A novel GC/MS technique to assess N and C incorporation into soil amino sugars

Amino sugars have been used as biomarker to indicate microorganism contribution to soil organic matter turnover and sequestration. However, there is no direct gas chromatograph mass spectrometry (GC/MS) approach to assess microbial synthesis of amino sugars in soil. We developed a novel method which combines laboratory incubation of substrate containing ¹⁵N or ¹³C and a GC/MS technique to trace ¹⁵N or ¹³C isotope changes in three amino sugars, glucosamine, galactosamine, and muramic acid. Sample preparation followed the procedure of Zhang and Amelung (1996) [Zhang, X., Amelung, W., 1996. Gas chromatographic determination of muramic acid, glucosamine, galactosamine, and mannosamine in soils. Soil Biology and Biochemistry 28, 1201-1206.]. The GC/MS determination was conducted using a full scan mode with both electronic ionization (EI) and chemical ionization (CI) sources. The CI source was suitable for all of the three amino sugars, while the EI source was not applicable to muramic acid due to its low sensitivity in the determination as well as low concentration of muramic acid in soil. The enrichment of ¹⁵N or ¹³C in amino sugars during incubation was estimated by calculating the atom percentage excess (APE). ¹⁵N incorporation was evaluated according to fragment (F) abundance ratio of mass F+1 to F, whilst ¹³C incorporation was estimated according to the ratio of mass F+n to F (n is skeleton carbon number in the fragment). This novel method was assessed by using two soil samples (a Kandiudult and a Udoll) incubated with either ¹⁵N-amonium or U-¹³C-glucose. The results indicate that the GC/MS determination is reproducible, thus this technique is useful in detecting the microbial synthesis of amino sugars in soil, and especially it should be possible when looking at the position or how much labeled carbon and nitrogen atoms have been incorporated.     

Natural abundance of 15N in nitrate,ureides, and amino acids from plant tissues

The natural ¹⁵N abundances (δ¹⁵N values) were measured for nitrate and free and bound amino acids from the leaves of field-grown spinach (Spinacia oleracea L.) and komatsuna (Brassica campestris L.), as well as ureides and free and bound amino acids in the leaves and roots of hydroponically grown soybean (Glycine max L.) totally depending on dinitrogen. Nitrate from the spinach and komatsuna leaves and ureides from leaves and roots of soybean showed higher δ¹⁵N values than the total tissue N and N in free or bound amino acid fractions. The δ¹⁵N values of individual free and bound amino acids, determined by GC/C/MS using their acetylpropyl derivatives, were similar in leaf tissues except for proline but varied in soybean root tissues. The order of ¹⁵N enrichment was similar in the four samples: aspartic acid > glutamic acid > threonine, proline, valine > glycine + alanine +serine, γ-amino butyric acid, and phenylalanine.     

C and N natural abundance of the soil microbial biomass

Stable isotope analysis is a powerful tool in the study of soil organic matter formation. It is often observed that more decomposed soil organic matter is ¹³C, and especially ¹⁵N-enriched relative to fresh litter and recent organic matter. We investigated whether this shift in isotope composition relates to the isotope composition of the microbial biomass, an important source for soil organic matter. We developed a new approach to determine the natural abundance C and N isotope composition of the microbial biomass across a broad range of soil types, vegetation, and climates. We found consistently that the soil microbial biomass was ¹⁵N-enriched relative to the total (3.2 ‰) and extractable N pools (3.7 ‰), and ¹³C-enriched relative to the extractable C pool (2.5 ‰). The microbial biomass was also ¹³C-enriched relative to total C for soils that exhibited a C3-plant signature (1.6 ‰), but ¹³C-depleted for soils with a C4 signature (−1.1 ‰). The latter was probably associated with an increase of annual C3 forbs in C4 grasslands after an extreme drought. These findings are in agreement with the proposed contribution of microbial products to the stabilized soil organic matter and may help explain the shift in isotope composition during soil organic matter formation.     

