Determination of the fate of 13C labelled maize and wheat exudates in an agricultural soil during a short-term incubation

A broader knowledge of the contribution of carbon (C) released by plant roots (exudates) to soil is a prerequisite for optimizing the management of organic matter in arable soils. This is the first study to show the contribution of constantly applied ¹³C‐labelled maize and wheat exudates to water extractable organic carbon (WEOC), microbial biomass‐C (MB‐C), and CO₂‐C evolution during a 25‐day incubation of agricultural soil material. The CO₂‐C evolution and respective δ¹³C values were measured daily. The WEOC and MB‐C contents were determined weekly and a newly developed method for determining δ¹³C values in soil extracts was applied. Around 36% of exudate‐C of both plants was recovered after the incubation, in the order WEOC < MB‐C < CO₂‐C for maize and MB‐C < WEOC < CO₂‐C for wheat. Around 64% of added exudate‐C was not retrieved with the methods used here. Our results suggest that great amounts of exudates became stabilized in non‐water extractable organic fractions. The amounts of MB‐C stayed relatively constant over time despite a continuous exudate‐C supply, which is the prerequisite for a growing microbial population. A lack of mineral nutrients might have limited microbial growth. The CO₂‐C mineralization rate declined during the incubation and this was probably caused by a shift in the microbial community structure. Consequently, incoming WEOC was left in the soil solution leading to rising WEOC amounts over time. In the exudate‐treated soil additional amounts of soil‐derived WEOC (up to 110 μg g⁻¹) and MB‐C (up to 60 μg g⁻¹) relative to the control were determined. We suggest therefore that positive priming effects (i.e. accelerated turnover of soil organic matter due to the addition of organic substrates) can be explained by exchange processes between charged, soluble C‐components and the soil matrix. As a result of this exchange, soil‐derived WEOC becomes available for mineralization.     

Aggregate-occluded black carbon in soil

The great stability of black carbon (BC) in soils may not be solely attributable to its refractory structure but also to poor accessibility when physically enveloped by soil particles. Our aim was to elucidate the intensity of physical entrapment of BC within soil aggregates. For this purpose, the A horizon of a forest, and of a grassland soil, and of three soils under tillage, were sampled at the experimental station Rotthalmünster, Germany. Black carbon was assessed in water‐stable aggregates and aggregate‐density fractions using benzene polycarboxylic acids as specific markers. The greatest BC concentrations made up 7.2% of organic carbon and were found in the < 53 μm fraction. The smallest BC concentrations occurred in the large macroaggregate fractions (> 2 mm). This pattern has been sustained even after tillage. The C‐normalized BC concentrations were significantly greater (P < 0.05) in the occluded particulate organic matter (OPOM) fractions than in the free particulate organic matter (FPOM) and the mineral fractions. This enrichment of BC compared with organic carbon in the OPOM fractions amounted to factors of 1.5–2.7. Hence, BC was embedded within microaggregates in preference to other organic carbon compounds. Only 2.5–3.5% of BC was located in the OPOM fraction < 1.6 g cm^?3, but 22–24% in the OPOM fraction with a density of 1.6–2.0 g cm^?3. This suggests that BC possibly acted as a binding agent or was selectively enriched during decomposition of protected SOM, or both. Physical inclusion, particularly within microaggregates, could therefore contribute to the long mean‐residence times of soil‐inherent BC.     

Location and chemical composition of stabilized organic carbon in topsoil and subsoil horizons of two acid forest soils

