Reproducibility of a soil organic carbon fractionation method to derive RothC carbon pools

Fractionation of soil is undertaken to isolate organic carbon with distinct functional properties, such as stability and turnover times. Soil organic carbon (SOC) fractionation helps us to understand better the response of SOC to changes in land use, management or climate. However, fractionation procedures are often poorly defined and there is little information available on their reproducibility in different laboratories. In a ring trial, we assessed the reproducibility of a SOC fractionation method introduced by Zimmermann et al. (2007). The isolated fractions were linked to the model pool sizes of the Rothamsted carbon model (RothC). We found significant differences between six laboratories for all five defined fractions in three different soils with coefficients of variation ranging from 14 to 138%. During ultrasonic dispersion, the output power (energy per unit time) was identified as an important factor controlling the distribution of SOC within these five fractions, while commonly only the output energy is standardized. The amount of water used to wet‐sieve dispersed soil slurry significantly influenced the amount of extracted dissolved organic carbon (DOC). We therefore suggest using a fixed amount of power for ultrasonic dispersion (20 W) and a minimum amount of water for wet sieving (2000 ml). RothC pool sizes were predicted from the measured fractions and compared with RothC equilibrium pool size distributions. This model initialization using measured SOC fractions, however, led to an over‐estimation of stable RothC SOC pools when compared with pool size distributions derived from RothC equilibrium runs under a bare fallow soil model simulation. To improve the isolation of particulate organic matter from stable mineral‐bound organic matter, we suggest that the density should be increased from 1.8 to 2.0 g cm^?3 in the density fractionation step. We formulated a modified fractionation procedure, which aims specifically to enhance reproducibility across laboratories and to improve the match of the isolated SOC fractions with RothC's SOC pools.     

Separation of light and heavy organic matter fractions in soil — Testing for proper density cut-off and dispersion level

Density fractionation is frequently applied to separate soil organic matter according to the degree and the mode of interaction with minerals. Density fractions are operationally defined by density cut-off and sonication intensity, which determine the nature of the separated material. However, no tests or general agreements exist on the most appropriate density cut-off as well as on method and intensity of dispersion. Numerous variants have been proposed and applied, with results often contrasting each other and being hard to interpret. Here, we aimed at separating two light fractions (free and occluded into aggregates) composed of almost pure organic material, and one heavy fraction comprising the organic–mineral associations. We tested effects of different density cut-offs and sonication intensities, in combination and separately, on fraction yields, as well as on the fractions' organic carbon, total nitrogen and lignin-derived phenols. We tried to find optimum density cut-offs and sonication intensities, providing light fractions with maximum organic material and minimum contamination by mineral material. Under the test conditions, a density of 1.6 g cm^?3 gave best results for all test soils, allowing for separation of maximums amounts of almost pure organic material. The density cut-off at 1.6 g cm^?3 is well in line with previous studies and theoretical considerations, therefore we recommend the use of this density as most suitable for separation of organic debris. Sonication levels for aggregate disruption to achieve complete separation of occluded light organic matter varied amongst soils. The necessary intensity of dispersion relates to the type of soil, depending on the stability of contained aggregates. The application of one single dispersion energy level to different soils may result either in mineral contamination or in incomplete separation of light and heavy fractions as well as in redistribution of organic material amongst fractions. This means there is no single sonication level that can be applied to all soils. Thus, obtaining a meaningful light fraction residing within aggregates (occluded light fraction) requires assessment of the dispersion energy necessary to disrupt the aggregate system of a given soil without dispersion of organic–mineral associations. This can be done in pre-experiments where the soil is fractionated at different sonication levels. The appropriate dispersion is determined by mass yields and OC content of the obtained occluded fractions.     

