Endogeic earthworms alter carbon translocation by fungi at the soil-litter interface

The effect of endogeic earthworms (Octolasion tyrtaeum (Savigny)) on the translocation of litter-derived carbon into the upper layer of a mineral soil by fungi was investigated in a microcosm experiment. Arable soil with and without O. tyrtaeum was incubated with ¹³C/¹⁵N-labelled rye leaves placed on plastic rings with gaze (64 μm mesh size) to avoid incorporation of leaves by earthworms. The plastic rings were positioned either on or 3 cm above the soil surface, to distinguish between biotic and chemical/physical translocation of nutrients by fungi and leaching.Contact of leaves to the soil increased ¹³C translocation, whereas presence of O. tyrtaeum reduced the incorporation of ¹³C into the mineral soil in all treatments. Although biomass of O. tyrtaeum decreased during the experiment, more ¹³C and ¹⁵N was incorporated into earthworm tissue in treatments with contact of leaves to the soil. Contact of leaves to the soil and the presence of O. tyrtaeum increased cumulative ¹³CO₂-C production by 18.2% and 14.1%, respectively.The concentration of the fungal bio-indicator ergosterol in the soil tended to be increased and that of the fungal-specific phospholipid fatty acid 18:2ω6 was significantly increased in treatments with contact of leaves to the soil. Earthworms reduced the concentration of ergosterol and 18:2ω6 in the soil by 14.0% and 43.2%, respectively. Total bacterial PLFAs in soil were also reduced in presence of O. tyrtaeum, but did not respond to the addition of the rye leaves. In addition, the bacterial community in treatments with O. tyrtaeum differed from that without earthworms and shifted towards an increased dominance of Gram-negative bacteria.The results indicate that litter-decomposing fungi translocate litter-derived carbon via their mycelial network in to the upper mineral soil. Endogeic earthworms decrease fungal biomass by grazing and disruption of fungal hyphae thereby counteracting the fungal-mediated translocation of carbon in soils.     

Markers of Soil Organic Matter Transformation in Permafrost Peat Mounds of Northeastern Europe

For the paleoreconstruction of permafrost peat mounds and the identification of plant communities participating in the formation of peat, the contents of n-alkanes (C₂₀–C₃₃) have been determined, and relative changes in the stable isotope compositions of carbon and nitrogen and the C/N ratio have been analysed. Several indices (CPI_alkanes, P_aq, P_wax) have been calculated to assess the degree of decomposition of the peats studied and the contributions of different plant species to their formation. It has been found that shortand long-chain n-alkanes are concentrated in high-moor peat, while medium-chain alkanes are typical for transitional peat. Integrated analysis of the studied markers has shown that the botanical and material composition of peat, anaerobic conditions of bog formation, and permafrost play an important role in the preservation of organic carbon in permafrost peat mounds. Alternation of plant associations is the main reason for changes in n-alkane concentrations, C/N ratios, and δ13C values.     

Health of remnant woodlands in fragments under distinct grazing regimes

Fragmented remnant woodlands in agricultural landscapes are of high conservation value world-wide. Many eucalypts in agricultural landscapes of Australia are in decline. We aimed to investigate nutrient enrichment as a process that may contribute to eucalypt decline. We studied remnant woodlands that had been exposed to distinct recent and current livestock grazing treatments: Currently Intense Grazed; Recently Intense Grazed (until 3 years ago); Recent Intermediate Grazed; and Recent Lightly Grazed by livestock. We assessed soil nutrient status and penetrability, eucalypt foliar nutrition and stable isotope ratios for N and C, attributes of understorey vegetation, and tree health. Soils of the Currently Intense Grazed treatment had high levels of ammonium and Colwell-P. Total N, P, C:N ratio and soil penetrability were generally high in Currently Intense Grazed and Recently Intense Grazed treatments relative to Recent Intermediate Grazed and Recent Lightly Grazed treatments. Foliar N, N stable isotope ratios, P and carbon stable isotope ratios (δ¹³C) were generally higher (less negative δ¹³C) in trees on Currently Intense Grazed and Recently Intense Grazed treatments than in trees on Recent Intermediate Grazed and Recent Lightly Grazed treatments. Soil surface litter, tall and low shrubs and rock were positively correlated with tree health. Grasses and eucalypt foliar N, P and δ¹³C were negatively correlated with tree health. Soil nutrient enrichment increased with increasing grazing intensity and was associated with increased weed invasion and with poor tree health that was in turn correlated to increased foliar N and P and less negative δ¹³C in woodland trees in this study. We argue that minimising soil nutrient enrichment of fragmented remnant woodlands is important, given the association of elevated soil nutrition with poor tree health, to ensure the persistence of eucalypts in agricultural landscapes.     