土壤C/N对苹果植株生长及氮素利用的影响

土壤C/N是土壤氮素循环的重要影响因素。本研究以2年生"富士"/平邑甜茶为试验材料, 应用¹⁵N示踪技术研究了不同土壤C/N[6.21(CK)、10、15、20、25、30、35和40]对苹果植株生长及氮素利用和损失的影响。结果表明: 随着土壤C/N比值的逐渐增大, 苹果新梢长度和植株鲜重均呈先升高后降低的变化趋势, C/N=15、20和25的3个处理苹果新梢长度和植株鲜重最大, 三者间无显著差异, 但均显著高于其他处理。不同C/N处理间植株15N利用率存在差异, 土壤C/N=25时, 植株¹⁵N利用率最大, 为22.87%, 与C/N=20的处理间无显著差异, 但两者均显著高于其他处理; 土壤C/N=40时, 植株¹⁵N利用率最低, 仅为15.43%, 低于CK处理的16.65%。土壤C/N处于15~25时, 植株吸收的氮素来自于肥料氮的比例较高; 而土壤C/N较低(<15)或太高(>25)时, 植株吸收的氮素来自于土壤氮的比例较高。土壤氮素残留量随土壤C/N的增大逐渐增加, C/N=40处理的土壤氮素残留量是CK的1.32倍。随着土壤C/N比值的逐渐增大, 肥料氮损失量呈先减少后增加的变化趋势, 以C/N=25时最少, 仅为施氮量的49.87%, 而对照最大, 为61.54%。因此, 综合土壤C/N对苹果植株生长及氮素平衡状况来看, 土壤C/N为15~25时, 能促进植株的生长发育, 降低氮肥损失, 提高肥料利用率。     

Dissipation of bacterially derived C and N through the meso- and macrofauna of a grassland soil

Feeding relationships between organisms may be determined by observations of behaviour in manipulative experiments or, as in more recent times, by the use of stable isotope labelling to trace the passage of ¹³C and ¹⁵N through food webs. Here we introduce living bacteria, labelled with both ¹³C and ¹⁵N into intact soil cores to understand further the movement of bacterially sourced C and N into the meso- and macrofauna of a grassland soil. We found that these groups showed a range of isotope levels which relate to their feeding strategies. Some had no label (e.g. dipterous larvae), whilst others were highly labelled which may indicate a preference for the added bacteria. This latter group included Collembola, generally perceived as being predominantly fungal feeders. This work describes a novel technique which has the potential to provide critical information about the dissipation of bacterially derived C and N through the soil food web.     

Plant uptake of dual-labeled organic N biased by inorganic C uptake: Results of a triple labeling study

Direct plant uptake of organic nitrogen (N) is often studied using the dual-labeling approach (¹⁵N + ¹³C or ¹⁵N + ¹⁴C). However, the method might be hampered by uptake of labeled inorganic carbon (C) produced by mineralization of labeled organic compounds. Here we report the results from a triple labeling experiment (¹⁵N + ¹³C + ¹⁴C) investigating whether root uptake of labeled inorganic C can bias the results obtained in studies of organic N uptake using dual-labeled amino acids (¹⁵N, ¹³C). In a rhizosphere tube experiment we investigated ¹³C and ¹⁴C uptake by maize either supplied with labeled glycine or , but found no differences in uptake rates between these C-sources. The uptake of inorganic C to the shoot tissue was higher for maize grown in full light compared to shading, which indicates a passive uptake of inorganic C with water. We conclude that uptake of inorganic C produced by mineralization of amino acids can significantly bias the interpretations of organic N uptake studies using dual-labeling.     

Compartmentalization of the soil animal food web as indicated by dual analysis of stable isotope ratios (N/N and C/C)

The soil animal food web has become a focus of recent ecological research but trophic relationships still remain enigmatic for many taxa. Analysis of stable isotope ratios of N and C provides a powerful tool for disentangling food web structure. In this study, animals, roots, soil and litter material from a temperate deciduous forest were analysed. The combined measurement of δ¹⁵N and δ¹³C provided insights into the compartmentalization of the soil animal food web. Leaf litter feeders were separated from animals relying mainly on recent belowground carbon resources and from animals feeding on older carbon. The trophic pathway of leaf litter-feeding species appears to be a dead end, presumably because leaf litter feeders (mainly diplopods and oribatid mites) are unavailable to predators due to large size and/or strong sclerotization. Endogeic earthworms that rely on older carbon also appear to exist in predator-free space. The data suggest that the largest trophic compartment constitutes of ectomycorrhizal feeders and their predators. Additionally, there is a smaller trophic compartment consisting of predators likely feeding on enchytraeids and potentially nematodes.     

Determination of 15N abundance of amino acids in soil hydrolysates

¹⁵N abundance of amino acids in soil hydrolysates was determined by emission spectroscopic method.     