Redistribution of particulate organic matter during ultrasonic dispersion of highly weathered soils

The use of ultrasonic energy for the dispersion of aggregates in studies of soil organic matter (SOM) fractionation entails a risk of redistribution of particulate organic matter (POM) to smaller particle‐size fractions. As the mechanical strength of straw also decreases with increasing state of decomposition, it can be expected that not all POM will be redistributed to the same extent during such dispersion. Therefore, we studied the redistribution of POM during ultrasonic dispersion and fractionation as a function of (i) dispersion energy applied and (ii) its state of decomposition. Three soils were dispersed at different ultrasonic energies (750, 1500 and 2250 J g^?1 soil) or with sodium carbonate and were fractionated by particle size. Fraction yields were compared with those obtained with a standard particle‐size analysis. Undecomposed or incubated (for 2, 4 or 6 months) ¹³C‐enriched wheat straw was added to the POM fraction (0.25–2 mm) of one of the soils before dispersion and fractionation. Dispersion with sodium carbonate resulted in the weakest dispersion and affected the chemical properties of the fractions obtained through its high pH and the introduction of carbonate. The mildest ultrasonic dispersion treatment (750 J g^?1) did not result in adequate soil dispersion as too much clay was still recovered in the larger fractions. Ultrasonic dispersion at 1500 J g^?1 soil obtained a nearly complete dispersion down to the clay level (0.002 mm), and it did not have a significant effect on the total amount of carbon and nitrogen in the POM fractions. The 2250 J g^?1 treatment was too destructive for the POM fractions since it redistributed up to 31 and 37%, respectively, of the total amount of carbon and nitrogen in these POM fractions to smaller particle‐size fractions. The amount of ¹³C‐enriched wheat straw that was redistributed to smaller particle‐size fractions during ultrasonic dispersion at 1500 J g^?1 increased with increasing incubation time of this straw. Straw particles incubated for 6 months were completely transferred to smaller particle‐size fractions. Therefore, ultrasonic dispersion resulted in fractionation of POM, leaving only the less decomposed particles in this fraction. The amounts of carbon and nitrogen transferred to the silt and clay fractions were, however, negligible compared with the total amounts of carbon and nitrogen in these fractions. It is concluded that ultrasonic dispersion seriously affects the amount and properties of POM fractions. However, it is still considered as an acceptable and appropriate method for the isolation and study of SOM associated with silt and clay fractions.     

Effects of compost on the chemical composition of SOM in density and aggregate fractions from rice–wheat cropping systems as shown by solid‐state 13C‐NMR spectroscopy

The 4‐year application of pig‐manure compost (PMC) to crop fields in Jiangsu significantly increased organic‐C and total N concentrations compared to chemical fertilization and control treatment. To identify the soil processes that led to these changes, ¹³C cross‐polarization magic‐angle spinning nuclear‐magnetic resonance (¹³C CPMAS NMR) and dipolar‐dephasing nuclear‐magnetic‐resonance spectroscopy (DD NMR) were conducted on soil organic matter (SOM) fractions separated by wet‐sieving and density fractionation procedures. This allowed characterization of the SOM quality under three contrasting fertilizer regimes. The results indicate that PMC application can alter the distribution of functional groups and improve alkyl C‐to‐O‐alkyl C ratios compared to chemical‐fertilizer treatment (CF). Alkyl C contents were increased from macroaggregate fractions (> 2 mm) to microaggregate fractions (0.05–0.25 mm) for all treatments, suggesting that recalcitrant material accumulates in the microaggregate fractions. The O‐alkyl C contents were decreased from macroaggregate fractions (> 2 mm) to microaggregate fractions (0.05–0.25 mm) under CF and PMC treatments, while no consistent trend was found for the control (NF) treatment. The alkyl C‐to‐O‐alkyl C ratios in macroaggregates were lower than those in microaggregates, indicating that the degrees of SOM decomposition were lower in macroaggregates compared to microaggregates. In all aggregate‐size classes, the amount of organic matter appeared to depend on the fertilization regime. This study provides useful information regarding the buildup of organic material in soil from long‐term manure‐compost enrichment.     

