Influences of organic fertilization and solarization in a greenhouse on particle-size fractions of a Mediterranean sandy soil

 The effects of a composted organic amendment and solarization on the organic matter (OM) of a sandy soil were determined by means of particle-size fractionation and analysis of carbon and nitrogen contents. After 2 years, total soil carbon increased under organic fertilization but did not significantly change with solarization. As a consequence of the climatic conditions in the greenhouse, the carbon concentrations (g kg^–1 fraction) of the particle-size fractions were lower than those found for temperate soils and closer to those for tropical soils. The carbon amounts (g kg^–1 soil) and carbon:nitrogen ratios, which were highest in fractions >200 μm, reflected the short-term influence of the industrially processed organic amendment, rich in composted coarse plant debris. In contrast, the characteristics of the OM associated with each fraction were not significantly affected by solarization. In comparison with other coarse-textured temperate or tropical soils, carbon concentrations in fine silt (2–20 μm) and clay (0–2 μm) fractions were very low. This suggests a "greenhouse effect", together with a high rate of carbon mineralization affecting fine silt and clay fractions. Received: 19 November 1999     

Studies of nitrogen immobilization and mineralization in calcareous soils—I: Distribution of immobilized nitrogen amongst soil fractions of different particle size and density

¹⁵NO₃^? was immobilized in a calcareous clay and a calcareous sandy soil during incubation of each soil with glucose and wheat straw. Changes in the distribution of immobilized ¹⁵N amongst soil extracts and soil fractions of different particle size and density were determined during periods of net N immobilization.The nature of the organic-C amendment, but not soil type, significantly influenced both the distribution of the immobilized ¹⁵N and the pattern of changes of the organic-¹⁵N of soil fractions with time. In straw-amended soils, approx. 20% of the organic-¹⁵N became associated with a light fraction, sp. gr. < 1.59, the remainder becoming distributed mainly amongst the silt and clay fractions. In glucoseamended soils, very little (< 1.2%) of the ¹⁵N was immobilized in the light fraction, sp. gr. < 1.59, most being rapidly distributed amongst the silt and clay fractions. During a period of complete immobilization, organic-¹⁵N was transferred from the fine clay to the silt and coarse clay fractions.Silt, coarse clay and fine clay components from glucose-amended soils sampled at the end of the net immobilization phase were further fractionated densimetrically into light (sp. gr. < 2.06) and heavy (sp. gr. > 2.06) subfractions. The organic-¹⁵N of respective light subfractions accounted for 43–64% of the total organic-¹⁵N of the silt, 1–9% of that of the coarse clay and 19–21% of that of the fine clay fractions.     

Effect of climate, soil type and earthworm activity on nitrogen transfer from a nitrogen-15-labelled decomposing material under field conditions

 N transfer from ¹⁵N-labelled decomposing material into the microbial biomass and inorganic N forms was studied for more than 2 years at three experimental sites differing in climatic conditions and earthworm abundance. The ¹⁵N-labelled decomposing material was mixed with low-elevation soil (LES), mid-elevation soil (MES) and high-elevation soil (HES). The amended soils were put into two kinds of plastic cylinders closed on both sides with nets preventing (0.1 cm mesh) and allowing (0.5 cm mesh) access by earthworms, and were buried in soil (20 cm depth) to monitor the transfer of N from the ¹⁵N-labelled decomposing material. Climate and soil type play an important role in the release of N from decomposing material. LES transplanted to more humid sites (mid- and high-elevation sites) showed an increase in most of its biological activities (N atom % excess, and microbial biomass C and N). Furthermore, LES was a sandy soil in which the ¹⁵N-labelled decomposing material was less bound than in MES and HES, which contained more silt and clay. This resulted in faster organic matter turnover when climatic conditions were favourable. The presence of earthworms greatly increased the quantity of inorganic N (mainly NH₄ ⁺) in the soils and enhanced the release of N from the ¹⁵N-labelled decomposing material and the native organic matter, compared to soil without earthworms. Received: 21 January 1999     

Quantification of nitrogen excretion rates for three lumbricid earthworms using 15N