The ¹⁴C age of soil organic matter is known to increase with soil depth. Therefore, the aim of this study was to examine the stabilization of carbon compounds in the entire soil profile using particle size fractionation to distinguish SOM pools with different turnover rates. Samples were taken from a Dystric Cambisol and a Haplic Podzol under forest, which are representative soil types under humid climate conditions. The conceptual approach included the analyses of particle size fractions of all mineral soil horizons for elemental composition and chemical structure of the organic matter by ¹³C cross-polarization magic angle spinning nuclear magnetic resonance (CPMAS NMR) spectroscopy. The contribution of phenols and hydroxyalkanoic acids, which represent recalcitrant plant litter compounds, was analyzed after CuO oxidation.In the Dystric Cambisol, the highest carbon concentration as well as the highest percentage of total organic carbon are found in the <6.3 μm fractions of the B and C horizons. In the Haplic Podzol, carbon distribution among the particle size fractions of the Bh and Bvs horizons is influenced by the adsorption of dissolved organic matter. A relationship between the carbon enrichment in fractions <6.3 μm and the ¹⁴C activity of the bulk soil indicates that stabilization of SOM occurs in fine particle size fractions of both soils. ¹³C CPMAS NMR spectroscopy shows that a high concentration of alkyl carbon is present in the fine particle size fractions of the B horizons of the Dystric Cambisol. Decreasing contribution of O-alkyl and aromatic carbon with particle size as well as soil depth indicates that these compounds are not stabilized in the Dystric Cambisol. These results are in accordance with data obtained by wet chemical analyses showing that cutin/suberin-derived hydroxyalkanoic acids are preserved in the fine particle size fractions of the B horizons. The organic matter composition in particle size fractions of the top- and subsoil horizons of the Haplic Podzol shows that this soil is acting like a chromatographic system preserving insoluble alkyl carbon in the fine particle size fractions of the A horizon. Small molecules, most probably organic acids, dominate in the fine particle size fractions of the C horizons, where they are stabilized in clay-sized fractions most likely due to the interaction with the mineral phase. The characterization of lignin-derived phenols indicated, in accordance with the NMR measurements, that these compounds are not stabilized in the mineral soil horizons.     

Effect of soil pH on the chemical composition of organic matter in physically separated soil fractions in two broadleaf woodland sites at Rothamsted,UK

Although acid soils are common in forest ecosystems, and there is documented evidence of pH influencing transformations of organic matter in soil, there are surprisingly few studies on the influence of soil pH on the chemical structure of physically fractionated soil organic matter (SOM). The aim of this study was to characterize the influence of pH on the chemical and physical processes involved in SOM stabilization. Forest soils of different pH (4.4 and 7.8) sampled from two long‐term experiments at Rothamsted Research (UK) were physically fractionated. The free light fraction (FLF), the intra‐aggregate light fraction and the fine silt and clay (S + C, <25 µm) were characterized using elemental, isotopic (δ¹³C), thermogravimetric, differential thermal, diffuse reflectance infrared Fourier transform spectroscopy and high‐resolution magic angle spinning ¹H nuclear magnetic resonance analyses. The quantitative distribution of carbon (C) between SOM fractions differed between the two soils. Carbon contents in the light fractions from the acid soil were significantly greater than in those of the alkaline soil. In contrast, in S + C fractions, C content was greater in the alkaline soil. FLF from the acid soil was characterized by a greater C:N ratio, smaller δ¹³C and greater content of thermo‐labile compounds compared with FLF from the alkaline soil. In contrast, there was only a weak effect of soil pH on the chemical composition of the organic matter in S + C fractions. Irrespective of soil pH, these latter fractions contained mainly aliphatic compounds such as carbohydrates, carboxylic acid, amide and peptide derivates. This suggested that physical mechanisms, involving the interactions between SOM and mineral surfaces, are of greater importance than the presence of chemically recalcitrant species in protecting SOM associated with the finest soil fractions.     

Stable carbon isotope ratios of water-extractable organic carbon affected by application of rice straw and rice straw compost during a long-term rice experiment in Yamagata,Japan

