Short-term dynamics of slurry-derived plant and microbial sugars in a temperate grassland soil as assessed by compound-specific delta13C analyses

In view of recent discussions about climate change and the anthropogenically enhanced greenhouse effect, the aim of this study was to determine the short-term carbon (C) dynamics in a grassland soil after slurry application. It is known that, depending on cultivation practices, agro-ecosystems can act either as sources or as sinks for atmospheric CO2. C3 and C4 slurries were applied, differing in their stable C isotope signature, to be able to differentiate between native (soil-inherent) and fresh (slurry-applied) C. Samples were taken from 0-2, 2-7.5 and 7.5-15 cm soil depths from 90 days before until 4 weeks after slurry application at various intervals. We carried out compound-specific stable isotope analysis (CSIA) of plant- (arabinose and xylose) and microbial-derived sugars (fucose and rhamnose). Up to 45% of the applied slurry-derived xylose was found in the 0-2 cm soil depth within 24 h after slurry application, with this figure decreasing rapidly and then increasing again towards the end of the experiment. Therefore, during the first phase of slurry incorporation, preferentially the soluble part of slurry entered the first 2 cm of soil while, after about 2 weeks, particulate slurry-derived organic matter was incorporated into the soil. The ratio between plant- and microbial-derived sugars together with delta13C values of individual sugars in the 2-7.5 cm soil depth revealed that the dissipation of sugars from the 0-2 cm soil depth was not only due to leaching, but also was caused by microbial degradation of the fresh C because slurry did not contain significant amounts of rhamnose while the delta13C values of rhamnose became progressively enriched in 13C during the experiment. Stable isotope measurements of bulk soil previously only showed significant differences between C4 and C3 plots at 0-2 cm soil depth. The CSIA of the individual sugars was much more sensitive than bulk isotope measurements, revealing significant differences between C4 and C3 plots even at the 2-7.5 cm soil depth during the first phase of the experiment. Additionally, the dynamics of slurry-derived plant and microbial sugars could be followed specifically.     

Influence of recent vegetation on labile and recalcitrant carbon soil pools in central Queensland, Australia: evidence from thermal analysis-quadrupole mass spectrometry-isotope ratio mass spectrometry

The effect of a recent vegetation change (<100 years) from C(4) grassland to C(3) woodland in central Queensland, Australia, on soil organic matter (SOM) composition and SOM dynamics has been investigated using a novel coupled thermogravimetry-differential scanning calorimetry-quadrupole.mass spectrometry-isotope ratio mass spectrometry (TG-DSC-QMS-IRMS) system. TG-DSC-QMS-IRMS distinguishes the C isotope composition of discrete SOM pools, showing changes in labile, recalcitrant and refractory carbon in the bulk soil and particle size fractions which track the vegetation changes. Analysis of evolved gases (by QMS) from thermal decomposition, rather than observed weight loss, proved essential in determining the temperature at which SOM decomposes, because smectite and kaolinite clays contribute to observed weight losses. The delta(13)C analyses of the CO(2) evolved at different temperatures for bulk soil and particle size-separates showed that most of the labile SOM under the more recent woody vegetation was C(3)-derived carbon whereas the delta(13)C values in the recalcitrant SOM showed greater C(4) contributions. This indicated a shift from grass (C(4))- to tree (C(3))-derived carbon in the woodland, which was also supported by the two-phase (13)C enrichment with depth, i.e. C(3) vegetation dominated the top soil (0-10 cm), but the C(4) contribution increased with depth (more gradual). This is perturbed by the inclusion of charcoal from forest fires ((14)C age incursions) and by the deep incorporation of C(3) carbon due to root penetration.     