 Nitrogen excretion rates of ¹⁵N-labeled earthworms and contributions of ¹⁵N excretion products to organic (dissolved organic N) and inorganic (NH₄-N, NO₃-N) soil N pools were determined at 10  °C and 18  °C under laboratory conditions. Juvenile and adult Lumbricus terrestris L., pre-clitellate and adult Aporrectodea tuberculata (Eisen), and adult Lumbricus rubellus (Hoffmeister) were labeled with ¹⁵N by providing earthworms with ¹⁵N-labeled organic substrates for 5–6 weeks. The quantity of ¹⁵N excreted in unlabeled soil was measured after 48 h, and daily N excretion rates were calculated. N excretion rates ranged from 274.4 to 744 μg N g^–1 earthworm fresh weight day^–1, with a daily turnover of 0.3–0.9% of earthworm tissue N. The N excretion rates of juvenile L. terrestris were significantly lower than adult L. terrestris, and there was no difference in the N excretion rates of pre-clitellate and adult A. tuberculata. Extractable N pools, particularly NH₄-N, were greater in soils incubated with earthworms for 48 h than soils incubated without earthworms. Between 13 and 40% of excreted ¹⁵N was found in the ¹⁵N-mineral N (NH₄-N+NO₃-N) pool, and 13–23% was in the ¹⁵N-DON pool. Other fates of excreted ¹⁵N may have been incorporation in microbial biomass, chemical or physical protection in non-extractable N forms, or gaseous N losses. Earthworm excretion rates were combined with earthworm biomass measurements to estimate N flux from earthworm populations through excretion. Annual earthworm excretion was estimated at 41.5 kg N ha^–1 in an inorganically-fertilized corn agroecosystem, and was equivalent to 22% of crop N uptake. Our results suggest that the earthworms could contribute significantly to N cycling in corn agroecosystems through excretion processes. Received: 12 April 1999     

Nitrogen distribution and 15N natural abundances in particle size fractions of a long‐term agricultural field experiment

The present study combined a physical fractionation procedure with the determination of the natural abundance of ¹⁵N to investigate the impact of organic manure and mineral fertilizer application, and fallow on changes of N associated with different soil particle size fractions. The long‐term field experiment was conducted since 1956 in Ultuna, Sweden, on an Eutric Cambisol. Nitrogen in bulk soil and in particle size fractions changed significantly since 1956. The N_t concentrations in bulk soil decreased in all treatments not receiving organic materials. Comparing the N contribution of particle‐size fractions to the total N amount revealed the following ranking: silt > clay > fine clay > fine sand > coarse sand. The relative contribution of N in silt sized particles significantly increased from low to high bulk soil N contents, whereas N in clay and fine clay fractions decreased. The C : N ratios of particle size fractions differed considerably more between treatments than C : N ratios in bulk soils. Generally, the C : N ratios decreased from coarse to fine fractions emphasizing the tendency of smaller fractions being more significant as N sink than as C_org sink. ¹⁵N abundances varied more between particle size fractions of single treatments than between bulk soil from differently treated plots. Within treatments we observed differences of up to 7.1 ‰ between particle size fractions. In most cases δ ¹⁵N values increased with decreasing particle sizes. This pattern on average was similar to changes in δ ¹³ C. Our results suggest that silt sized particles acted as medium‐term sink of introduced N and that ¹⁵N abundances in particle size fractions sensitively reflect changes in N status in response to soil management.     

Effect of drying and rewetting on mineralization and distribution of bacterial constituents in soil fractions

Summary Mineralization of ¹⁴C- and ¹⁵N-labelled whole bacteria, cytoplasm, and cell walls and their distribution in different soil fractions were studied during 211 days of incubation including two drying and rewetting cycles. With any of these three soil amendments, almost 60% of C derived from cellular constituents was released as CO₂, 15% was incorporated into the living microbial biomass and 25% was distributed into protected microbial metabolites or recalcitrant microbial products. The distribution of C and N derived from the amendments in the different soil fractions showed that constituents adsorbed on fine clay (<0.2 m were more rapidly decomposed than those adsorbed on silt (50-2 ) and coarse clay (2–0.2 ), indicating a faster organic matter turnover in fine clay than in silt and coarse clay. Although alternate soil drying and rewetting cycles did not significantly affect the mineralization of bacterial constituents, the cycles did have an important effect on the size and specific activities of newly formed microbial biomass. This suggests the presence of an active and a dormant fraction of soil biomass.     