ABSTRACT

Hot-water- and water-extractable organic matter were obtained from soil samples collected from a rice paddy 31 years after the start of a long-term rice experiment in Yamagata, Japan. Specifically, hot-water-extractable organic carbon and nitrogen (HWEOC and HWEON) were obtained by extraction at 80°C for 16 h, and water-extractable organic carbon and nitrogen (WEOC and WEON) were obtained by extraction at room temperature. The soil samples were collected from surface (0–15 cm) and subsurface (15–25 cm) layers of five plots that had been treated with inorganic fertilizers alone or with inorganic fertilizers plus organic matter, as follows: PK, NPK, NPK plus rice straw (RS), NPK plus rice straw compost (CM1), and NPK plus a high dose of rice straw compost (CM3). The soil/water ratio was 1:10 for both extraction temperatures. We found that the organic carbon and total nitrogen contents of the bulk soils were highly correlated with the extractable organic carbon and nitrogen contents regardless of extraction temperature, and the extractable organic carbon and nitrogen contents were higher in the plots that were treated with inorganic fertilizers plus organic matter than in the PK and NPK plots. The HWEOC and WEOC δ¹³C values ranged from ?28.2% to ?26.4% and were similar to the values for the applied rice straw and rice straw compost. There were no correlations between the HWEOC or WEOC δ¹³C values and the amounts of HWEOC or WEOC. The δ¹³C values of the bulk soils ranged from ?25.7% to ?23.2% and were lower for the RS and CM plots than for the PK and NPK plots. These results indicate that HWEOC and WEOC originated mainly from rice plants and the applied organic matter rather than from the indigenous soil organic matter. The significant positive correlations between the amounts of HWEOC and HWEON and the amount of available nitrogen (P < 0.001) imply that extractable organic matter can be used as an index for soil fertility in this long-term experiment. We concluded that the applied organic matter decomposed more rapidly than the indigenous soil organic matter and affected WEOC δ¹³C values and amounts.     

Changes in soil organic matter composition are associated with forest encroachment into grassland with long-term fire history

This study investigates if Araucaria forest (C₃ metabolism) expansion on frequently burnt grassland (C₄ metabolism) in the southern Brazilian highland is linked to the chemical composition of soil organic matter (SOM) in non‐allophanic Andosols. We used the ¹³C/¹²C isotopic signature to group heavy organo‐mineral fractions according to source vegetation and ¹³C NMR spectroscopy, lignin analyses (CuO oxidation) and measurement of soil colour lightness to characterize their chemical compositions. Large proportions of aromatic carbon (C) combined with small contents of lignin‐derived phenols in the heavy fractions of grassland soils and grass‐derived lower horizons of Araucaria forest soils indicate the presence of charred grass residues in SOM. The contribution of this material may have led to the unusual increase in C/N ratios with depth in burnt grassland soils and to the differentiation of C₃‐ and C₄‐derived SOM, because heavy fractions from unburnt Araucaria forest and shrubland soils have smaller proportions of aromatic C, smaller C/N ratios and are paler compared with those with C₄ signatures. We found that lignins are not applicable as biomarkers for plant origin in these soils with small contents of strongly degraded and modified lignins as the plant‐specific lignin patterns are absent in heavy fractions. In contrast, the characteristic contents of alkyl C and O/N‐alkyl C of C₃ trees or shrubs and C₄ grasses are reflected in the heavy fractions. They show consistent changes of the (alkyl C)/(O/N‐alkyl C) ratio and the ¹³C/¹²C isotopic signature with soil depth, indicating their association with C₄ and C₃ vegetation origin. This study demonstrates that soils may preserve organic matter components from earlier vegetation and land‐use, indicating that the knowledge of past vegetation covers is necessary to interpret SOM composition.     

Predicting soil organic matter stability in agricultural fields through carbon and nitrogen stable isotopes

In order to evaluate the sustainability and efficiency of soil carbon sequestration measures and the impact of different management and environmental factors, information on soil organic matter (SOM) stability and mean residence time (MRT) is required. However, this information on SOM stability and MRT is expensive to determine via radiocarbon dating, precluding a wide spread use of stability measurements in soil science. In this paper, we test an alternative method, first developed by Conen et al. (2008) for undisturbed Alpine grassland systems, using C and N stable isotope ratios in more frequently disturbed agricultural soils. Since only information on carbon and nitrogen concentrations and their stable isotope ratios is required, it is possible to estimate the SOM stability at greatly reduced costs compared to radiocarbon dating. Using four different experimental sites located in various climates and soil types, this research proved the effectiveness of using the C/N ratio and δ¹⁵N signature to determine the stability of mOM (mineral associated organic matter) relative to POM (particulate organic matter) in an intensively managed agro-ecological setting. Combining this approach with δ¹³C measurements allowed discriminating between different management (grassland vs cropland) and land use (till vs no till) systems. With increasing depth the stability of mOM relative to POM increases, but less so under tillage compared to no-till practises. Applying this approach to investigate SOM stability in different soil aggregate fractions, it corroborates the aggregate hierarchy theory as proposed by Six et al. (2004) and Segoli et al. (2013). The organic matter in the occluded micro-aggregate and silt & clay fractions is less degraded than the SOM in the free micro-aggregate and silt & clay fractions. The stable isotope approach can be particularly useful for soils with a history of burning and thus containing old charcoal particles, preventing the use of ¹⁴C to determine the SOM stability.     