Evolution of the delta13C signature related to total carbon contents and carbon decomposition rate constants in a soil profile under grassland

Evolution of the total carbon (C) content and the (13)C enrichment (delta(13)C signature) of soil organic matter (SOM) with increasing depth in a soil profile under permanent grassland (C(3) vegetation) were investigated. The relationship between the total C content and the delta(13)C signature at different depths in the upper 30 cm of the soil profile could be well fitted by the Rayleigh equation (y = -29.8 - 2.3x, R(2) = 0.95, p < 0.001), describing the enrichment in (13)C as resulting from isotopic fractionation associated with C mineralization (isotope enrichment factor epsilon = -2.3 per thousand). Potential C dynamics of SOM in four depth intervals of the profile (0-10, 10-20, 20-30 and 30-40 cm depth) were investigated through an incubation study. The C decomposition rate constants decreased with increasing sampling depth from 0.0479 yr(-1) (0-10 cm sampling depth) to 0.0256 yr(-1) (30-40 cm sampling depth) and were highly correlated (y = 0.02 + 0.13x, R(2) = 0.93, p < 0.05) with the corresponding deltadelta(13)C values (average change of the delta(13)C signature per depth increment). These results suggest that changes of the delta(13)C signature of SOM in undisturbed soil profiles under continuous C(3) vegetation may serve as an indicator of the variation of SOM quality with increasing depth.     

Enhancing the understanding of earthworm feeding behaviour via the use of fatty acid delta13C values determined by gas chromatography-combustion-isotope ratio mass spectrometry

Litter-dwelling (epigeic) Lumbricus rubellus and soil-dwelling (endogeic) Allolobophora chlorotica earthworms were observed aggregating under C(3) (delta(13)C = -31.3 per thousand; delta(15)N = 10.7 per thousand) and C(4) (delta(13)C = -12.6 per thousand; delta(15)N = 7.5 per thousand) synthetic dung pats applied to a temperate grassland (delta(13)C = -30.3 per thousand; delta(15)N = 5.7 per thousand) in an experiment carried out for 372 days. Bulk delta(13)C values of earthworms collected from beneath either C(3) or C(4) dung after 28, 56, 112 and 372 days demonstrated that (i) L. rubellus beneath C(4) dung were significantly (13)C-enriched after 56 days (delta(13)C = -23.8 per thousand) and 112 days (delta(13)C = -22.4 per thousand) compared with those from C(3) dung treatments (56 days, delta(13)C = -26.5 per thousand; 112 days, delta(13)C = -27.0 per thousand), and (ii) A. chlorotica were 2.1 per thousand (13)C-enriched (delta(13)C = -24.2 per thousand) relative to those from C(3) dung (delta(13)C = -26.3 per thousand) treatments after 372 days. Bulk delta(15)N values did not suggest significant uptake of dung N by either species beneath C(3) or C(4) dung, but showed that the endogeic species (total mean delta(15)N = 3.3 per thousand) had higher delta(15)N values than the epigeic species (total mean delta(15)N = 5.4 per thousand). Although the two species exhibited similar fatty acid profiles, individual fatty acid delta(13)C values revealed extensive routing of dietary C into body tissue of L. rubellus, but minor incorporation into A. chlorotica. In particular, the direct incorporation of microbial biomarker fatty acids (iC(17:0), aC(17:0)) from (13)C-labelled dung in situ, the routing of dung C into de novo synthesised compounds (iC(20:4)(omega)(6),C(20:5)(omega)(3), and the assimilation of essential fatty acids ((C(18:1)(omega)(9), C(18:1)(omega(7), C(18:2)(omega(6), C(18:3)(omega)(3)) derived from dung, were determined.     