Influence of earthworm activity on aggregate-associated carbon and nitrogen dynamics differs with agroecosystem management

Earthworms are known to be important regulators of soil structure and soil organic matter (SOM) dynamics, however, quantifying their influence on carbon (C) and nitrogen (N) stabilization in agroecosystems remains a pertinent task. We manipulated population densities of the earthworm Aporrectodea rosea in three maize-tomato cropping systems [conventional (i.e., mineral fertilizer), organic (i.e., composted manure and legume cover crop), and an intermediate low-input system (i.e., alternating years of legume cover crop and mineral fertilizer)] to examine their influence on C and N incorporation into soil aggregates. Two treatments, no-earthworm versus the addition of five A. rosea adults, were established in paired microcosms using electro-shocking. A ¹³C and ¹⁵N labeled cover crop was incorporated into the soil of the organic and low-input systems, while ¹⁵N mineral fertilizer was applied in the conventional system. Soil samples were collected during the growing season and wet-sieved to obtain three aggregate size classes: macroaggregates (>250 μm), microaggregates (53-250 μm) and silt and clay fraction (<53 μm). Macroaggregates were further separated into coarse particulate organic matter (cPOM), microaggregates and the silt and clay fraction. Total C, ¹³C, total N and ¹⁵N were measured for all fractions and the bulk soil. Significant earthworm influences were restricted to the low-input and conventional systems on the final sampling date. In the low-input system, earthworms increased the incorporation of new C into microaggregates within macroaggregates by 35% (2.8 g m⁻² increase; P=0.03), compared to the no-earthworm treatment. Within this same cropping system, earthworms increased new N in the cPOM and the silt and clay fractions within macroaggregates, by 49% (0.21 g m⁻²; P<0.01) and 38% (0.19 g m⁻²; P=0.02), respectively. In the conventional system, earthworms appeared to decrease the incorporation of new N into free microaggregates and macroaggregates by 49% (1.38 g m⁻²; P=0.04) and 41% (0.51 g m⁻²; P=0.057), respectively. These results indicate that earthworms can play an important role in C and N dynamics and that agroecosystem management greatly influences the magnitude and direction of their effect.     

Mineral-fixed ammonium in clay- and silt-size fractions of soils incubated with 15N-ammonium sulphate for five years

Summary Four soils with 6, 12, 23, and 47% of clay were incubated for 5 years with ¹⁵N-labeled (NH_{4 2}SO₄ and hemicellulose. The incubations took place at 20°C and 55% water-holding capacity. Samples of whole soils, and clay- (<2 m) and silt-(2–20 m) size fractions (isolated by ultrasonic dispersion and gravity sedimentation) were analysed for labeled and native mineral-fixed ammonium. Mineral-fixed ammonium in non-incubated soil samples accounted for 3.4%–8.3% of the total N and showed a close positive correlation with the soil clay content (r ² = 0.997). After 5 years of incubation, the content of mineral-fixed ammonium in the clay fraction was 255–430 g N g^–1, corresponding to 71%–82% of the mineral-fixed ammonium in whole soils. Values for silt were 72–166 g N g^–1 (14%–33% of whole soil content). In the soils with 6% and 12% clay, less than 1 % of the labeled clay N was present as mineral-fixed ammonium. In the soil with 23% clay, 3% of the labeled N in the clay was mineral-fixed ammonium. Labeled mineral-fixed ammonium was not detected in the silt fractions. For whole soils, and clay and silt fractions, the proportion of native N present as mineral-fixed ammonium varied between 3% and 6%. In contrast, the proportion of labeled N found as mineral-fixed ammonium in the soil with 4701o clay was 23%, 38% and 31% for clay, silt, and whole-soil samples, respectively. Corresponding values for native mineral-fixed ammonium were 12%, 16%, and 10%. Consequently, studies based on soil particle-size fractions and addressing the N turnover in clay-rich soils should consider the pool of mineral-fixed ammonium, especially when comparing results from different size fractions with those from fractions isolated from soils of a widely different textural composition.     