The isotopic composition of soil organic carbon on a north–south transect in western Canada

The minor isotopes of carbon (¹³C and ¹⁴C) are widely used as tracers in studies of the global carbon cycle. We present carbon‐isotope data for the 0–5 cm layer of soil on a transect from 49.6°N to 68°N, from mature forest and tundra ecosystems in the boreal‐arctic zone of interior western Canada. Soil organic carbon in the < 2000 μm fraction of the soil decreases from 3.14 kg m^?2 in the south to 1.31 kg m^?2 in the north. The ¹⁴C activity of the organic carbon decreases as latitude increases from 118.9 to 100.7 per cent modern carbon (pMC). In addition, the ¹⁴C activities of organic carbon in the particle‐size fractions of each sample decrease as particle size decreases. These results suggest that organic carbon in the 0–5 cm layer of these soils transfers from standing biomass into the coarsest size fractions of the soil and is then degraded over time, with the residue progressively transferred into the more resistant finer particle sizes. We calculate residence times for the coarsest size fractions of 21 years in the south to 71 years in the north. Residence times for the fine size fractions (< 63 μm) are considerably longer, ranging from 90 years in the south to 960 years in the north. The δ¹³C of the organic carbon decreases from ?26.8 ± 0.3‰ in soil under forest in the south to ?26.2 ± 0.1‰ for tundra sites in the north. At all sites there is an increase in δ¹³C with decreasing particle size of 0.7–1.6‰. These changes in δ¹³C are due to the presence of ‘old’ carbon in equilibrium with an atmosphere richer in ¹³C, and to the effects of microbial degradation.     

The sensitivity of water extractable soil organic carbon fractions to land use in three soil types

Soil organic carbon (SOC) has a high impact on the sustainability of ecosystems, global environmental processes, soil quality and agriculture. Long-term tillage usually leads to SOC depletion. The purpose of this study was to determine the impact of different land uses on water extractable organic carbon (WEOC) fractions and to evaluate the interaction between the WEOC fractions and other soil properties. Using an extraction procedure at 20°C and 80°C, two fractions were obtained: a cold water extractable organic carbon (CWEOC) and a hot water extractable organic carbon (HWEOC). The results suggest that there is a significant impact from different land uses on WEOC. A lower relative contribution of WEOC in SOC and a lower concentration of labile WEOC fractions are contained in arable soil compared to forestlands. Chernozem soil was characterized by a lower relative contribution of WEOC to the SOC and thus higher SOC stability in contrast to Solonetz and Vertisol soils. Both CWEOC and HWEOC are highly associated with SOC in the silt and clay fraction (<53 µm) and were slightly associated with SOC in the macroaggregate classes. The WEOC fractions were highly and positively correlated with the SOC and mean weight diameter.     