Short-term C4 plant Spartina alterniflora invasions change the soil carbon in C3 plant-dominated tidal wetlands on a growing estuarine Island

Spartina alterniflora is an invasive C₄ perennial grass, native to North America, and has spread rapidly along the east coast of China since its introduction in 1979. Since its intentional introduction to the Jiuduansha Island in the Yangtze River estuary, Spartina alterniflora community has become one of the dominant vegetation types. We investigated the soil carbon in the Spartina alterniflora community and compared it with that of the native C₃Scirpus mariqueter community by measuring total soil carbon (TC), soil organic carbon (SOC), total soil nitrogen (TN), and the stable carbon isotope composition (δ¹³C) of various fractions. TC and SOC were significantly higher in Spartina alterniflora in the top 60 cm of soil. However, there was no significant difference in soil inorganic carbon (IC) between the two communities. Stable carbon isotopic analysis suggests that the fraction of SOC pool contributed by Spartina alterniflora varied from 0.90% to 10.64% at a soil depth of 0-100 cm with a greater percentage between 20 and 40 cm deep soils. The δ¹³C decreased with increasing soil depth in both communities, but the difference in δ¹³C among layers of the top 60 cm soil was significant (p<0.05), while that for the deeper soil layers (>60 cm) was not detected statistically. The changes in δ¹³C with depth appeared to be associated with the small contribution of residues from Spartina alterniflora at greater soil depth that was directly related to the vertical root distribution of the species.     

Variability in pattern and hydrogen isotope composition (δ2H) of long-chain n-alkanes of surface soils and its relations to climate and vegetation characteristics: A meta-analysis

The average chain length (ACL), carbon preference index (CPI), and hydrogen isotope composition (δ²H) of long-chain n-alkanes in sediments have been used to retrieve information about the paleoclimate. Despite their importance as in-between media from leaves to sediments, n-alkanes of surface soils have not been systematically analyzed at large scale. Such an investigation of the spatial variation of n-alkane properties in soil and their dependence on climatic and botanic (e.g., vegetation type) factors could provide a rationale for a better estimation of the past environment. We synthesized the patterns and δ²H of long-chain n-alkanes in soil (δ²H^n-alkanes) with regard to vegetation types (cropland, grassland, shrubland, and woodland) and environmental factors using data from peer-reviewed papers. Our results showed that the ACL and CPI of soil C₂₇–C₃₃ n-alkanes were not suitable indicators for differentiating vegetation types at large scale; instead, ACL significantly correlated with water conditions such as mean annual precipitation (MAP) and Palmer drought severity index (PDSI), and CPI significantly correlated with temperature without significant influence of vegetation type. The variation (i.e., standard deviation) of fractionation between the δ²H values in annual precipitation and in soil n-alkanes (ε_rain-soil) was smaller than that reported in leaves; therefore, soils were better suited to quantifying the general growing conditions of plants at a certain site. The fractionation ε_rain-soil correlated with climatic conditions as described by the PDSI and relative humidity (RH). This correlation agreed with the change in leaf water enrichment with changing RH taken from the literature and was independent of the vegetation type at large scale. This meta-analysis may provide useful information for the variations of the patterns and δ²H_n-alkanes values in surface soils.     