Transformations and recovery of residue and fertilizer nitrogen-15 in a sandy Lixisol of West Africa

 The fate of ¹⁵N-labeled plant residues from different cover-cropping systems and labeled inorganic N fertilizer in the organic, soil mineral, microbial biomass and soil organic matter (SOM) particle-size fractions was investigated in a sandy Lixisol. Plant residues were from mucuna (legume), lablab (legume), imperata (grass), maize (cereal) and mixtures of mucuna or lablab with imperata or maize, applied as a surface mulch. Inorganic N fertilizer was applied as ¹⁵N-(NH₄)₂SO₄ at two rates (21 and 42 mg N kg^–1 soil). Total N release from mucuna or lablab residues was 2–3 times higher than from the other residues, whereas imperata immobilized N throughout the study period. In contrast, ¹⁵N was mineralized from all the plant residues irrespective of the mineralization–immobilization pattern observed for total N. After 168 days, 69% of soil mineral N in mucuna- or lablab-mulched soils was derived from the added residues, representing 4–8% of residue N, whereas 9–30% of inorganic N was derived from imperata, maize and the mixed residues. At the end of the study, 4–19% of microbial biomass N was derived from the added residue/fertilizer-N, accounting for 1–3% of added residue-N. Averaged across treatments, particulate SOM fractions accounted for less than 1% of the total soil by weight but contained 20% of total soil C and 8% of soil N. Soils amended with mucuna or lablab incorporated more N in the 250–2000 μm SOM pool, whereas soil amended with imperata or the mixed residues incorporated similar proportions of labeled N in the 250–2000 μm and 53–250 μm fractions. In contrast, in soils receiving the maize or inorganic fertilizer-N treatments, higher proportions of labeled N were incorporated into the 53–250 μm than the 250–2000 μm fractions. The relationship between these differences in residue/fertilizer-N partitioning into different SOM particle-size fractions and soil productivity is discussed. Received: 12 March 1999     

THE DISTRIBUTION OF LABELLED SUGARS IN SOIL PARTICLE SIZE FRACTIONS AS A MEANS OF DISTINGUISHING PLANT AND MICROBIAL CARBOHYDRATE RESIDUES

Soils of the Countesswells and Insch series incubated with ¹⁴C labelled glucose or plant materials have been separated into clay (< 2 μm), silt, (2–20 μm), fine sand (20–250 μm) and coarse sand (>250μm) fractions and the distribution of individual labelled and unlabelled sugars was determined in each fraction. Both soils contained about 10–15 per cent clay, 18–23 per cent silt and about 60 per cent fine and coarse sand. For all soil samples the concentrations of sugars were usually greatest in the clay, slightly less in the silt, with values in the sand fractions being five or ten times lower, except when fresh plant material was present. In ¹⁴C glucose amended Insch soil, 55 per cent of the radioactivity in sugars (predominantly hexoses) occurred in the clay, 36 per cent in the silt, 3 per cent in the fine sand and 6 per cent in the coarse sand after 28 days incubation. For the Countesswells soil the values were 55, 42, 2 and 1 per cent respectively. In ¹⁴C ryegrass amended soil before incubation. 77 per cent of the radioactivity in sugars (predominantly glucose, arabinose and xylose) was in the coarse sand. After one year's incubation this had fallen to 59 per cent. In soil amended with ¹⁴C cereal rye straw the distribution of radioactivity in sugars after four years incubation was: clay, 21 per cent; silt, 43 per cent; fine sand, 21 per cent; coarse sand, 4 per cent. These distributions were compared with that of the naturally occurring sugars: clay, 31–42 per cent; silt, 40–43 per cent; fine sand, 3–11 per cent; coarse sand, 12–20 per cent.     