Interference by amino acids during the determination of N ammonium in soil

In the study of terrestrial N cycling, NH₄⁺ concentration and ¹⁵N enrichment are routinely determined by colorimetric continuous flow analysis and microdiffusion methods. Amino acids can interfere in these determinations; consequently the aim of the present study was to evaluate the significance of the interference. Glycine and glutamine are key amino acids in soil and were therefore used as ‘models’. Both glycine and glutamine interfered during continuous flow analysis, whereas interference during microdiffusion was of little importance. The effects of interference can be significant, e.g. estimates of gross mineralisation rate were reduced up to 33%, where we allowed for amino acid interference during determination of NH₄⁺ concentration. The potential influence of amino acid interference emphasises that development of continuous flow analysis to increase NH₄⁺ specificity is needed.     

Organic C and N stabilization in a forest soil: Evidence from sequential density fractionation

In mineral soil, organic matter (OM) accumulates mainly on and around surfaces of silt- and clay-size particles. When fractionated according to particle density, C and N concentration (per g fraction) and C/N of these soil organo-mineral particles decrease with increasing particle density across soils of widely divergent texture, mineralogy, location, and management. The variation in particle density is explained potentially by two factors: (1) a decrease in the mass ratio of organic to mineral phase of these particles, and (2) variations in density of the mineral phase. The first explanation implies that the thickness of the organic accumulations decreases with increasing particle density. The decrease in C/N can be explained at least partially by especially stable sorption of nitrogenous N-containing compounds (amine, amide, and pyrrole) directly to mineral surfaces, a phenomenon well documented both empirically and theoretically. These peptidic compounds, along with ligand-exchanged carboxylic compounds, could then form a stable inner organic layer onto which other organics could sorb more readily than onto the unconditioned mineral surfaces (“onion” layering model).To explore mechanisms underlying this trend in C concentration and C/N with particle density, we sequentially density fractionated an Oregon andic soil at 1.65, 1.85, 2.00, 2.28, and 2.55 g cm⁻³ and analyzed the six fractions for measures of organic matter and mineral phase properties.All measures of OM composition showed either: (1) a monotonic change with density, or (2) a monotonic change across the lightest fractions, then little change over the heaviest fractions. Total C, N, and lignin phenol concentration all decreased monotonically with increasing density, and ¹⁴C mean residence time (MRT) increased with particle density from ca. 150 years to >980 years in the four organo-mineral fractions. In contrast, C/N, ¹³C and ¹⁵N concentration all showed the second pattern. All these data are consistent with a general pattern of an increase in extent of microbial processing with increasing organo-mineral particle density, and also with an “onion” layering model.X-ray diffraction before and after separation of magnetic materials showed that the sequential density fractionation (SDF) isolated pools of differing mineralogy, with layer-silicate clays dominating in two of the intermediate fractions and primary minerals in the heaviest two fractions. There was no indication that these differences in mineralogy controlled the differences in density of the organo-mineral particles in this soil. Thus, our data are consistent with the hypothesis that variation in particle density reflects variation in thickness of the organic accumulations and with an “onion” layering model for organic matter accumulation on mineral surfaces. However, the mineralogy differences among fractions made it difficult to test either the layer-thickness or “onion” layering models with this soil. Although SDF isolated pools of distinct mineralogy and organic-matter composition, more work will be needed to understand mechanisms relating the two factors.     

Arbuscular mycorrhizal fungi contribute to C and N enrichment of soil organic matter in forest soils

Increasing evidence suggests that accretion of microbial turnover products is an important driver for isotopic carbon (C) and nitrogen (N) enrichment of soil organic matter (SOM). However, the exact contribution of arbuscular mycorrhizal fungi (AMF) to soil isotopic patterns remains unknown. In this study, we compared ¹³C and ¹⁵N patterns of glomalin-related soil protein (GRSP), which includes a main fraction derived from AMF, litter, and bulk soil in four temperate rainforests. GRSP was an abundant C and N pool in these forest soils, showing significant ¹³C and ¹⁵N enrichment relative to litter and bulk soil. Hence, cumulative accumulation of recalcitrant AMF turnover products in the soil profile likely contributes to ¹³C and ¹⁵N enrichment in forest soils. Further research on the relationship between GRSP and AMF should clarify the exact extent of this process.     