Plant species influence microbial diversity and carbon allocation in the rhizosphere

Plant species effects on microbial communities are attributed to changes in microbial community composition and biomass, and may depend on plant species specific differences in the quality of resources (carbon) inputs. We examined the idea that plant-soil feedbacks can be explained by a chance effect, which is the probability of a highly productive or keystone plant species is present in the community and will influence the functions more than the number of species per se. A ¹³C pulse labelling technique was applied to three plant species and a species mixture in a greenhouse experiment to examine the carbon flow from plants to soil microbial communities. The ¹³C label was given as CO₂ to shoots of a legume (Lotus corniculatus), a forb (Plantago lanceolata), a grass (Holcus lanatus) and a mixture of the three species. Microbial phospholipid fatty acids (PLFA) was analysed in order to determine the biomass and composition of the soil microbial community. The incorporation of the stable isotope into soil microorganisms was determined through GC-IRMS analyses of the microbial PLFAs. Plant species identity did not influence the microbial biomass when determined as total carbon of microbial phospholipid fatty acids. However, the labelled carbon showed that the grass monoculture (H. lanatus) and the plant mixture allocated more ¹³C into bacteria and actinomycete biomass than the other plant species. H. lanatus monocultures had also the highest amounts of ¹³C allocated to AM-fungi and saprophytic fungi. The carbon allocation from plants to soil microorganisms in a plant species mixture can thus be explained by the presence of a highly productive species that influence soil functions.     

Five years of simulated atmospheric nitrogen deposition have only subtle effects on the fate of newly synthesized carbon in Calluna vulgaris and Eriophorum vaginatum

To understand the implications of atmospheric nitrogen deposition on carbon turnover in peatlands, we conducted a ¹³C pulse labeling experiment on Calluna vulgaris and Eriophorum vaginatum already receiving long-term (5 years) amendments of 56 kg N ha⁻¹ y⁻¹ as ammonium or nitrate. We examined shoot tissue retention, net ecosystem respiration returns of the ¹³C pulse, and soil porewater DOC content under the two species. ¹³C fixation in Eriophorum leaves was enhanced with nitrogen addition and doubled with nitrate supply. This newly fixed C appeared to be relocated below-ground faster with nitrogen fertilization as respiration returns were unaffected by N inputs. By contrast, increases in ¹³C fixation were not observed in Calluna. Instead, net ecosystem respiration rates over Calluna increased with N fertilization. There was no significant label incorporation into DOC, suggesting a conservative strategy of peatland vegetation regarding allocation of C through root exudation. Greater concentrations of total DOC were identified with nitrate addition in Calluna. Given the long-term nature of the experiment and the high N inputs, the overall impacts of nitrogen amendments on the fate of recently synthesized C in Eriophorum and Calluna in this ombrotrophic peatland were surprisingly more moderate than originally hypothesized. This may be due to N being effectively retained within the bryophyte layer, thus limiting, and delaying the onset of, below-ground effects.     

Bacterial populations assimilating carbon from C-labeled plant residue in soil: Analysis by a DNA-SIP approach

In this study, ¹³C-labeled rice callus was prepared as a model material for rice straw and was subjected to a DNA-SIP (stable isotope probing) experiment in which the bacterial population was monitored in a soil sample containing decomposing dried callus. Rice callus (¹³C = 78%) contained the more water-soluble organic carbon and less cellulose and lignin carbon than rice straw. The callus in the soil was 37% decomposed after 56 d of incubation in upland moisture conditions. PCR-DGGE analysis demonstrated that the bacterial community in the soil with the callus changed over time, showing a distinct difference between the first (up to 7 d) and second (14 d and later) stages. After isopycnic centrifugation, DNA in the fractions with a buoyant density between 1.759 and 1.734 g ml⁻¹ was subjected to population analysis (¹³C-assimilating populations). Diverse groups of bacterial sequences were retrieved from the ¹³C-labeled DNA fractions: Actinobacteria, Bacilli, γ-Proteobacteria, Chloroflexi, Sphingobacteria, Flavobacteria, Clostridia, Acidobacteria, Cyanobacteria and Candidate Division. Bacilli were detected mainly in the first stage, and Actinobacteria were detected throughout the incubation period. Several DGGE bands in the light fractions became more prominent in the soil with callus, which suggested that the addition of callus promoted the growth of bacteria that fed on soil organic matter, including α-Proteobacteria, γ-Proteobacteria, Bacilli, Actinobacteria, Nitrospira and Gemmatimonadetes.     