Studies of nitrogen immobilization and mineralization in calcareous soils—II: Mineralization of immobilized nitrogen from soil fractions of different particle size and density

¹⁵NO^?₃ was immobilized in a calcareous sandy soil and a calcareous clay soil each incubated with glucose and wheat straw. Net mineralization of organic-¹⁵N was more rapid in the sandy soil, irrespective of C amendment, and in soils amended with glucose. Intermittent drying and wetting of soils during incubation stimulated mineralization of ¹⁵N-labelled and native soil organic-N in all treatments. The availability (percentage mineralization) of recently-immobilized ¹⁵N consistently exceeded that of the native soil N. Ratios of the availability of labelled and unlabelled N were similar in the sandy and clay soils but varied according to C amendment, drying and wetting cycle and incubation period.Changes in the distribution of immobilized N amongst soil extracts and soil fractions of different particle size and density were determined during periods of net N mineralization. In straw-amended soils, the organic-¹⁵N of a light fraction, sp.gr. < 1.59, decomposed relatively rapidly during the late mineralization period. Decreases of organic ¹⁵N of the fine clay fraction were also recorded. In glucose-amended soils, net N mineralization was accompanied by significant decreases in the concentrations of organic-¹⁵N of the silt and fine clay fractions.Drying and rewetting of soils hastened or magnified changes occurring in the organic-¹⁵N of soil fractions, but qualitatively, the pattern of change was similar to that observed with soils incubated under uniformly-moist conditions.The percentage distribution of labelled and unlabelled N suggested that in the long term, the silt fraction will accumulate an increasing proportion of the more stable nitrogenous residues.     

Soil organic matter fractionation as a tool for predicting nitrogen mineralization in silty arable soils

After decades of searching for a practical method to estimate the N mineralization capacity of soil, there is still no consistent methodology. Indeed it is important to have practical methods to estimate soil nitrogen release for plant uptake and that should be appropriate, less time consuming, and cost effective for farmers. We fractionated soil organic matter (SOM) to assess different fractions of SOM as predictors for net N mineralization measured from repacked (disturbed) and intact (undisturbed) soil cores in 14 weeks of laboratory incubations. A soil set consisting of surface soil from 18 cereal and root‐cropped arable fields was physically fractionated into coarse and fine free particulate OM (coarse fPOM and fine fPOM), intra‐microaggregate particulate OM (iPOM) and silt and clay sized OM. The silt and clay sized OM was further chemically fractionated by oxidation with 6% NaOCl to isolate an oxidation‐resistant OM fraction, followed by extraction of mineral bound OM with 10% HF (HF‐res OM). Stepwise multiple linear regression yielded a significant relationship between the annual N mineralization (kg N/ha) from undisturbed soil and coarse fPOM N (kg N/ha), silt and clay N (kg N/ha) and its C:N ratio (R² = 0.80; P < 0.01). The relative annual N mineralization (% of soil N) from disturbed soils was related to coarse fPOM N, HF‐res OC (% of soil organic carbon) and its C:N ratio (R² = 0.83; P < 0.01). Physical fractions of SOM were thus found to be the most useful predictors for estimating the annual N mineralization rate of undisturbed soils. However, the bioavailability of physical fractions was changed due to the disturbance of soil. For disturbed soils, a presumed stable chemical SOM fraction was found to be a relevant predictor indicating that this fraction still contains bio‐available N. The latter prompted a revision in our reasoning behind selective oxidation and extraction as tools for characterizing soil organic N quality with respect to N availability. Nonetheless, the present study also underscores the potential of a combined physical and chemical fractionation procedure for isolating and quantifying N fractions which preferentially contribute to bulk soil N mineralization. The N content or C:N ratio of such fractions may be used to predict N mineralization in arable soils.     

Effects of long-term compost and fertilizer application on stability of aggregate-associated organic carbon in an intensively cultivated sandy loam soil

The study examined the influence of compost and mineral fertilizer application on the content and stability of soil organic carbon (SOC). Soil samples collected from a long-term field experiment were separated into macroaggregate, microaggregate, and silt + clay fractions by wet-sieving. The experiment involved seven treatments: compost, half-compost N plus half-fertilizer N, fertilizer NPK, fertilizer NP, fertilizer NK, fertilizer PK, and control. The 18-year application of compost increased SOC by 70.7–121.7%, and mineral fertilizer increased by 5.4–25.5%, with no significant difference between control soil and initial soil. The C mineralization rate (rate per unit dry mass) in microaggregates was 1.52–2.87 mg C kg⁻¹ day⁻¹, significantly lower than in macroaggregate and silt + clay fractions (P < 0.05). Specific C mineralization rate (rate per unit SOC) in silt + clay fraction amounted to 0.48–0.87 mg C g⁻¹ SOC day⁻¹ and was higher than in macroaggregates and microaggregates. Our data indicate that SOC in microaggregates is more stable than in macroaggregate and silt + clay fractions. Compost and mineral fertilizer application increased C mineralization rate in all aggregates compared with control. However, compost application significantly decreased specific C mineralization rate in microaggregate and silt + clay fractions by 2.6–28.2% and 21.9–25.0%, respectively (P < 0.05). By contrast, fertilizer NPK application did not affect specific C mineralization rate in microaggregates but significantly increased that in silt + clay fractions. Carbon sequestration in compost-amended soil was therefore due to improving SOC stability in microaggregate and silt + clay fractions. In contrast, fertilizer NPK application enhanced SOC with low stability in macroaggregate and silt + clay fractions.     