Effect of ultrasonic power on soil organic carbon fractions

Ultrasonic dispersion is a prevalent tool for soil fractionation. It is widely ignored that variation in ultrasonic power might lead to significantly different dispersion. We evaluated the effect of power variation with constant energy on the fine fraction mass, its organic C content and quality. All parameters increased significantly with power. The term “stable aggregates” as used in fractionation schemes cannot be defined by ultrasonic energy alone but power needs to be standardized, too.     

Density fractionation of organic matter in dolomite‐derived soils

Dolomite (CaMg(CO₃)₂) constitutes half of the global carbonates. Thus, many calcareous soils have been developing rather from dolomitic rocks than from calcite (CaCO₃)‐dominated limestone. We developed a physical fractionation procedure based on three fractionation steps, using sonication with subsequent density fractionation to separate soil organic matter (SOM) from dolomite‐derived soil constituents. The method avoids acidic pretreatment for destruction of carbonates but aims at separating out carbonate minerals according to density. The fractionation was tested on three soils developed on dolostone parent material (alluvial gravel and solid rock), differing in organic‐C (OC) and inorganic‐C (IC) concentrations and degree of carbonate weathering. Soil samples were suspended and centrifuged in Na‐polytungstate (SPT) solutions of increasing density, resulting in five different fractions: two light fractions < 1.8 g cm^–3 (> 20 μm and < 20 μm), rich in OC and free of carbonate, and two organomineral fractions (1.8–2.4 g cm^–3 and 2.4–2.6 g cm^–3), containing 66–145 mg g^–1 and 16–29 mg g^–1 OC. The organomineral fractions consist of residual clay from carbonate weathering such as clay minerals and iron oxides associated with SOM. The fifth fraction (> 2.6 g cm^–3) was dominated by dolomite (85%–95%). The density separation yielded fractions differing in mineral compositions, as well as in SOM, indicated by soil‐type‐specific OC distributions and decreasing OC : N ratios with increasing density of fractions. The presented method is applicable to a wide range of dolomitic and most likely to all other calcareous soils.     

Influence of fertilization and organic amendments on organic‐carbon fractions in Heilu soil on the loess plateau of China

The influence of fertilization on organic‐carbon fractions separated by density and particle size in Heilu soil (Calcic Kastanozems, FAO) was investigated in a 20‐year (1979–1999) long‐term experiment on the Loess Plateau of China. Compared to an unfertilized treatment, N application alone did not increase total organic carbon (TOC) and its fractions of density and particle size. However, the treatment of N + P fertilization significantly increased salty‐solution–soluble organic carbon (SSOC), microbial biomass C (MB‐C), and organic C associated with fine silt. When manure was applied alone and in combination with N and P fertilizer, the light fraction of organic C (LFOC), SSOC, and MB‐C were increased significantly, and the TOC was as high as that of a native Heilu soil. Organic C associated with different particle‐size fractions was also increased significantly, and the allocation of C among the fractions was altered: the proportions of C in sand (>50 μm), coarse‐silt (20–50 μm), and fine‐clay (<0.2 μm) fractions were increased whereas fine‐silt (2–20 μm) and coarse‐clay (0.2–2 μm) fractions were decreased. It is concluded that N fertilizer alone is not capable of restoring organic‐matter content in the Heilu soils of the Loess Plateau and that C‐containing material like manure and straw is necessary to produce significant increase in soil organic carbon in these soils.     