Soil organic carbon stocks,distribution, and composition affected by historic land use changes on adjacent sites

Historic alterations in land use from forest to grassland and cropland to forest were used to determine impacts on carbon (C) stocks and distribution and soil organic matter (SOM) characteristics on adjacent Cambisols in Eastern Germany. We investigated a continuous Norway spruce forest (F-F), a former cropland afforested in 1930 (C-F), and a grassland deforested in 1953 (F-G). For C and N stocks, we sampled the A and B horizons of nine soil pits per site. Additionally, we separated SOM fractions of A and B horizons by physical means from one central soil pit per pedon. To unravel differences of SOM composition, we analyzed SOM fractions by ¹³C-CPMAS NMR spectroscopy and radiocarbon analysis. For the mineral soils, differences in total C stocks between the sites were low (F-F = 8.3 kg m⁻²; C-F = 7.3 kg m⁻²; F-G = 8.2 kg m⁻²). Larger total C stocks (+25%) were found under continuous forest compared with grassland, due to the C stored within the organic horizons. Due to a faster turnover, the contents of free particulate organic matter (POM) were lower under grassland. High alkyl C/O/N-alkyl C ratios of free POM fractions indicated higher decomposition stages under forest (1.16) in relation to former cropland (0.48) and grassland (0.33). Historic management, such as burning of tree residues, was still identifiable in the subsoils by the composition and ¹⁴C activity of occluded POM fractions. The high potential of longer lasting C sequestration within fractions of slower turnover was indicated by the larger amounts of claybound C per square meter found under continuous forest in contrast to grassland.     

Black carbon in the zonal steppe soils of Russia

Black Mollisols are typically rich in charred organic matter, however, little is known about the zonal distribution of black C (BC) in steppe soils. In this study, we used benzene polycarboxylic acids (BPCA) as specific markers for BC in particle‐size fractions of depth profiles in several zonal soils (Greyzem, Phaeozem, Chernozem, Kastanozem) of the Russian steppe. In addition, liquid‐state ¹³C‐NMR spectra were obtained on the alkaline‐soluble soil organic matter (SOM). The results showed that both the content and depth distribution of BC varies in the different soil types; the concentration of BC in the bulk top soils being closely related to the aromaticity of the SOM (r² = 0.98 for the native topsoils, 0.83 for top‐ and subsurface soils). Especially the Chernozems were rich in aromatic SOM, which partly contained more than 17% BC of total C, most of which being allocated in the mineral fractions. Long‐term arable cropping did not reduce the BC contents of the surface soil, though it did promote the enrichment of BC in the silt fractions. The same shift was detected as soil depth increased. We conclude that BC is not fully inert in these soils, but apparently can be preserved in the silt as decomposition of SOM increased, i.e., it accumulates exactly in that fraction, which has been formerly assigned to contain old, aromatic C.     

Stabilization of carbon from maize in a sandy soil in a long-term experiment

The sequestration of carbon (C) in soil is not completely understood, and quantitative information about the amounts of organic carbon in the various fractions and their rates of turnover could improve understanding. We aimed (i) to quantify the amounts of C derived from maize at various depths in the soil in a long‐term field experiment with and without fertilization using ¹³C/¹²C analysis, (ii) to model changes in the organic C, and (iii) to compare measured and modelled pools of C. The organic C derived from the maize was measured in soil samples collected to a depth of 65 cm from four plots, two of which had been under continuous maize and two under continuous rye during long‐term field experiments with NPK and without fertilization. The fractionation procedures included particle‐size fractionation and extractions in water and in pyrophosphate solution. We used the Rothamsted Carbon Model to model the dynamics of the carbon from ¹³C data. The amounts of C derived from maize in the Ap horizon after 39 years of continuous maize cropping were 9.5% of the total organic C (where unfertilized) and 14.0% where NPK had been applied. Fertilization did not affect the residence time of carbon in the soil. The amounts of C derived from maize in water extracts were 21% of the total organic C (where unfertilized) and 22% where NPK had been applied. The extracts that were soluble in pyrophosphate and insoluble in acid were depleted in C from maize (the amounts were 5% and 7% of the total organic C, respectively). The results of the ¹³C natural abundance technique were used to model the dynamics of the organic C. Both the total organic C and the C derived from maize in the particle‐size fraction 0–63 μm agreed well with the total and maize‐derived sums of the model pools ‘inert organic matter’, ‘humified organic matter’ and ‘microbial biomass’. The model suggested that 64% (unfertilized) or 53% (NPK) of the organic C in the Ap horizon were inert. Only one of three published equations to determine the size of the inert pool agreed well with these model results.     