Soil colloids-bound plasmid DNA: Effect on transformation of E. coli and resistance to DNase I degradation

The adsorption and binding of plasmid p34S DNA on four different colloidal fractions from a Brown soil and clay minerals in the presence of various Ca²⁺ concentrations, the ability of bound DNA to transform competent cells of CaCl₂-treated Escherichia coli, and the resistance of bound DNA to degradation by DNase I were studied. DNA adsorption on soil colloids and clay minerals was promoted in the presence of Ca²⁺. Kaolinite exhibited the highest adsorption affinity for DNA among the examined soil colloids and clay minerals. In comparison with organo-mineral complexes (organic clays) and fine clays (<0.2 μm), DNA was tightly adsorbed by H₂O₂-treated clays (inorganic clays) and coarse clays (0.2-2 μm). The transformation efficiency of bound DNA increased with increasing concentrations of Ca²⁺ at which soil colloid or clay mineral-DNA complexes were formed. DNA bound by kaolinite showed the lowest transformation efficiency, and especially no transformants were observed with kaolinite-DNA complex prepared at 5-100 mM Ca²⁺. Compared to organic clays and fine clays, DNA bound on inorganic clays and coarse clays showed a lower capacity to transform E. coli at different Ca²⁺ concentrations. The presence of soil colloids and minerals provided protection to DNA against degradation by DNase I. Montmorillonite, organic clays and fine clays showed stronger protective effects for DNA than inorganic clays and coarse clays. The protection mechanisms as well as the differences in transforming efficiency of plasmid DNA molecules bound on various soil colloidal particles are discussed. The information obtained in this study is of fundamental significance for the understanding of the horizontal dissemination of recombinant DNA and the fate of extracellular DNA in soil environments.     

Structure and function of the soil microbial community in microhabitats of a heavy metal polluted soil

 Particle-size fractionation of a heavy metal polluted soil was performed to study the influence of environmental pollution on microbial community structure, microbial biomass, microbial residues and enzyme activities in microhabitats of a Calcaric Phaeocem. In 1987, the soil was experimentally contaminated with four heavy metal loads: (1) uncontaminated controls; (2) light (300 ppm Zn, 100 ppm Cu, 50 ppm Ni, 50 ppm V and 3 ppm Cd); (3) medium; and (4) heavy pollution (two- and threefold the light load, respectively). After 10 years of exposure, the highest concentrations of microbial ninhydrin-reactive nitrogen were found in the clay (2–0.1 μm) and silt fractions (63–2 μm), and the lowest were found in the coarse sand fraction (2,000–250 μm). The phospholipid fatty acid analyses (PLFA) and denaturing gradient gel electrophoresis (DGGE) separation of 16S rRNA gene fragments revealed that the microbial biomass within the clay fraction was predominantly due to soil bacteria. In contrast, a high percentage of fungal-derived PLFA 18 : 2ω6 was found in the coarse sand fraction. Bacterial residues such as muramic acid accumulated in the finer fractions in relation to fungal residues. The fractions also differed with respect to substrate utilization: Urease was located mainly in the <2 μm fraction, alkaline phosphatase and arylsulfatase in the 2–63 μm fraction, and xylanase activity was equally distributed in all fractions. Heavy metal pollution significantly decreased the concentration of ninhydrin-reactive nitrogen of soil microorganisms in the silt and clay fraction and thus in the bulk soil. Soil enzyme activity was reduced significantly in all fractions subjected to heavy metal pollution in the order arylsulfatase >phosphatase >urease >xylanase. Heavy metal pollution did not markedly change the similarity pattern of the DGGE profiles and amino sugar concentrations. Therefore, microbial biomass and enzyme activities seem to be more sensitive than 16S rRNA gene fragments and microbial amino-sugar-N to heavy metal treatment. Received: 21 January 2000     