Storage and stability of organic matter and fossil carbon in a Luvisol and Phaeozem with continuous maize cropping: A synthesis

Quantitative information about the amount and stability of organic carbon (OC) in different soil organic‐matter (OM) fractions and in specific organic compounds and compound‐classes is needed to improve our understanding of organic‐matter sequestration in soils. In the present paper, we summarize and integrate results performed on two different arable soils with continuous maize cropping (a) Stagnic Luvisol with maize cropping for 24 y, b) Luvic Phaeozem with maize cropping for 39 y) to identify (1) the storage of OC in different soil organic‐matter fractions, (2) the function of these fractions with respect to soil‐OC stabilization, (3) the importance and partitioning of fossil‐C deposits, and (4) the rates of soil‐OC stabilization as assessed by compound‐specific isotope analyses. The fractionation procedures included particle‐size fractionation, density fractionation, aggregate fractionation, acid hydrolysis, different oxidation procedures, isolation of extractable lipids and phospholipid fatty acids, pyrolysis, and the determination of black C. Stability of OC was determined by ¹³C and ¹⁴C analyses. The main inputs of OC were plant litter (both sites) and deposition of fossil C likely from coal combustion and lignite dust (only Phaeozem).     

Does Particulate Organic Matter Fraction Meet the Criteria for a Model Soil Organic Matter Pool?

There is a well-recognized need for improved fractionation methods to partition soil organic matter into functional pools. Physical separation based on particle size is widely used, yielding particulate organic matter(POM, i.e., free or "uncomplexed" organic matter 50 μm) as the most labile fraction. To evaluate whether POM meets criteria for an ideal model pool, we examined whether it is:1) unique, i.e., found only in the 50 μm fraction and 2) homogeneous, rather than a composite of different subfractions. Following ultrasonic dispersion, sand( 50 μm) along with coarse(20–50 μm) and fine(5–20 μm) silt fractions were isolated from a silt loam soil under long-term pasture at Lincoln, New Zealand. The sand and silt fractions contained 20% and 21% of total soil C, respectively.We adopted a sequential density separation procedure using sodium polytungstate with density increasing step-wise from 1.7 to 2.4 g cm~(-3) to recover organic matter(light fractions) from the sand and silt fractions. Almost all(ca. 90%) the organic matter in the sand fraction and a large proportion(ca. 60%–70%) in the silt fractions was recovered by sequential density separation. The results suggested that POM is a composite of organo-mineral complexes with varying proportions of organic and mineral materials. Part of the organic matter associated with the silt fractions shared features in common with POM. In a laboratory bio-assay, biodegradability of POM varied depending on land use(pasture arable cropping). We concluded that POM is neither homogeneous nor unique.     

Characterization of recently 14C pulse-labelled carbon from roots by fractionation of soil organic matter

The inability of physical and chemical techniques to separate soil organic matter into fractions that have distinct turnover rates has hampered our understanding of carbon (C) and nutrient dynamics in soil. A series of soil organic matter fractionation techniques (chemical and physical) were evaluated for their ability to distinguish a potentially labile C pool, that is ‘recent’ root and root‐derived soil C. ‘Recent’ root and root‐derived C was operationally defined as root and soil C labelled by ¹⁴CO₂ pulse labelling of rye grass–clover pasture growing on undisturbed cores of soil. Most (50–94%) of total soil + root ¹⁴C activity was recovered in roots. Sequential extraction of the soil + roots with resin, 0.1 m NaOH and 1 m NaOH allocated ‘recent’ soil + root ¹⁴C to all fractions including the alkali‐insoluble residual fraction. Approximately 50% was measured in the alkali‐insoluble residue but specific activity was greater in the resin and 1 m NaOH fractions. Hot 0.5 m H₂SO₄ hydrolysed 80% of the ¹⁴C in the alkali‐insoluble residue of soil + roots but this diminished specific activity by recovering much non‐¹⁴C organic matter. Pre‐alkali extraction treatment with 30% H₂O₂ and post‐alkali treatment extractions with hot 1 m HNO₃ removed organic matter with a large ¹⁴C specific activity from the alkali‐insoluble residue. Density separation failed to isolate a significant pool of ‘recent’ root‐derived ¹⁴C. The density separation of ¹⁴C‐labelled roots, and roots remixed with non‐radioactive soil, showed that the adhesion of soil particles to young ¹⁴C‐labelled roots was the likely cause of the greater proportion of ¹⁴C in the heavy fraction. Simple chemical or density fractionations of C appear unsuitable for characterizing ‘recent’ root‐derived C into fractions that can be designated labile C (short turnover time).     