Evaluating N/N and C/C isotope ratio analysis to investigate trophic relationships of elaterid larvae (Coleoptera: Elateridae)

Carbon and nitrogen isotope ratios in consumer tissues can be used to analyse the diet and trophic level of soil animals. However, life history traits may significantly influence stable isotope patterns. We evaluated in a series of experiments how stable isotope ratios of carbon (¹³C/¹²C) and nitrogen (¹⁵N/¹⁴N) at natural abundance can be used to study the diet and trophic position of long-lived macro-invertebrates, elaterid larvae, which are major below-ground herbivores. Small, but significant differences in δ¹³C signatures were found between the larvaes’ anterior and posterior body segments, whereas exuvia reflected the body's overall isotopic composition. The species-specific trophic shift (±SE) in δ¹⁵N for Agriotes obscurus and Agriotes sputator (1.62±0.24‰ and 1.08±0.27‰, respectively) was significantly lower than “mean enrichment estimates” reported in the literature, showing the limited applicability of such generalised estimates in studies of invertebrate trophic ecology. To avoid false-positive assignments to two trophic levels due to variation in δ¹⁵N values, a minimum sample size of three and five individuals for A. obscurus and A. sputator, respectively, was needed to reduce this risk to below α=5%. Keeping elaterid larvae for up to 128 days without food did not affect their isotopic signatures, in contrast to previous studies on starving animals. Switching wireworms to isotopically different diets induced changes in their isotopic signatures within 2 weeks. Changes, however, were significant only when the isotopic difference between diets was large. We conclude that experimental studies evaluating how specific life history traits affect stable isotope signatures in consumers have to precede any interpretation of stable isotope data gathered in the field.     

A GC/MS method for the assessment of N and C incorporation into soil amino acid enantiomers

Identifying the transformation process of amino acid enantiomers was essential to probe into the fate, turnover and aging of soil nitrogen due to their important roles in the biogeochemical cycling. If this can be achieved by differentiating between the newly biosynthesized and the inherent compounds in soil, then the isotope tracer method can be considered most valid. We thereby developed a gas chromatography/mass spectrometry (GC/MS) method to trace the ¹⁵N or ¹³C isotope incorporation into soil amino acid enantiomers after being incubated with ¹⁵NH₄⁺ or U-¹³C-glucose substrates. The most significant fragments (F) as well as the related minor ions were monitored by the full scan mode and the isotope enrichment in amino acids was estimated by calculating the atom percentage excess (APE). ¹⁵NH₄⁺ incorporation was evaluated according to the relative abundance increase of m/z F+1 to F for neutral and acidic amino acids and F+2 to F (mass 439) for lysine. The assessment of ¹³C enrichment in soil amino acids was more complicated than that of ¹⁵N due to multi-carbon atoms in amino acid molecules. The abundance ratio increment of m/z F+n to F (n is the original skeleton carbon number in each fragment) indicated the direct conversion from the added glucose to amino acids, but the total isotope incorporation from the added ¹³C can only be calculated according to all target isotope fragments, i.e. the abundance ratio increment summation from m/z (Fa+1) through m/z (Fa+T) represented the total incorporation of the added ¹³C (Fa is the fragment containing all original skeleton carbons and T is the carbon number in the amino acid molecule). This method has a great advantage especially for the evaluation of high-abundance isotope enrichment in organic compounds compared with GC/C/IRMS. And in principle, this technique is also valid for amino acids besides enantiomers if stereoisomers are not concerned. Our assessment approach could shine a light on investigating the biochemical mechanism of microbial transformation of N and C in soils of terrestrial ecosystem.     

