fit a 1:1 line suggesting identical S dynamics in a planted and unplanted soil. The method proposed has operational advantages compared to methods used earlier.     

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.     

Soil biochemical activity and phosphorus transformations and losses from acidified forest soils

Three Bohemian Forest catchments, Plešné, ?erné and ?ertovo, were studied. These catchments have similar climatic conditions, relief and vegetation, but differ in their bedrock composition. The granitic bedrock in the Plešné catchment was more susceptible to phosphorus (P) leaching under acid conditions than was the mica schist bedrock in the other catchments. The goal of this study was to determine if higher P leaching from the Plešné catchment was associated with differences in microbial P transformations and enzymatic P hydrolysis. Phosphorus and nitrogen contents in soil microbial biomass (P_MB, N_MB; chloroform fumigation), C mineralisation rate (C_min; CO₂ production by GC) and phosphatase activity (MUF-phosphate), were measured in three successive years. Phosphatase activity, P_MB, and C_min were used to characterise the enzymatic hydrolysis of organic P, microbial P accumulation, and microbial mineralisation rates of organic compounds, respectively. Soil chemical properties were characterised by C, N and P content, pH, and by oxalate-extractable P, Fe and Al. Spatial variability in N_MB, P_MB, C_min and phosphatase activity within the catchment was higher (coefficient of variation, CV<50%) than their temporal variability (CV<30%). Multivariate analysis revealed a significant soil layer effect but not that of catchment. When soil layers were evaluated separately, a difference between the Plešné and ?erné or ?ertovo catchments was found in litter and mineral layers, even though the variability within one catchment was high. Within soil profile, phosphatase activity was positively correlated with C_tot, N_MB and C_min (r²=0.89-0.92) being very correlated with P_MB (r²=0.99). Phosphatase activity was higher in the litter (14.0 nmol g⁻¹ h⁻¹) and humus (8.65 nmol g⁻¹ h⁻¹) layers of Plešné than in the same layers of the ?erné (9.65 and 6.40 nmol g⁻¹ h⁻¹) and ?ertovo (12.8 and 6.0 nmol g⁻¹ h⁻¹) soils. Similarly, P_MB in the litter and humus layers of Plešné soil (161 and 93 μg g⁻¹) was higher than P_MB of the same layers of the ?erné (120 and 66 μg g⁻¹) and ?ertovo (148 and 89 μg g⁻¹) soils. High MUFP hydrolysis rate: C_min molar ratio (0.16-1.17 M of P per 1 M of respired C) indicated that potential enzymatic P hydrolysis exceeded estimated microbial P demand (0.034 M of P per 1 M of respired C) in all catchments. The results suggest that higher microbial P transformations and enzymatic P hydrolysis could contribute to enhanced P leaching from the Plešné catchment, which could be enhanced by the lower Fe content in the soil of this catchment as compared to the ?erné and ?ertovo catchments.     

Contribution of nitrification and denitrification to N2O emissions from urine patches

Urine deposition by grazing livestock causes an immediate increase in nitrous oxide (N₂O) emissions, but the responsible mechanisms are not well understood. A nitrogen-15 (¹⁵N) labelling study was conducted in an organic grass-clover sward to examine the initial effect of urine on the rates and N₂O loss ratio of nitrification (i.e. moles of N₂O-N produced per moles of nitrate produced) and denitrification (i.e. moles of N₂O produced per moles of N₂O+N₂ produced). The effect of artificial urine (52.9 g N m⁻²) and ammonium solution (52.9 g N m⁻²) was examined in separate experiments at 45% and 35% water-filled pore space (WFPS), respectively, and in each experiment a water control was included. The N₂O loss derived from nitrification or denitrification was determined in the field immediately after application of ¹⁵N-labelled solutions. During the next 24 h, gross nitrification rates were measured in the field, whereas the denitrification rates were measured in soil cores in the laboratory. Compared with the water control, urine application increased the N₂O emission from 3.9 to 42.3 μg N₂O-N m⁻² h⁻¹, whereas application of ammonium increased the emission from 0.9 to 6.1 μg N₂O-N m⁻² h⁻¹. In the urine-affected soil, nitrification and denitrification contributed equally to the N₂O emission, and the increased N₂O loss resulted from a combination of higher rates and higher N₂O loss ratios of the processes. In the present study, an enhanced nitrification rate seemed to be the most important factor explaining the high initial N₂O emission from urine patches deposited on well-aerated soils.     