Recovery and dynamics of decomposing plant residue in soil: an evaluation of three fractionation methods

Our goals in this study were to track the incorporation of plant residue into soil organic matter (SOM) and test the effectiveness of different fractionation methods to evaluate this transformation. We incubated soil amended with ¹³C‐labelled barley (Hordeum vulgare L.) residue and used three fractionation methods based on size (> 250, 53–250, 5–53 and < 5 µm) and density (< 1.7 g cm^?3, i.e. light fraction (LF)) and determined its quantity and the rate of C loss or gain or both in these fractions as decomposition progressed. One method was based on size only, another involved density separation followed by size fractionation and a third separated organic matter fractions by size first and then by density. There were significant quantitative differences between the methods for the amount of residue in the fractions, but there was no effect of fractionation method on the rate of change in the residue that comprised the fractions. The density method did not appear to identify all of the most recently added (i.e. least decomposed) residue in the LF or that there was a redistribution of SOM among the fractions. The amount of residue C and the C:N ratio of the residue in the two smallest fractions increased early during the incubation (0–2 months), but subsequently decreased towards the end. The initially small C:N ratio in the clay fraction probably reflects the accumulation of microbial by‐products from the rapid decomposition of water‐soluble compounds. The subsequent increase and decrease in both residue C and C:N ratio reflects the balance of the accumulation of sorbed water‐soluble compounds and dense plant residue fragments and their mineralization over time. We conclude that clay is a sink for residue C (i.e. microbial metabolites) early during decomposition, and that there is a transfer among fractions and mineralization of residue C as decomposition proceeds. These findings indicate that the clay fraction contains a dynamic pool of C that can cycle within short time‐scales.     

Content and composition of free and occluded particulate organic matter in a differently textured arable Cambisol as revealed by solid‐state 13C NMR spectroscopy

The composition of functional light soil organic matter pools of arable Cambisols with a gradient in clay content was investigated. Soil texture differences originate from increasing loess admixture to the parent material (coarse‐grained tertiary sediments). Using density fractionation in combination with ultrasonic dispersion, two types of particulate organic matter (POM) were obtained: (1) free POM and (2) POM occluded in soil aggregates. Both POM fractions were analyzed by elemental analysis (C, N) and CPMAS ¹³C NMR spectroscopy. With increasing clay content the amount of organic carbon stored in the occluded POM fraction increased considerably, whereas the amounts of free POM were not related to the soil clay content. With increasing soil clay contents increasing proportions of O‐alkyl C and decreasing proportions of aryl C were found for both POM fractions. The occluded POM fraction showed a higher degree of degradation as indicated by lower amounts in O‐alkyl carbon. A lower degree of POM degradation was associated with higher clay contents. Higher soil clay contents promoted the conservation of POM with a low degree of alteration. This effect of soil texture was found to be highly significant when the aryl C : O‐alkyl C ratio was used as indicator for POM decomposition rather than the alkyl C : O‐alkyl C ratio.     

Soil organic matter quality and microbial activities in spruce swamp forests affected by drainage and water regime restoration