Vertical gradients of delta15N and delta18O in soil atmospheric N2O--temporal dynamics in a sandy soil

The greenhouse gas nitrous oxide (N(2)O) can be both formed and consumed by microbial processes in the soil. As these processes fractionate strongly in favour of (14)N and (16)O, delta(15)N and delta(18)O gradients of N(2)O in the soil profile may elucidate patterns of N(2)O formation, consumption or emission to the atmosphere. We present the first in situ data of such gradients over time for a mesic typic Haplaquod seeded with potatoes (Solanum tuberosum L.). On two adjacent fields in 2002 and 2003, topsoil N(2)O fluxes were measured and the soil atmosphere was regularly sampled for N(2)O concentrations, delta(15)N and delta(18)O signatures of N(2)O at depths of 18, 48 and 90 cm during approximately 400 days. During the entire sampling period, the N(2)O concentrations were the highest and the delta(15)N signatures the lowest in the subsoil (48 or 90 cm depth) as compared with the topsoil, indicating production of N(2)O in the subsoil. For delta(15)N, differences greater than 30 per thousand between topsoil and subsoil on the same date were regularly observed. The highest N(2)O concentration of 100385 microL m(-3) at 90 cm depth on 1 July 2003, was preceded by the lowest delta(15)N value of -43.5 per thousand one week earlier. This was followed by a 150-day general decrease of N(2)O concentrations at 90 cm depth to 1723 microL m(-3) and a simultaneous enrichment of delta(15)N to +7.1 per thousand, mostly without a significant topsoil flux. There was a negative logarithmic relationship between N(2)O concentration at 90 cm depth and its delta(15)N signature. This relationship indicated a delta(15)N signature of -40 to -45 per thousand during the production of N(2)O in the subsoil, and a subsequent enrichment during the consumption of N(2)O. We conclude that the isotopic signature of the N(2)O topsoil flux is the result of various processes of consumption and production at different depths in the soil profile. It is therefore not a reliable estimator for the overall delta(15)N signature of N(2)O in the soil atmosphere, nor for indirect losses of N(2)O to the environment. Therefore, these findings will pose a further challenge to ongoing efforts to draw up a global isotopic budget for N(2)O.     

The effect of afforestation on the soil organic carbon (SOC) of a peaty gley soil using on-line thermally assisted hydrolysis and methylation (THM) in the presence of 13C-labelled tetramethylammonium hydroxide (TMAH)

In Britain substantial areas of both deep and shallow peatland have been afforested with conifers since the 1950s. However, information about the effects of afforestation on the properties of soil organic carbon (SOC) is lacking. Investigating the geochemical changes that take place when lignin- and tannin-derived phenols are degraded and incorporated into SOC will provide us with an insight into soil carbon dynamics at the molecular level. Here we compare the phenolic distributions in two different peaty gley soil profiles using on-line thermally assisted hydrolysis and methylation (THM) in the presence of both unlabelled and ¹³C-labelled tetramethylammonium hydroxide (TMAH). The two soil profiles were beneath respectively an unforested moorland (ML) and a second rotation Sitka spruce (Picea sitchensis) afforested moorland (SS), from Harwood (Northumberland, northeast England, UK). THM of these soils in the presence of ¹³C-labelled TMAH enabled us to assess the relative contributions of lignin, demethylated lignin, and non-lignin phenolics. The lignin phenolic distributions differed in both soil profiles reflecting changing source and decay dynamics within and between the sites. A progressive degradation of syringyl (S) and guaiacyl (G) phenolics was observed in the ML soil, compared with an increase of such components in the organic/mineral horizons of the SS soil. A significant tannin input was observed, particularly in the upper horizons of the SS soil. The S/G ratio gradually decreased with increasing burial in the ML soil, whilst a change in vegetation input and land preparation was recorded by this ratio in the SS soil. Overall, this suggests that afforestation influences the phenolic compositional profiles in these peaty gley soils as a result of one or more of the following processes: changing vegetation input, horizon inversion prior to planting, root input or leaching. This highlights the potential of using lignin and tannins as molecular indicators to assess the effects of afforestation on SOC.     