The influence of soil processes on carbon isotope distribution and turnover in the British uplands

Understanding the natural variation of carbon within the soil, and between soil types, is crucial to improve predictive models of carbon cycling in high and mid-latitude ecosystems in response to global warming. We measured the carbon isotope distributions (¹²C, ¹³C and ¹⁴C) in soil organic matter (SOM) from Podzols, Brown Podzolic soils and Stagnohumic Gleysols from the British uplands, which were then compared with the total amounts and turnover of carbon in these soils. We did so by sampling at 2-cm intervals down six profiles of each soil type. The average amount of carbon stored in the top 28 cm of the Stagnohumic Gleysols is twice that of the other two soils. The ¹³C content and ¹⁴C age show a general increase with depth in all soils, and there is also a significant correlation between isotopic variation and the main pedogenic features. The latter suggests that soil-forming processes are significant in determining the carbon isotope signatures retained in SOM. Organic matter formed since 1960 is not found below 5 cm in any of the soils. Evidently organic detritus in the surface layers (LF and Oh) is rapidly mineralized. This accords with our modelled net annual C fluxes which show that more than 80% of the CO₂ emanating from these soils is derived from the top 5 cm of each profile. Although these soils contain much carbon, they do not appear to assimilate and retain SOM rapidly. The mean residence time of most of their carbon is in the 2–50 years range, so the soils are fairly ineffective sinks for excess CO₂ in the atmosphere. Under the predicted future ‘greenhouse’ climate, likely to favour more rapid microbial decomposition of organic materials, these soils are a potential source of CO₂ and are therefore likely to accelerate global warming.     

How are soil use and management reflected by soil organic matter characteristics: a spectroscopic approach

We studied the quantitative and qualitative changes of soil organic matter (SOM) due to different land uses (arable versus grassland) and treatments (organic manure and mineral fertilizer) within an agricultural crop rotation in a long‐term field experiment, conducted since 1956 at Ultuna, Sweden, on a Eutric Cambisol. The organic carbon (OC) content of the grassland plot was 1.8 times greater than that of the similarly fertilized Ca(NO₃)₂ treated cropped plots. The comparison of two dispersion techniques (a low‐energy sonication and a chemical dispersion which yield inherent soil aggregates) showed that increasing OC contents of the silt‐sized fractions were not matched by a linear increase of silt‐sized aggregates. This indicated saturation of the aggregates with OC and a limited capacity of particles to protect OC physically. Thermogravimetric analyses suggested an increase of free organic matter with increasing OC contents. Transmission FT‐IR spectroscopy showed relative enrichment of carboxylic, aromatic, CH and NH groups in plots with increasing OC contents. The silt‐sized fractions contained the largest SOM pool and, as revealed by ¹³C NMR spectroscopy, were qualitatively more influenced by the plant residue versus manure input than the clay fractions. Alkyl and O‐alkyl C in the silt‐sized fractions amounted to 57.4% of organic carbon in the animal manure treated plots and 50–53% in the other treatments.     

Land-use related organic matter dynamics in North Cameroon soils assessed by 13C analysis of soil organic matter fractions

Topsoil samples from cultivated and adjacent non‐cultivated fields on three major agricultural soils in North Cameroon were fractionated into particle‐size fractions that were analysed subsequently for their C and ¹³C contents. The aim was to obtain further insight into the dynamics of soil organic matter (SOM) in relation to land use in Cameroon. Since organic carbon contents of the fractions were often very small, samples and analyses were extensively replicated to obtain robust statistical estimates of observed differences. For each soil type, differences in δ¹³C values between fields could be related to changes in the input and decomposition of organic matter arising from soil type, land management and, for example, the nature and abundance of weeds. Turnover of organic matter appeared to be fastest in the sand fraction, which is in line with results from earlier studies. In the finer fractions, clear differences in reaction to changes in input and decomposition were observed, that seem to be linked to differences in clay mineralogy. The results illustrate that SOM in the various fractions is much less stable and more strongly affected by changes in land use than might be assumed on the basis of changes in total SOM contents alone. At the same time, they demonstrate the relevance of ¹³C isotope analyses of SOM for studies on the impact of land use on these savannah soils with little SOM that are highly susceptible to degradation.     