Performance of para-nitrophenyl phosphate and 4-methylumbelliferyl phosphate as substrate analogues for phosphomonoesterase in soils with different organic matter content

Phosphomonoesterase (PMEase) activity plays a key role in nutrient cycling and is a potential indicator of soil condition and ecosystem stress. We compared para-nitrophenyl phosphate (pNPP) and 4-methylumbelliferyl phosphate (MUP) as substrate analogues for PMEase in 7 natural ecosystem soils and 8 agricultural top soils with contrasting C contents (8.0-414 g kg⁻¹ C) and pH (3.0-7.5). PMEase activities obtained with pNPP (0.05-5 μmol g⁻¹ h⁻¹) were significantly less than activities obtained with MUP (0.9-13 μmol g⁻¹ h⁻¹), especially in soils with a high organic matter content (>130 g kg⁻¹). Only PMEase activities assayed with MUP correlated significantly with total C and total N (r=0.7, P<0.01 all), and pH (r=−0.71, P<0.01). PMEase activities obtained with the two substrate analogues were correlated when expressed on a C-content basis (r=0.8, P<0.001), but not when expressed on an oven-dry soil weight basis. This indicated that interference by organic matter is related to the quantity rather than to the quality of organic matter. Overall, assaying with MUP was more sensitive compared to assaying with pNPP, particularly in the case of high organic and acid soils.     

Food choice and reproductive success of Folsomia candida feeding on copper-contaminated mycelium of the soil fungus Alternaria alternata

We examined collembolan food preference for fungal mycelium grown on copper-contaminated medium, and the relationship between copper content, food selectivity and collembolan fitness when fed contaminated mycelium.To clarify whether collembolan food selectivity is related to fitness parameters, Folsomia candida were fed mycelium of the dark-pigmented fungus Alternaria alternata grown on medium with different copper concentrations. Copper-contaminated food (fungus grown on 50, 125, 250 and 500 μg Cu g⁻¹ medium, fresh wt.) was offered together with untreated food for 4 weeks. F. candida fed selectively on the provided mycelium and discriminated clearly between mycelium grown on high and low levels of contamination, distinctly preferring fungus grown on medium with a total copper concentration of 50 and 125 μg g⁻¹. In contrast, fungus grown on highly contaminated medium (250 and 500 μg g⁻¹) was avoided. Collembolan food preference generally matched fitness parameters. Reproduction was significantly affected by the total copper concentration of the fungal growth medium. When fed their preferred mycelium, collembolan reproduction was enhanced, whereas a diet of highly contaminated mycelium (250 or 500 μg g⁻¹) resulted in a strong decrease in reproduction. Adult survival was affected only marginally. Even though heavy metal contamination is a potential stress factor for many soil microarthropods, F. candida is able to discriminate between high and low quality food sources, and even benefits from moderately elevated copper concentrations.     

Changes in soil biological and biochemical characteristics in a long-term field trial on a sub-tropical inceptisol

Soil organic carbon (SOC), microbial biomass carbon (MBC), their ratio (MBC/SOC) which is also known as microbial quotient, soil respiration, dehydrogenase and phosphatase activities were evaluated in a long-term (31 years) field experiment involving fertility treatments (manure and inorganic fertilizers) and a maize (Zea mays L.)-wheat (Triticum aestivum L.)-cowpea (Vigna unguiculata L.) rotation at the Indian Agricultural Research Institute near New Delhi, India. Applying farmyard manure (FYM) plus NPK fertilizer significantly increased SOC (4.5-7.5 g kg⁻¹), microbial biomass (124-291 mg kg⁻¹) and microbial quotient from 2.88 to 3.87. Soil respiration, dehydrogenase and phosphatase activities were also increased by FYM applications. The MBC response to FYM+100% NPK compared to 100% NPK (193 vs. 291 mg kg⁻¹) was much greater than that for soil respiration (6.24 vs. 6.93 μl O₂ g⁻¹ h⁻¹) indicating a considerable portion of MBC in FYM plots was inactive. Dehydrogenase activity increased slightly as NPK rates were increased from 50% to 100%, but excessive fertilization (150% NPK) decreased it. Acid phosphatase activity (31.1 vs. 51.8 μg PNP g⁻¹ h⁻¹) was much lower than alkali phosphatase activity (289 vs. 366 μg PNP g⁻¹ h⁻¹) in all treatments. Phosphatase activity was influenced more by season or crop (e.g. tilling wheat residue) than fertilizer treatment, although both MBC and phosphatase activity were increased with optimum or balanced fertilization. SOC, MBC, soil respiration and acid phosphatase activity in control (no NPK, no manure) treatment was lower than uncultivated reference soil, and soil respiration was limiting at N alone or NP alone treatments.