Release of C and N from roots of peas and oats and their availability to soil microorganisms

Nutrient mobilisation in the rhizosphere is driven by soil microorganisms and controlled by the release of available C compounds from roots. It is not known how the quality of release influences this process in situ. Therefore, the present study was conducted to investigate the amount and turnover of rhizodeposition, in this study defined as root-derived C or N present in the soil after removal of roots and root fragments, released at different growth stages of peas (Pisum sativum L.) and oats (Avena sativa L.). Plants were grown in soil columns placed in a raised bed under outdoor conditions and simultaneously pulse labelled in situ with a ¹³C-glucose-¹⁵N-urea solution using a stem feeding method. After harvest, ¹³C and ¹⁵N was recovered in plant parts and soil pools, including the microbial biomass. Net rhizodeposition of C and N as a percentage of total plant C and N was higher in peas than in oats. Moreover, the C-to-N ratio of the rhizodeposits was lower in peas, and a higher proportion of the microbial biomass and inorganic N was derived from rhizodeposition. These results suggest a positive plant-soil feedback shaping nutrient mobilisation. This process is driven by the C and N supply of roots, which has a higher availability in peas than in oats.     

Natural 15N and 13C abundance in Andisols influenced by long-term fertilization management in Japan

Two field experiments were conducted on Andisols in Japan to evaluate the changes in the natural ¹⁵N and ¹³C abundance in the soil profile and to determine whether the values of δ¹⁵N could be used as an indicator of fertilizer sources or fertilizer fate. The 6-year experiment conducted at the National Agricultural Research Center (NARC) consisted of the following treatments: application of swine compost (COMPOST), slow-release nitrogen fertilizer (SRNF), readily available nitrogen fertilizer (RANF), and absence of fertilization (CONTROL). Experimental plots located at the Nippon Agricultural Research Institute (NARI) received cattle compost at different rates for 12 years; a forest soil at this site was sampled for comparison. Swine compost application led to a considerable change in the δ¹⁵N distribution pattern in the soil profile, with the highest δ¹⁵N values recorded in the top 20 cm layers of the COMPOST plot, decreasing in the sequence of CONTROL >- RANF > SRNF, mainly due to the relatively high δ¹⁵N value of swine compost and its subsequent decomposition. In contrast, SRNF application resulted in the lowest δ¹⁵N values in soil, indicating the presence of negligible nitrogen losses relative to input and low nitrogen cycling rates. Values of δ¹⁵N increased with compost application rates at NARI. In the leachate collected at 1-m depth, the δ¹⁵N values decreased in the sequence of COMPOST > RANF ≥ CONTROL > SRNF. The δ¹³C values in soil peaked in the 40–60 cm layers for all the fertilizers. The δ¹³C value was lowest in forest soil due to the presence of plant residues in soil organic matter. These results indicated that the δ¹⁵N values in the upper soil layers or leachate may enable to detect pollution sources of organic or inorganic nitrogen qualitatively in Andisols.     

Rhizosphere activity, grass species and N availability effects on the soil C and N cycles

We examined whether grass species and soil nitrogen (N) availability could enhance Carbon (C) and N turnover during root litter decay in grassland. Three species with increasing competitiveness (Festuca ovina, Dactylis glomerata and Lolium perenne) were grown at two N fertiliser levels in an undisturbed grassland soil, in which soil organic fractions derived for the last 9 years from Lolium root litter which was ¹³C-depleted. During the subsequent experimental year, the C turnover was calculated using the respective δ¹³C values of the old and new C in the root phytomass, in two Particulate Organic Matter (POM) fractions above 200 μm and in the lightest part of the aggregated soil fraction between 50 and 200 μm. Soil N availability was monitored during the regrowth periods with ion exchange resins (IER). The C decay rates of each particle size fraction were calculated with a simple mechanistic model of C dynamics. The N mineralisation immobilisation turnover (MIT) was characterised by dilution of ¹⁵N-labelled fertiliser in the N harvestThe C:N ratio and the residence time of C in the fractions decreased with particle size. The presence of a grass rhizosphere increased the decay rate of old C. Accumulation of new C in particle size fractions increased with species competitiveness and with N supply. Species competitiveness increased C turnover in the aggregated fraction, as a result of greater accumulation of new C and faster decay of old C. Fertiliser N increased N turnover and C mineralisation in the SOM. Species competitiveness decreased soil -N exchanged with the IER and increased dissolved organic C (DOC) content. The nature of the current rhizosphere is thus an important factor driving C and N transformations of the old root litter, in relation with grass species strategy. Plant competitiveness may stimulate the C and N turnover in the more evolved SOM fractions in a similar way to the mineral N supply.     