森林生态系统中土壤有机质组分的固态13C核磁共振波谱表征

Forest management practices such as prescribed burning and thinning in forest ecosystems may alter the properties of soil organic matter (SOM).In this study,surface soils from field plots in the Bankhead National Forest,Alabama,USA,were used to investigate possible SOM transformations induced by thinning and burning.Elemental analysis and solid-state 13C cross polarization magic angle spinning nuclear magnetic resonance (13C CPMAS NMR) spectroscopy were used to characterize SOM fractions in whole soils,humic substances,and density fractions.Our data revealed that the changes in SOM fractions due to the repeated burning carried out in the forest ecosystem studied were involved mainly with alkyl C,O-alkyl C,and carbohydrate functional groups,implying that most prominent reactions that occurred involved dehydrogenation,de-oxygenation,and decarboxylation.In addition,burning and thinning might have also affected the distribution and composition of free and occluded particulate SOM fractions.The limited structural changes in SOM fractions suggested that low-intensity prescribed fire in the forest ecosystem studied will not create major structural changes in SOM fractions.     

Comparison of chemical fractionation methods for isolating stable soil organic carbon pools

Stable soil organic matter (SOM) is important for long‐term sequestration of soil organic carbon (SOC), but the usefulness of different fractionation methods to isolate stable SOM is open to question. We assessed the suitability of five chemical fractionation methods (stepwise hydrolysis, treatment with H₂O₂, Na₂S₂O₈, NaOCl, and demineralization of the NaOCl‐resistant fraction (NaOCl + HF)) to isolate stable SOM from soil samples of a loamy sand and a silty loam under different land use regimes (grassland, forest and arable crops). The apparent C turnover time and mean age of SOC before and after fractionation was determined by ¹³C and ¹⁴C analysis. Particulate organic matter was removed by density fractionation before soils were exposed to chemical fractionation. All chemical treatments induced large SOC losses of 62–95% of the mineral‐associated SOC fraction. The amounts of H₂O₂‐ and Na₂S₂O₈‐resistant SOC were independent from land use, while those of NaOCl‐ (NaOCl + HF)‐ and hydrolysis‐resistant SOC were not. All chemical treatments caused a preferential removal of young, maize‐derived SOC, with Na₂S₂O₈ and H₂O₂ being most efficient. The mean ¹⁴C age of SOC was 1000–10000 years greater after chemical fractionation than that of the initial, mineral‐associated SOC and mean ¹⁴C ages increased in the order: NaOCl < NaOCl + HF ≤ stepwise hydrolysis ≪ H₂O₂≈ Na₂S₂O₈. None of the methods appeared generally suitable for the determination of the inert organic matter pool of the Rothamsted Carbon Model. Nonetheless, our results indicate that all methods are able to isolate an older, more stable SOC fraction, but treatments with H₂O₂ and Na₂S₂O₈ were the most efficient ones in isolating stable SOM.     

Soil organic matter in soil depth profiles: Distinct carbon preferences of microbial groups during carbon transformation