Examining the Microbial Degradation of Naphthenic Acids Using Stable Isotope Analysis of Carbon and Nitrogen

Naphthenic acids (NAs) are a complex group of naturally occurring oil sands constituents that constitute a significant portion of the dissolved organic carbon (DOC) pool available for microbial degradation in the process-related waste water associated with oil sands mine sites. One approach to understanding the biological fate of oil sands process-derived carbon and nitrogen in aquatic reclamation of the mine sites involves the use of stable isotope analyses. However, for stable isotope analyses to be useful in such field-based assessments, there is a need to determine how microbial degradation of a complex mixture of NAs might change the stable isotope values (δ ¹³C, δ ¹⁵N). In batch cultures and semi-continuous laboratory microcosms, utilization of a commercial mixture of NAs by oil sands-derived microbial cultures resulted in microbial biomass that was similar or slightly ¹³C enriched (1.4‰ to 3.0‰) relative to the DOC source, depending on the length of incubation. Utilization of a NA-containing extract of oil sands processed water resulted in greater ¹³C enrichment of microbial biomass (8.5‰) relative to the DOC source. Overall, the δ ¹³C of the DOC comprised of complex mixtures of NAs showed minimal change (?0.5‰ to ?0.1‰) during the incubation period whereas the δ ¹³C of the dissolved inorganic carbon (DIC) was more variable (?5.0‰ to +5.4‰). In tests where the concentration of available nitrogen was increased, the final biomass values were ¹⁵N enriched (3.8‰ to 8.4‰) relative to the initial biomass. The isotope trends established in this study should enhance our ability to interpret field-based data from sites with hydrocarbon contamination, particularly in terms of carbon source utilization and ¹⁵N enrichment.     

Sensitivity analysis and quantification of uncertainty for isotopic mixing relationships in carbon cycle research

Quantifying and understanding the uncertainty in isotopic mixing relationships is critical to isotopic applications in carbon cycle studies at all spatial and temporal scales. Studies that depend on stable isotope approaches must also address quantification of uncertainty for parameters derived from isotopic studies. An important application of isotopic mixing relationships is determination of the isotopic content of ecosystem respiration (δ¹³C_S) via an inverse relationship (a Keeling plot) between atmospheric CO₂ concentrations ([CO₂]) and carbon isotope ratios of CO₂ (δ¹³C). Alternatively, a linear relationship between [CO₂] and the product of [CO₂] and δ¹³C (a Miller/Tans plot) can also be applied.We used three datasets of [CO₂] and δ¹³C in air to examine contrasting approaches to determine δ¹³C_S and its uncertainty. These datasets were from the Niwot Ridge, Colorado, AmeriFlux site, the Biosphere-Atmosphere Stable Isotope Network (BASIN), and from the Grünschwaige Grassland Research Station in Germany. The analysis of this data included Keeling plots and Miller/Tans plots fit with both Model I (ordinary least squares) and Model II regressions (geometric mean regression and orthogonal distance regression).Our analysis confirms previous observations that increasing the range of the measurements ([CO₂] range) used for a mixing line reduces the uncertainty associated with δ¹³C_S. Using a Model II regression technique to determine δ¹³C_S introduces a negatively skewed bias in δ¹³C_S which is especially significant for small [CO₂] ranges. This bias arises from comparatively greater variability in the dependent variable than the independent variable for a linear regression. For carbon isotope studies, uncertainty in the isotopic measurements has a greater effect on the uncertainty of δ¹³C_S than the uncertainty in [CO₂]. As a result, studies that estimate parameters via a Model II regression technique maybe biased in their conclusions. In contrast to earlier studies, we advocate Model I (ordinary least squares) regression to calculate δ¹³C_S and its uncertainty. Reducing the uncertainty of isotopic measurements reduces the uncertainty of δ¹³C_S, even when the [CO₂] range of samples is small (<20 ppm). As a result, improvement in isotope (rather than [CO₂]) measuring capability is presently needed to substantially reduce uncertainty in δ¹³C_S. We find for carbon isotope studies no inherent advantage or disadvantage to using either a Keeling or Miller/Tans approach to determine δ¹³C_S. We anticipate that the mathematical methods developed in this paper can be applied to other applications where linear regression is utilized.     