The effect of spruce swamp forest (SSF) drainage and water regime restoration on soil organic matter (SOM) quality and soil microbial heterotrophic activities was studied in pristine, drained and restored SSF in the Bohemian Forest, Czech Republic. Sequential chemical SOM fractionation using cold and hot water and hot acid was used to separate SOM fractions according to their mobility and potential lability/recalcitrance, and Fourier transform infrared spectra were used for SOM characterization. Soil physicochemical parameters and heterotrophic microbial activities were also determined. Drainage of SSF had significant long‐term effects (more than 50 yr) on plant communities and SOM quality. On drained sites, cover of sphagnum moss and sedge was much smaller than on pristine locations. A greater proportion of recalcitrant compounds and a smaller proportion of labile compounds were found in drained SSF as compared to pristine sites, which first led to an energy limitation and was followed by a decrease in microbial biomass and heterotrophic microbial activities (CO₂ production, methanogenesis and methanotrophy). Restoration resulted in slow progressive changes in the vegetation cover, including the spread of sphagnum mosses, retreat of mosses typical of drier conditions and increased sedge cover compared with drained SSF. Moreover, soil physicochemical parameters (pH and bulk density), hot‐water‐extractable C and methanotrophic activity tended to evolve towards the pristine SSF and seem to be good indicators of the restoration process. No other SOM fractions changed significantly after restoration. Thus, to change significantly overall SOM quality and most microbial heterotrophic activities following restoration, more than 7 yr are required.     

Free and intra‐aggregate organic matter as indicators of soil quality change in volcanic soils under contrasting crop rotations

Soil physical fractionation techniques may provide indicators of changing soil organic carbon (SOC) content; however, they have not been widely tested on volcanic soils (Andisols). In this study, we assessed two fractions as potential indicators in volcanic soils, using two sites in Chile converted from natural grassland to arable and mixed crop rotations, 8 and 16 yr previously. In the 8‐yr experiment, SOC had declined under all rotations, with smaller changes where the rotation included 3 or 5 yr of perennial pasture. Whereas the average SOC was only 76% of the level in the preceding natural grassland, the corresponding value after 16 yr for the second site was 98% (and 93% under continuous arable), probably reflecting its high allophane clay content. The fractionation procedure tested proved applicable to both Andisols, but the intra‐aggregate light fraction (IA‐SOM, isolated in sodium iodide solution at 1.80 g/cm³ after ultrasonic dispersion) accounted for a very small proportion of total SOC (<1%). We suggest that in Andisols, the free light fraction (FR‐SOM, isolated in sodium iodide at solution of the same density, but prior to ultrasonic dispersion) is stabilised to a greater extent than in nonvolcanic soils, and the intra‐aggregate fraction plays a more minor role as a pool of intermediate turnover. The relative value of each fraction needs to be confirmed through dynamic experiments, using more sites, and including situations where SOC content is initially low.     

不同土地利用下黑土密度分组中碳、氮的分配变化

陆地生态系统植被覆盖会影响有机质在土壤及其各组分中的分布,是控制土壤有机质储量的重要因子。通过对黑土有机碳、氮含量的估算,阐明土地利用变化对土壤有机碳库及轻组、重组有机碳、氮含量和结合态腐殖质有机碳含量的影响以及有机碳、氮的消长变化。结果表明:(1)不同土地利用方式下土壤0～20cm土层总有机碳(TOC)、全氮(TN)含量存在显著差异(p<0.05),自然土壤被破坏或转为农田,土壤总有机碳和全氮含量会持续下降,但农田化肥和有机肥配施后,土壤总有机碳和全氮含量显著增加;草地经过20年的植被恢复,土壤总有机碳含量显著高于农田化肥和无肥处理,有机碳库储量显著提高;(2)游离态轻组(Free-LF)和重组(HF)有机碳和氮含量与土壤总有机碳和全氮呈显著或极显著正相关,闭蓄态轻组(Occluded-LF)有机碳和氮在土壤中呈随机分布;(3)游离态轻组的C/N比高于全土和其他组分,易受微生物作用而降解,是土壤不稳定碳库的主要成分;(4)松结态胡敏酸碳含量与土壤总有机碳显著相关(p<0.05),土壤肥力随着胡敏酸的增加而提高。农田有机培肥不但能够提高有机质含量和土壤质量,也能显著提高农田生态系统抵押大气CO2的能力。     