This study investigates how carbon sources of soil microbial communities vary with soil depth. Microbial phospholipid fatty acids (PLFA) were extracted from 0–20, 20–40 and 40–60 cm depth intervals from agricultural soils and analysed for their stable carbon isotopes (δ¹³C values). The soils had been subjected to a vegetation change from C3 (δ¹³C≈?29.3‰) to C4 plants (δ¹³C≈?12.5‰) 40 years previously, which allowed us to trace the carbon flow from plant-derived input (litter, roots, and root exudates) into microbial PLFA. While bulk soil organic matter (SOM) reflected ≈12% of the C4-derived carbon in top soil (0–20 cm) and 3% in deeper soil (40–60 cm), the PLFA had a much higher contribution of C4 carbon of about 64% in 0–20 cm and 34% in 40–60 cm. This implies a much faster turnover time of carbon in the microbial biomass compared to bulk SOM. The isotopic signature of bulk SOM and PLFA from C4 cultivated soil decreases with increasing soil depth (?23.7‰ to ?25.0‰ for bulk SOM and ?18.3‰ to ?23.3‰ for PLFA), which demonstrates decreasing influence of the isotopic signature of the new C4 vegetation with soil depth. In terms of soil microbial carbon sources this clearly shows a high percentage of C4 labelled and thus young plant carbon as microbial carbon source in topsoils. With increasing soil depth this percentage decreases and SOM is increasingly used as microbial carbon source. Among all PLFA that were associated to different microbial groups it could be observed that (a) depended on availability, Gram-negative and Gram-positive bacteria prefer plant-derived carbon as carbon source, however, (b) Gram-positive bacteria use more SOM-derived carbon sources while Gram-negative bacteria use more plant biomass. This tendency was observed in all three-depth intervals. However, our results also show that microorganisms maintain their preferred carbon sources independent on soil depth with an isotopic shift of 3–4‰ from 0–20 to 40–60 cm soil depth.     

Disruption of soil aggregates by varied amounts of ultrasonic energy in fractionation of organic matter of a clay Latosol: carbon,nitrogen and δ13C distribution in particle‐size fractions

Ultrasonic energy has been widely used to disrupt soil aggregates before fractionating soil physically when studying soil organic matter (SOM). Nevertheless, there is no consensus about the optimum energy desirable to disrupt the soil. We therefore aimed (i) to quantify the effect of varied ultrasonic energies on the recovery of each particle‐size fraction and their C, N and δ¹³C distribution, and (ii) to determine an ideal energy to fractionate SOM of a specific soil. Our results show that the 2000–100 μm particle‐size fraction was composed mainly of unstable aggregates and the 100–2 μm fraction of stable aggregates. Energies of 260–275 J ml^?1 were sufficient to disrupt most of the unstable aggregates and leave stable aggregates. The use of this threshold energy combined with particle‐size fractionation was not satisfactory for all purposes, since litter‐like material and relatively recalcitrant organic carbon present in stable aggregates > 100 μm were recovered in the same pool. An ultrasonic energy of 825 J ml^?1 was not sufficient to stabilize the redistribution of soil mass and organic matter among particle‐size fractions, but at energies exceeding 260–275 J ml^?1 relatively stable aggregates would fall apart and cause a mixture of carbon with varied nature in the clay fraction.     

The dynamics of organic matter in rock fragments in soil investigated by 14C dating and measurements of 13C

Rock fragments in soil can contain significant amounts of organic carbon. We investigated the nature and dynamics of organic matter in rock fragments in the upper horizons of a forest soil derived from sandstone and compared them with the fine earth fraction (<2 mm). The organic C content and its distribution among humic, humin and non‐humic fractions, as well as the isotopic signatures (Δ¹⁴C and δ¹³C) of organic carbon and of CO₂ produced during incubation of samples, all show that altered rock fragments contain a dynamic component of the carbon cycle. Rock fragments, especially the highly altered ones, contributed 4.5% to the total organic C content in the soil. The bulk organic matter in both fine earth and highly altered rock fragments in the A1 horizon contained significant amounts of recent C (bomb ¹⁴C), indicating that most of this C is cycled quickly in both fractions. In the A horizons, the mean residence times of humic substances from highly altered rock fragments were shorter than those of the humic substances isolated in the fine earth. Values of Δ¹⁴C of the CO₂ produced during basal respiration confirmed the heterogeneity, complexity and dynamic nature of the organic matter of these rock fragments. The weak ¹⁴C signatures of humic substances from the slightly altered rock fragments confirmed the importance of weathering in establishing and improving the interactions between rock fragments and surrounding soil. The progressive enrichment in ¹³C from components with high‐¹⁴C (more recent) to low‐¹⁴C (older) indicated that biological activity occurred in both the fine and the coarse fractions. Hence the microflora utilizes energy sources contained in all the soil compartments, and rock fragments are chemically and biologically active in soil, where they form a continuum with the fine earth.