Relationships between C and N availability, substrate age, and natural abundance C and N signatures of soil microbial biomass in a semiarid climate

Soil microbial organisms are central to carbon (C) and nitrogen (N) transformations in soils, yet not much is known about the stable isotope composition of these essential regulators of element cycles. We investigated the relationship between C and N availability and stable C and N isotope composition of soil microbial biomass across a three million year old semiarid substrate age gradient in northern Arizona. The δ¹⁵N of soil microbial biomass was on average 7.2‰ higher than that of soil total N for all substrate ages and 1.6‰ higher than that of extractable N, but not significantly different for the youngest and oldest sites. Microbial ¹⁵N enrichment relative to soil extractable and total N was low at the youngest site, increased to a maximum after 55,000 years, and then decreased slightly with age. The degree of ¹⁵N enrichment of microbial biomass correlated negatively with the C:N mass ratio of the soil extractable pool. The δ¹³C signature of soil microbial biomass was 1.4‰ and 4.6‰ enriched relative to that of soil total and extractable pools respectively and showed significant differences between sites. However, microbial ¹³C enrichment was unrelated to measures of C and N availability. Our results confirm that ¹⁵N, but not ¹³C enrichment of soil microbial biomass reflects changes in C and N availability and N processing during long-term ecosystem development.     

Fate of gram-negative bacterial biomass in soil—mineralization and contribution to SOM

Soil microorganisms contribute to the formation of non-living soil organic matter (SOM) by metabolic transformation of plant-derived material. After cell death, their biomass components with a specific molecular character become incorporated into SOM imprinting its chemical properties, although this process has not yet been quantified. In order to elucidate the contribution to SOM formation, we investigated the fate of gram-negative bacterial model biomass (Escherichia coli usually introduced into soil with manure or feces) during incubation of soil with isotopically (¹³C) and genetically (lux gene) labeled cells. The decline of living cells was monitored by the loss of bioluminescence. The carbon turnover and mineralization was balanced by bulk soil stable isotope analysis, and the persistence of nucleic acids was investigated by PCR amplification of the lux gene. During incubation, the number of viable E. coli cells decreased rapidly (99.9% within the first 42 d) serving as substrate for other microorganisms or for the formation of SOM, and bioluminescent cells could only be detected during the first 56 d. However, the lux gene was still detected after 224 d, which indicates stabilization of DNA in SOM. Although the survival of E. coli in soil is limited, only about 65% of the added labeled biomass carbon was mineralized to ¹³CO₂ and 51% remained in soil after 224 d with an average ¹³C recovery of 117%. The amount of ¹³C found in the PLFA representative of living cells had decreased to 25% of the initial value, suggesting a proportional decrease of the ¹³C in the soil microbial biomass. The extent of this decrease is higher than the mineralization of the bulk E. coli C and thus the difference of around 25% has to be stabilized as metabolites, or in non-living SOM. The data provide evidence that the genetic information and a considerable part of the carbon from dying bacterial biomass were retained in both the soil microbial food web and in non-living SOM.     

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.     

Roots from beech (Fagus sylvatica L.) and ash (Fraxinus excelsior L.) differentially affect soil microorganisms and carbon dynamics