Applications of stable isotope ratio mass spectrometry in cattle dung carbon cycling studies

Understanding the fate of dung carbon (C) in soils is challenging due to the ubiquitous presence of the plant‐derived organic matter (OM), the source material from which both dung‐derived OM and soil organic matter (SOM) predominantly originate. A better understanding of the fate of specific components of this substantial source of OM, and thereby its contribution to C cycling in terrestrial ecosystems, can only be achieved through the use of labelled dung treatments. In this short review, we consider analytical approaches using bulk and compound‐specific stable carbon isotope analysis that have been utilised to explore the fate of dung‐derived C in soils. Bulk stable carbon isotope analyses are now used routinely to explore OM matter cycling in soils, and have shown that up to 20% of applied dung C may be incorporated into the surface soil horizons several weeks after application, with up to 8% remaining in the soil profile after one year. However, whole soil δ¹³C values represent the average of a wide range of organic components with varying δ¹³C values and mean residence times in soils. Several stable ¹³C isotope ratio mass spectrometric methods have been developed to qualify and quantify different fractions of OM in soils and other complex matrices. In particular, thermogravimetry‐differential scanning calorimetry‐isotope ratio mass spectrometry (TG‐DSC‐IRMS) and gas chromatography‐combustion‐IRMS (GC‐C‐IRMS) analyses have been applied to determine the incorporation and turnover of polymeric plant cell wall materials from C₄ dung into C₃ grassland soils using natural abundance ¹³C isotope labelling. Both approaches showed that fluxes of C derived from polysaccharides, i.e. as cellulose or monosaccharide components, were more similar to the behaviour of bulk dung C in soil than lignin. However, lignin and its 4‐hydroxypropanoid monomers were unexpectedly dynamic in soil. These findings provide further evidence for emerging themes in biogeochemical investigations of soil OM dynamics that challenge perceived concepts of recalcitrance of C pools in soils, which may have profound implications for the assessment of the potential of agricultural soils to influence terrestrial C sinks. Copyright © 2010 John Wiley & Sons, Ltd.     

Relationship between soil organic C degradability and the evolution of the delta13C signature in profiles under permanent grassland

The main objective of this research was to investigate to what extent the potential C dynamics of soil organic matter (SOM) are related to the degree of 13C enrichment with increasing depth in soil profiles under permanent grassland. The evolution of the C content and the 13C natural abundance (delta13C value) of SOM were investigated in three soil profiles (0-40 cm depth) under permanent grassland of varying texture (a loamy sand, a loam and a clay loam soil). The delta13C value of the SOM showed a gradual increase with increasing depth and decreasing C content in the profiles, ranging from 1.9 per thousand (loamy sand soil), 2.9 per thousand (clay loam soil) and 4 per thousand (loam soil) in relation to the delta13C value of SOM at the surface. The relationship between the 13C enrichment and total organic C content at different depths in the profiles (down to 40 cm depth in the loam and clay loam soil, down to 25 cm depth in the loamy sand soil) could be well described by the Rayleigh equation. The enrichment factors epsilon, associated with the Rayleigh approximation of the data, ranged from -1.57 per thousand (clay loam soil) to -1.64 per thousand (loamy sand soil) and -1.91 per thousand (loam soil). The potential C dynamics in four depth intervals from the profiles (0-10, 10-20, 20-30 and 30-40 cm depth) were determined by means of an incubation experiment. The C decomposition rate constants from the four sampling depths in the profiles showed a significant, positive correlation (y = 0.21x + 0.018, R(2) = 0.66, p < 0.005) with the corresponding Deltadelta13C values (change of the delta13C value per depth increment). A better correlation was obtained when only the data from the upper 20 cm in the profiles (y = 0.21x + 0.019, R(2) = 0.78, p < 0.05) were considered. These results suggest that the Deltadelta13C values in the surface layers of profiles under permanent grassland may serve as an indicator of the potential degradability or the stability of the SOM (in terms of C decomposition rate constants).     