Nitrogen dynamics in different types of pasture in the Austrian Alps

 Soil N dynamics were compared in Alpine pastures on two mountains. N-pool sizes and N fluxes were measured relative to N losses via leaching and denitrification in summer. On each mountain, four types of pasture were studied: (1) forest pastures, (2) recently developed pastures formed by forest clearance ("new pastures"), (3) older established pastures, and (4) pastures planted with clover. At both study sites (Scheuchegg and Teufelstein) we obtained similar results. Compared with forest pasture soils, open pasture soils were found to have greater microbial biomass and faster mineralisation potentials, but net field mineralisation rates were slower. In the forest pastures, highest N losses via denitrification were found. Higher potential leaching of NO₃ ^–, estimated by accumulation of NO₃ ^– on ion-exchange resins, in the forest pasture soils suggests lower N uptake by microbes and herbaceous plants compared with open pastures. N₂O-production rates of the forest pasture soils at the Scheuchegg site (11.54 μg N₂O-N m^–2 h^–1) were of similar magnitude to those reported for spruce forests without pastures, but at Teufelstein (53.75 μg N₂O-N m^–2 h^–1) they were higher. However, if forest pastures are not overgrazed, no elevated N loss through N₂O production and leaching of NO₃ ^– is expected. Denitrification rates in the open pastures (0.83–7.50 μg N₂O-N m^–2 h^–1) were low compared with reports on lowland pastures. In soils of the new pastures, rates of microbial N processes were similar to those in the established pastures, indicating a high capacity of soils to restore their internal N cycle after forest clearance. Received: 19 August 1999     

Post-fire mineral N allocation and stabilisation in soil particle size fractions in Mediterranean grassland and shrubland

In the Mediterranean region, long-term post-fire soil N dynamics may be relevant in the stabilisation of soil organic matter and N cycling in the plant-soil system. Post-fire recycling of N may be retained by the retention of N in physically and/or chemically protected fractions of soil organic matter. We studied the allocation of post-fire ¹⁵N-tracer among different soil organic matter fractions (coarse sand, fine sand, coarse silt and fine silt + clay) and ¹⁵N-tracer dynamics for 12 years after prescribed fires in three different Mediterranean plant communities (grassland, mixed shrub-grassland and shrubland). We selected 6 plots for each community and we set experimental fires. Directly after the fires, we applied ¹⁵NH₄⁺-N and we monitored the fate of ¹⁵N-tracer over a period of 12 years. For this purpose, we carried out a physical size fractionation and we analysed the biochemical recalcitrance of N and ¹⁵N by acid hydrolysis in the size fractions obtained. In both burned and unburned plots the finest soil particles (<20 μm) accounted for most soil N. Fire promoted N increases in the medium size fractions while the N pool in the finest and coarsest fractions did not change after the fires. Interestingly, ¹⁵N-tracer was quickly incorporated into fine fractions from which, in the case of plant communities free of legumes, it was remobilised in the following years. Fire did not promote changes in recalcitrant N, but shrubland showed marked decreases in N recalcitrance 6 years after the fires. Despite the fact that the primary effects on soil fractions were detected just after the fires, these persisted for 12 years post-fire. Newly incorporated ¹⁵N-tracer was less recalcitrant than total N and, surprisingly, fine fractions had very low recalcitrant ¹⁵N values, similar to the coarse fractions. Apparently, the N transformations in the finest fraction (<20 μm) were mainly regulated by the quality of the ¹⁵N compounds retained in the fraction.     