Plant nitrate use in deciduous woodland: the relationship between leaf N, N natural abundance of forbs and soil N mineralisation

Our aim was to study whether the in situ natural abundance ¹⁵N (δ¹⁵N)-values and N concentration of understory plants were correlated with the form and amount of mineral N available in the soil. Also to determine whether such differences were related to earlier demonstrations of differences in biomass increase in the same species exposed to nutrient solutions with both and or to alone. Several studies show that the δ¹⁵N of in soil solution generally is isotopically lighter than the δ¹⁵N of due to fractionation during nitrification. Hence, it is reasonable to assume that plant species benefiting from in ecosystems without significant leaching or denitrification have lower δ¹⁵N-values in their tissues than species growing equally well, or better, on We studied the δ¹⁵N of six understory species in oak woodlands in southern Sweden at 12 sites which varied fivefold in potential net N mineralisation rate The species decreased in benefit from in the following order: Geum urbanum, Aegopodium podagraria, Milium effusum, Convallaria majalis, Deschampsia flexuosa and Poa nemoralis. Four or five species demonstrated a negative correlation between and leaf δ¹⁵N and a positive correlation between and leaf N concentration. In wide contrast, only D. flexuosa, which grows on soils with little nitrification, showed a positive correlation between and the leaf N concentration and δ¹⁵N-value. Furthermore, δ¹⁵N of plants from the field and previously obtained indices of hydroponic growth on relative to were closely correlated at the species level. We conclude that δ¹⁵N may serve as a comparative index of uptake of among understory species, preferably in combination with other indices of N availability. The use of δ¹⁵N needs careful consideration of known restrictions of method, soils and plants.     

Uptake of carbon and nitrogen through roots of rice and corn plants,grown in soils treated with 13C and 15N dual-labeled cattle manure compost

Nitrogen and carbon dynamics in paddy and upland soils for rice cultivation and in upland soil for corn cultivation was investigated by using ¹³C and ¹⁵N dual-labeled cattle manure compost (CMC). In a soil with low fertility, paddy and upland rice took up carbon and nitrogen from the CMC at rates ranging from 0.685 to 1.051% of C and 17.6–34.6% of N applied. The ¹³C concentration was much higher in the roots than in the plant top, whereas the ¹⁵N concentration differed slightly between them, indicating that organic carbon taken up preferentially accumulated in roots. The ¹³C recovery in the plant top tended to be higher in upland soil than in paddy soil, whereas ¹⁵N applied was recovered at the same level in both paddy and upland soils. In the experiment with organic farming soil, paddy rice took up C and N from the CMC along with plant growth and the final recovery rates of ¹³C and ¹⁵N were 2.16 and 17.2% of C and N applied. In the corn experiment, a very large amount of carbon from the CMC was absorbed, accounting for at least 7 times value for rice. The final uptake rates of ¹³C and ¹⁵N reached about 13 and 10% of C and N applied, respectively. Carbon emission from the CMC sharply increased by 2 weeks after transplanting and the nitrogen emission was very low. It is concluded that rice and corn can take up an appreciable level of carbon and nitrogen from the CMC through roots.     

Investigation of the effect of ammonium sulfate on populations of ambient methane oxidising bacteria by C-labelling and GC/C/IRMS analysis of phospholipid fatty acids

The oxidation of atmospheric methane by methanotrophic bacteria residing in soils constitutes an important terrestrial methane sink with previous studies having revealed the inhibition of microbially mediated methane oxidation in the presence of salt ions. The bacteria responsible for ambient methane oxidation are not amenable to currently available methods of culturing, resulting in the need for a method of in situ analysis. A combination of phospholipid fatty acid (PLFA) analysis and stable isotopic labelling has been employed in this investigation as a means of cultivation-independent bacterial analysis. Soil samples were treated with an ammonium sulfate solution at a concentration that was known to inhibit methane oxidation or with distilled water, serving as a control, and incubated with ¹³C-labelled methane. PLFAs were analysed by GC/C/IRMS in order to determine their ¹³C content and, hence, the PLFA distribution of the methane oxidising bacteria. Ammonium sulfate treatment reduced the amount of ¹³C incorporated into the majority of PLFAs except the i17:0 PLFA in the presence of high concentrations of methane. These results implied a shift in the composition of the methane oxidising bacterial community in the soils treated with ammonium ions, with the treatment appearing to suppress one group of organisms more than another.     