Organic matter in size‐density fractions after 16–50 years of grass ley,cereal cropping and organic amendments

Size‐density fractionation, which was originally developed to examine short‐term decomposition of added material in sandy soil, was highly sensitive to medium‐ to long‐term changes in loam and clay soils. Materials from different size classes (>1 mm, 0.1–1 mm, 0.05–0.1 mm and <0.05 mm) were separated by density into light (ρ < 1.0 g cm⁻³), medium (1.0 < ρ < 1.85 g cm⁻³) and heavy (ρ > 1.85 g cm⁻³) fractions. In 16–18‐year cropping experiments the 0.1–1‐mm heavy fraction contained 17–19% of total carbon in ley soils compared with 7% in continuously cropped soils. Greater N‐mineralization rates after sieving of ley cropped soils could not be related to differences in C:N ratios of fractions, but this was assumed to be related to exposure of aggregate‐binding agents. In a 50‐year trial 40% of total soil carbon was contained in the 0.1–1‐mm heavy fraction in soil treated with sewage sludge compared with 7–9% in the fallow and ‘zero N’ treatments. Thus, the soils studied expressed an aggregate hierarchy dependent on organic carbon dynamics. The relative abundance of carbon in heavy organo‐mineral fractions >0.1 mm was inversely related to the relative abundance of C in black‐brown medium density material <0.1 mm, defined as uncomplexed particulate organic matter that was presumably released during ultrasonic disruption deterioration of finer (<0.1 mm) aggregated organo‐mineral particles. The size density fractionation allowed identification of materials of contrasting visual appearance, chemical qualities and, by inference, biological turnover times. However, they were found to be predominantly composite fractions and may be too complex to be represented by unique model pools.     

The contribution of various organic matter fractions to soil–water interactions and structural stability of an agriculturally cultivated soil

The presence and mutual interactions of soil organic matter (SOM) and clay particles are major factors determining soil structural stability. In the scope of agricultural management and environmental sustainability, it remains unclear how various mineral and organic matter (OM) fractions, OM–clay interactions and swelling processes in the interparticle space determine soil–water interactions and thus soil structural stability. To investigate this issue, we isolated the mineral and OM fractions of an agriculturally cultivated silty loam soil by soil density fractionation and assessed their hydration characteristics and effects on soil structural stability combining ¹H‐NMR relaxometry, soil rheology and single wet‐sieving of soil aggregates. The results showed that agricultural management practices, in particular compost and ploughing, as well as various OM–clay interactions significantly affected soil–water interactions and soil structural stability. On the one hand, ploughing reduced soil structural stability by promoting clay swelling as a result of disrupted soil structures and reduced SOM content. On the other hand, compost treatment and reduced tillage increased soil structural stability. In all cases, soil density fractionation showed that compost‐derived particulate organic matter (POM) and mineral‐associated organic matter (MAOM) restricted clay swelling and resulted in a highly porous and mechanically stable soil matrix. In particular, POM increased soil structural stability by acting as nucleus for soil aggregation and by restricting clay swelling via its presence as solid, granular interparticulate material. In contrast, MAOM seemed to restrict clay swelling via clay surface covering and the formation of viscous interparticulate hydrogel structures.     

A comparison of two methods for the isolation of free and occluded particulate organic matter