Knowledge about the influence of living roots on decomposition processes in soil is scarce but is needed to understand carbon dynamics in soil. We investigated the effect of dominant deciduous tree species of the Central European forest vegetation, European beech (Fagus sylvatica L.) and European ash (Fraxinus excelsior L.), on soil biota and carbon dynamics differentiating between root- and leaf litter-mediated effects. The influence of beech and ash seedlings on carbon and nitrogen flow was investigated using leaf litter enriched in ¹³C and ¹⁵N in double split-root rhizotrons planted with beech and ash seedlings as well as a mixture of both tree species and a control without plants. Stable isotope and compound-specific fatty acid analysis (¹³C-PLFA) were used to follow the incorporation of stable isotopes into microorganisms, soil animals and plants. Further, the bacterial community composition was analyzed using pyrosequencing of 16S rRNA gene amplicons. Although beech root biomass was significantly lower than that of ash only beech significantly decreased soil carbon and nitrogen concentrations after 475 days of incubation. In addition, beech significantly decreased microbial carbon use efficiency as indicated by higher specific respiration. Low soil pH probably increased specific respiration of bacteria suggesting that rhizodeposits of beech roots induced increased microbial respiration and therefore carbon loss from soil. Compared to beech δ¹³C and δ¹⁵N signatures of gamasid mites in ash rhizotrons were significantly higher indicating higher amounts of litter-derived carbon and nitrogen to reach higher trophic levels. Similar δ¹³C signatures of bacteria and fine roots indicate that mainly bacteria incorporated root-derived carbon in beech rhizotrons. The results suggest that beech and ash differentially impact soil processes with beech more strongly affecting the belowground system via root exudates and associated changes in rhizosphere microorganisms and carbon dynamics than ash.     

Stable isotope labelling of earthworms can help deciphering belowground–aboveground interactions involving earthworms,mycorrhizal fungi,plants and aphids

Functional relationships between belowground detritivores and/or symbionts and aboveground primary producers and their herbivores are not well studied. In a factorial greenhouse experiment we studied interactions between earthworms (addition/no addition of Lumbricus terrestris; Clitellata: Lumbricidae) and arbuscular-mycorrhizal fungi (AMF; with/without inoculation of Glomus mosseae; Glomerales: Glomeraceae) on the leguminous herb Trifolium repens (Fabales: Fabaceae) and associated plant aphids (Aphis gossypii, A. craccivora; Hemiptera: Aphidoidea). In order to be able to trace organismic interactions, earthworms were dual-labelled with stable isotopes (¹⁵N-ammonium nitrate and ¹³C-glucose). We specifically wanted to investigate whether (i) isotopic signals can be traced from the labelled earthworms via surface castings, plant roots and leaves to plant aphids and (ii) these compartments differ in their incorporation of stable isotopes. Our results show that the tested organismic compartments differed significantly in their ¹⁵N isotope enrichments measured seven days after the introduction of earthworms. ¹⁵N isotope incorporation was highest in casts followed by earthworm tissue, roots and leaves, with lowest ¹⁵N signature in aphids. The ¹³C signal in roots, leaves and aphids was similar across all treatments and is for this reason not recommendable for tracing short-term interactions over multitrophic levels. AMF symbiosis affected stable isotope incorporation differently in different subsystems: the ¹⁵N isotope signature was higher below ground (in roots) but lower above ground (leaves and aphids) in AMF-inoculated mesocosms compared to AMF-free mesocosms (significant subsystem × AMF interaction). Aphid infestation was unaffected by AMF and/or earthworms. Generally, these results demonstrate that plants utilize nutrients excreted by earthworms and incorporate these nutrients into their roots, leaf tissue and phloem sap from where aphids suck. Hence, these results show that earthworms and plant aphids are functionally interlinked. Further, ¹⁵N-labelling earthworms may represent a promising tool to investigate nutrient uptake by plants and consequences for belowground-aboveground multitrophic interactions.     

Carbohydrate use and assimilation by litter and soil fungi assessed by carbon isotopes and BIOLOG assays