Characterization of organic matter in topsoils under rain forest and pasture in the eastern Brazilian Amazon basin

 In topsoils under forest and 7-, 12- and 17-year-old pastures, organic matter was characterized by analysing C and N distribution in particle-size fractions, the C decomposition rates of soil and particle-size fractions and by employing density-fractionation of macro-organic matter (>150 μm). The C and N associated with clay fractions increased with increasing age of pasture. The weight (%) of macro-organic matter and its heavy fractions (>1.37 g cm^–3) also increased with increasing age of pasture. However, in a long-term incubation (100 days), these changes seemed to involve an increase in the C decomposition rate in the topsoil of the oldest pasture. Using the C decomposition rates of particle-size fractions, it appeared that silt and clay contributed differently to C decomposition in the whole soil. C associated with silt contributed to the C decomposition rate during the first 40 days of incubation, while C associated with clay contributed to C decomposition in the long-term incubation (after 40 days), especially when the clay fraction appeared to reach saturation point with respect to its ability to bind organic compounds and thus protect the soil from C loss. Received: 13 March 1998     

Effects of animal manure and mineral fertilizer on the total carbon and nitrogen contents of soil size fractions

Summary Soil was sampled in autumn 1984 in the 132 field (sandy loam soil) of the Askov long-term experiments (started in 1894) and fractionated according to particle size using ultrasonic dispersion and sedimentation in water. The unmanured plot and plots given equivalent amounts of N (1923–1984 annual average, 121 kg N/ha) in either animal manure or mineral fertilizer were sampled to a depth of 15 cm, fractionated and analysed for C and N. Mineral fertilizer and animal manure increased the C and N content of whole soil, clay (<2 m) and silt (2–20 m) size fractions relative to unmanured samples, while the C content of the sand size fractions (fine sand 1, 20–63 m; fine sand 2, 63–200 m; coarse sand, 200–2000 m) was less affected. Clay contained 58% and 65°70 of the soil C and N, respectively. Corresponding values for silt were 30% and 26%, while sand accounted for 10% of the soil C. Fertilization did not influence this distribution pattern. The C : N ratio of the silt organic matter (14.3) was higher and that of clay (10.6) lower than whole-soil C:N ratios (12.0). Fertilization did not influence clay and silt C : N ratios. Animal manure caused similar relative increases in the organic matter content of clay and silt size fractions (36%). In contrast, mineral fertilizer only increased the organic matter content of silt by 21% and that of clay by 14%.     

Phospholipid fatty acid profiles and xylanase activity in particle size fractions of forest soil and casts of Lumbricus terrestris L. (Oligochaeta,Lumbricidae)

Earthworms are key agents in organic matter decomposition, as they remove surface plant litter material and mix it with mineral soil. Plant litter material is comminuted in the gizzard of anecic earthworms and this is enhanced if sand particles are available. We hypothesize that this comminution of soil and litter will result in changes in the distribution of soil organic matter and soil microorganisms in the different particle-size fractions. We investigated soil organic matter content, xylanase- and microbial activity and community structure in bulk soil and particle size fractions of Lumbricus terrestris L. casts and in soil with and without the addition of beech litter.Earthworm gut passage did not affect the particle-size distribution but the content of soil organic matter was decreased in the fine sand fraction in treatments without litter (−6.80%) and increased in treatments with litter (+33.23%). The soil organic matter content of the clay fraction tended to be higher in earthworm casts. Xylanase activity was at a maximum in the fine sand fraction, lower in the coarse sand fraction and at a similar minimum in the silt- and clay-sized fraction. In the coarse sand fraction of the cast and litter treatments xylanase activity was increased by 39.1% and 124.8%, respectively. In the silt-sized fraction of casts the addition of litter increased xylanase activity (+58.6%) whereas, in casts without litter it was decreased (−36.25%). In the particle size fractions of casts, the content of bacterial PLFAs was decreased in the fine sand fraction and tended to be decreased in the clay fraction compared to the respective fractions in soil. In the silt fraction the fungal-to-bacterial PLFA ratio was higher in casts than in soil.We conclude that earthworms stabilize soil organic matter in cast aggregates predominantly by increasing the soil organic matter content in the clay fraction where it becomes protected against microbial attack. Organic matter in the coarse and fine sand fractions is decomposed primarily by fungi; xylanase is very active in these sand fractions and incorporation of litter into these fractions by the earthworms increased fungal biomass. Comminution of litter during passage through the earthworm gut increased the biomass and activity of fungi also in the silt fraction. The use of PLFA profiles in combination with other quantitative microbial methods improves the understanding of stabilizing and mobilizing processes in earthworm casts.