Various methods exist for the isolation of particulate organic matter (POM), one of the soil‐organic‐matter (SOM) fractions reacting most sensitive on land‐use or soil‐management changes. A combination of density separation and ultrasonic treatment allows to isolate two types of POM: (1) free POM and (2) POM occluded in soil aggregates. POM fractions are closely linked to their biochemical function for the formation and stabilization of aggregates, therefore methods using different aggregate sizes may result in different POM fractions isolated. We evaluated two physical fractionation procedures to reveal whether they yield different POM fractions with respect to amount and composition, using grassland and arable soils with sandy‐loam to sandy–clay‐loam texture and thus low macroaggregate stability. Method I used air‐dried aggregates of <2.0 mm size and a low‐energy sonication for aggregate disruption, method II used field‐moist aggregates <6.3 mm and a high‐energy–sonication procedure for aggregate disruption. POM fractions were analyzed by elemental analysis (C, N) and CPMAS ¹³C‐NMR spectroscopy. With both methods, about similar proportions of the SOM are isolated as free or occluded POM, respectively. The free‐ and occluded‐POM fractions obtained with method I are also rather similar in C and N concentration and composition as shown by ¹³C‐NMR spectroscopy. Method II isolates a free‐ and occluded‐POM fraction with significantly different C and N concentrations. NMR spectra revealed significant differences in the chemical composition of both fractions from method II, with the occluded POM having lower amounts of O‐alkyl C and higher amounts of aryl C and alkyl C than the free POM. Due to the use of larger, field‐moist aggregates with minimized sample pretreatment, two distinctly different POM fractions are isolated with method II, likely to be more closely linked to their biochemical function for the formation and stabilization of aggregates. High‐energy sonication as in method II also disrupts small microaggregates <63 µm and releases fine intraaggregate POM. This fraction seems to be a significant component of occluded POM, that allows a differentiation between free and occluded POM in sandy soils with significant microaggregation. It can be concluded, that microaggregation in arable soils with sandy texture is responsible for the storage of a more degraded occluded POM, that conversely supports the stabilization of fine microaggregates.     

Particle size fractionation of soil containing coal and combusted particles

Standard procedure for dispersing natural soils for particle size fractionation can be too aggressive for soil containing coal and other organic particles from coal industries. We have investigated ultrasonic dispersion for the latter in four soils differing in pedogenesis (Phaeozem, Podzol, reclaimed mine soils), carbon content (27.5–138.6 g kg^–1), clay content (80–153 g kg^–1) and sources of particles (airborne coal dust, combustion residues, lignite particles). As we found previously for natural soils, the ultrasonic energy needed for complete dispersion varies between 450 and 500 J ml^–1, but the resulting particle size distributions differ from those obtained by standard textural analysis. This is probably related to the different properties of native soil organic matter and coal and combusted particles. Coal and soot particles may partly resist oxidation with hydrogen peroxide, depending on material and particle size. The diameter of lignite particles, remaining after oxidation, is overestimated in sedimentation analysis by a factor of 1.66. Sand-sized lignite particles can be disrupted by ultrasonication and redistributed to finer particle size fractions. The ultrasonic dispersion and particle size fractionation procedure can be applied to soils containing coal and combusted particles, but caution is needed in interpreting the results if they contain large proportions of coal particles.     

Effect of air‐drying on phosphorus fractions in clay soil

Despite the publication of a number of papers dealing with the effect of drying on the soil labile P pool, less attention has been paid to the possible drying‐evoked changes in the more stable P pools. We applied Hedley's sequential fractionation procedure that aims at quantifying soil P reserves according to their decreasing plant availability to examine the effects of drying on soil P fractions in clayey soil samples of different cultivation history. To further investigate the contribution of organic matter disruption to the solubility of soil P, the P extracted in each fractionation step was divided into two size classes by filtering the suspension through a 0.2 μm membrane filter. There were no air‐drying‐induced changes in the total amount of P extracted in each fractionation step. However, air‐drying changed the distribution of water‐extractable P in size fractions; increase in the small‐sized P took place at the expense of large‐sized P. Air‐drying increased also small‐sized molybdate‐unreactive P (MUP) in the NaOH fraction giving evidence that drying‐induced alterations take place also in less labile P forms. The results revealed that air‐drying alters the extractability and distribution of P in various pools rather than the total amount of extracted P and that a large proportion of H₂O‐ and NaOH‐extractable large‐sized MUP may remain undetected if only filtered samples are analyzed.