C and N turnover in structurally intact soils of different texture

The turnover of native and applied C and N in undisturbed soil samples of different texture but similar mineralogical composition, origin and cropping history was evaluated at −10 kPa water potential. Cores of structurally intact soil with 108, 224 and 337 g clay kg⁻¹ were horizontially sliced and ¹⁵N-labelled sheep faeces was placed between the two halves of the intact core. The cores together with unamended treatments were incubated in the dark at 20 °C and the evolution of CO₂-C determined continuously for 177 d. Inorganic and microbial biomass N and ¹⁵N were determined periodically. Net nitrification was less in soil amended with faeces compared with unamended soil. When adjusted for the NO₃-N present in soil before faeces was applied, net nitrification became negative indicating that NO₃-N had been immobilized or denitrified. The soil most rich in clay nitrified least N and ¹⁵N. The amounts of N retained in the microbial biomass in unamended soils increased with clay content. A maximum of 13% of the faeces ¹⁵N was recovered in the microbial biomass in the amended soils. CO₂-C evolution increased with clay content in amended and unamended soils. CO₂-C evolution from the most sandy soil was reduced due to a low content of potentially mineralizable native soil C whereas the rate constant of C mineralization rate peaked in this soil. When the pool of potentially mineralizable native soil C was assumed proportional to volumetric water content, the three soils contained similar proportions of potentially mineralizable native soil C but the rate constant of C mineralization remained highest in the soil with least clay. Thus although a similar availability of water in the three soils was ensured by their identical matric potential, the actual volume of water seemed to determine the proportion of total C that was potentially mineralizable. The proportion of mineralizable C in the faeces was similar in the three soils (70% of total C), again with a higher rate constant of C mineralization in the soil with least clay. It is hypothesized that the pool of potentially mineralizable C and C rate constants fluctuate with the soil water content.     

Depth dynamics of soil N contents and natural abundances of 15N after 43 years of long-term fertilization and liming in sub-tropical Alfisol

The aim of this study was to understand impacts of long-term (43 years) fertilization on soil aggregation, N accumulation rates and δ¹⁵N in surface and deep layers in an Alfisol. Soil samples from seven treatments were analysed for N stocks, aggregate-associated N in 0–30 cm and the changes in δ¹⁵N in 0–90 cm depths. The treatments were: unfertilized control (control); recommended N dose (N); recommended N and phosphorus doses (NP); recommended N, P and potassium doses (NPK); 150% of recommended N, P and K doses (150% NPK); NPK + 10 Mg FYM ha^?1 (NPK + FYM) and NPK + 0.4 Mg lime ha^?1 (NPK + L). Results revealed that plots under NPK + FYM had ~39% higher total N concentrations than NPK + L in 0–30 cm soil layers. In NPK + L, macro-aggregates had 35 and 11% and microaggregates had 20 and 9% lower δ¹⁵N values than NPK + FYM in 0–15 and 15–30 cm soil layers, respectively. However, plots receiving NPK + FYM had ~39% greater deep soil (30–90 cm) N accumulation than NPK + L. These results would help understanding N supplying capacity by long-term fertilization and assist devising N management strategies in sub-tropical acidic Alfisols.     

Tracing the source and fate of dissolved organic matter in soil after incorporation of a C labelled residue: A batch incubation study

While dissolved organic matter (DOM) in soil solution is a small but reactive fraction of soil organic matter, its source and dynamics are unclear. A laboratory incubation experiment was set up with an agricultural topsoil amended with ¹³C labelled maize straw. The dissolved organic carbon (DOC) concentration in soil solution increased sharply from 25 to 186 mg C L⁻¹ 4 h after maize amendment, but rapidly decreased to 42 mg C L⁻¹ and reached control values at and beyond 2 months. About 65% of DOM was straw derived after 4 h, decreasing to 29% after one day and only 1.3% after 240 days. A significant priming effect of the straw on the release of autochthonous DOM was found. The DOM fractionation with DAX-8 resin revealed that 98% of the straw derived DOM was hydrophilic in the initial pulse while this hydrophilic fraction was 20-30% in control samples. This was in line with the specific UV absorbance of the DOM which was significantly lower in the samples amended with maize residues than in the control samples. The δ¹³C of the respired CO₂ matched that of DOC in the first day after amendment but exceeded it in following days. The straw derived C fractions in respired CO₂ and in microbial biomass were similar between 57 and 240 days after amendment but were 3-10 fold above those in the DOM. This suggests that the solubilisation of C from the straw is in steady state with the DOM degradation or that part of the straw is directly mineralised without going into solution. This study shows that residue application releases a pulse of hydrophilic DOM that temporarily (<3 days) dominates the soil DOM pool and the degradable C. However, beyond that pulse the majority of DOM is derived from soil organic matter and its isotope signature differs from microbial biomass and respired C, casting doubt that the DOM pool in the soil solution is the major bioaccessible C pool in soil.