Soil fungi are integral to decomposition in forests and other habitats, yet identifying probable functional roles of different taxa is problematic. Here, we compared carbohydrate assimilation patterns derived from stable isotope analyses on cultures with patterns of metabolic activity measured on Biolog^® SF-P plates for 12 taxa of soil- and litter-inhabiting saprotrophic fungi isolated from Douglas-fir (Pseudotsuga menziesii) ecosystems. To determine the relative assimilation of carbon from malt extract versus sucrose by ¹³C stable isotope analyses, we cultured fungi with malt extract (consisting primarily of glucose and maltose) plus either C₃- or C₄-derived sucrose as carbon sources. Rhodotorula graminis and F. oxysporum assimilated the highest proportion of sucrose, a Mortierella isolate and an unidentified sterile isolate (FPC 341) assimilated the lowest proportion of sucrose, and remaining cultures assimilated similar and intermediate proportions of sucrose. On Biolog plates, low metabolic activity of Mortierella and FPC 341 on sucrose and R. graminis and F. oxysporum on maltose were qualitatively consistent with isotopic results. Assimilation of sucrose calculated isotopically was correlated with the ratio of sucrose: maltose activity calculated from Biolog assays (r²=0.45, P=0.0145, n=12). Metabolic activity on Biolog plates for six other common soil carbohydrates were also determined: glucose, fructose, galactose, cellobiose, lactose, and glycogen. Metabolic activity was greatest overall on maltose and glucose and lowest on fructose. Two of the isolates (Aspergillus flavus and F. oxysporum) had higher metabolic activity on the glucose-containing disaccharide cellobiose than on glucose, strongly suggesting preferential uptake of cellobiose compared to glucose and suggesting the potential ability to use cellulose. The high metabolic activity of these cultures on galactose, a primary constituent of hemicellulose, also suggested cellulolytic capabilities. Our results indicated that stable isotope studies and Biolog assays may provide complementary information to characterize metabolic potential of fungi in forest litter and soil.     

Carbon in tundra soils in the Lake Labaz region of arctic Siberia

Large amounts of carbon are stored in permafrost‐affected soils of the Arctic tundra. The quantity, distribution and composition of this carbon are important, because much of the carbon is likely to be released as a result of global warming. We have studied soils of the central Siberian Arctic to determine the carbon content and the nature of the organic matter by density fractionation, and ¹³C‐NMR‐ and ¹³C‐stable‐isotope analyses. There are pronounced differences in the profile and variations from place to place in the quantity and nature of soil organic matter. We estimated that the mean stock of carbon was 14.5 kg m^–2 within the active layer. We found a total of about 30.7 kg C m^–3 in the entire upper metre of the soils. Carbon of the tussock tundra showed strong vertical differentiation, with a large proportion comprising decomposed, recalcitrant compounds. We identified within the soil several zones of aerobe and anaerobe decomposition. Mobile carbon fractions have precipitated under the influence of low temperatures.     

Below ground carbon turnover and greenhouse gas exchanges in a sub-arctic wetland

Here we present results from a field experiment in a sub-arctic wetland near Abisko, northern Sweden, where the permafrost is currently disintegrating with significant vegetation changes as a result. During one growing season we investigated the fluxes of CO₂ and CH₄ and how they were affected by ecosystem properties, i.e., composition of species that are currently expanding in the area (Carex rotundata, Eriophorum vaginatum and Eriophorum angustifolium), dissolved CH₄ in the pore water, substrate availability for methane producing bacteria, water table depth, active layer, temperature, etc. We found that the measured gas fluxes over the season ranged between: CH₄ 0.2 and 36.1 mg CH₄ m⁻² h⁻¹, Net Ecosystem Exchange (NEE) −1000 and 1250 mg CO₂ m⁻² h⁻¹ (negative values meaning a sink of atmospheric CO₂) and dark respiration 110 and 1700 mg CO₂ m⁻² h⁻¹. We found that NEE, photosynthetic rate and CH₄ emission were affected by the species composition. Multiple stepwise regressions indicated that the primary explanatory variables for NEE was photosynthetic rate and for respiration and photosynthesis biomass of green leaves. The primary explanatory variables for CH₄ emissions were depth of the water table, concentration of organic acid carbon and biomass of green leaves. The negative correlations between pore water concentration and emission of CH₄ and the concentrations of organic acid, amino acid and carbohydrate carbon indicated that these compounds or their fermentation by-products were substrates for CH₄ formation. Furthermore, calculation of the radiative forcing of the species expanding in the area as a direct result of permafrost degradation and a change in hydrology indicate that the studied mire may act as an increasing source of radiative forcing in future.