Effects of dung and urine amendments on the isotopic content of N(2)O released from grasslands

The temporal and diurnal changes in nitrous oxide (N(2)O) fluxes were measured between 29(th) September and 2(nd) November 1999 from urine and dung patches from cattle deposited on grazed grassland. The delta(15)N and delta(18)O values of the N(2)O emitted from soil from both treatments were examined on four occasions during this period. The diurnal fluxes of N(2)O were measured by a chamber technique that provides hourly measurement of N(2)O fluxes. The (15)N and (18)O analysis of N(2)O were determined by isotope ratio mass spectrometry. N(2)O fluxes from the excreta patches were large, with peak emissions up to 1893 ng N m(-2) s(-1) occurring after heavy precipitation, measured one month after the treatment applications. Emissions from the urine patches were significantly greater than from the dung. The results showed that excretal patches are an important source of atmospheric N(2)O. The flux pattern showed a strong diurnal variation with maximum fluxes generally occurring in late afternoon or early morning, and generally not in phase with the soil temperature changes. The isotopic content of (15)N and (18)O in the N(2)O showed a similar trend to that of the N(2)O flux. The (15)N and (18)O values of the N(2)O emitted from the soil indicated that denitrification was the major process involved. After heavy precipitation on the 6(th) October, the larger delta(15)N and delta(18)O values suggested a consumption of the N(2)O by total denitrification.     

Application of Pyrolysis Cryogenic Trap Gas Chromatography/Atomic Emission Detection (Py-CryT-GC/AED) for the Study of Nitrogen Forms and Dynamics in a Grassland Soil

Soil organic nitrogen (N) was characterized for its chemical species and chemical transformations in a grassland soil profile by using a combination of cryogenic pyrolysis gas chromatography atomic emission detection method with soil physical size fractionation. The soils taken from 0–12, 12–25, and 25–38 cm depth layers were separated into five fractions, <2, 2–38, 38–53, 53–105, and 105–250 µm and each of which was analyzed for total organic C and N, and different N forms. Our results showed that (1) total organic carbon has a positive correlation with the total organic nitrogen (TON) with correlation coefficient increased with soil depth; (2) deep and small particle-size fraction soils yielded more volatile pyrolysate N than the surface and large particle-size fractions and the amount of volatile pyrolysate N has a linear positive correlation with TON and correlation coefficient increased with soil depth; (3) the major components of the volatile pyrolysate N include ammonia, acetonitrile, hydrogen cyanide, pyridine, and pyrrole; (4) of the total volatile pyrolysate N, ammonia accounted for more than 40%, and the sum of acetonitrile and hydrogen cyanide accounted for approximately 30–50%; and (5) the amounts of acetonitrile, hydrogen cyanide, and pyridine had increased positive correlations with TON with increasing soil depth, but the correlation between the amount of pyrrole and depth decreased in the opposite direction. Our research result sheds some light into soil organic nitrogen forms and its transformations in the processes of soil organic C aging and stabilization.     

δ15N natural abundance in permafrost soil indicates impact of fire on nitrogen cycle

The impact of fire on the nitrogen (N) cycle of natural ecosystems is arguable. Here we report and interpret an observation from boreal ecosystems in the Lena River basin, Sakha Republic (Yakutia), Russian Federation. Different types of permafrost soil (0-30 cm depth) were sampled along transects (60-150 m length) from the forest edge towards the centre of four separate thermokarst depressions under grassland. The average values of δ(15)N were remarkably similar within three transects, but differed systematically between them. Three findings point towards fire being the cause of the observed pattern. First, the spatial extent of systematic differences in soil δ(15)N coincides with the extent of typical fire scars in the region. Second, soil enrichment in (15)N is larger in the proximity of settlements, where fire is generally more frequent than in more remote places. Third, there is a significant positive correlation between δ(15)N values and the ratio of black C to total N. These findings point towards fire having a marked impact on soil δ(15)N and, accordingly, on the N cycle of this cold and dry ecosystem.     

High rates of nitrogen cycling in volcanic soils from Chilean grasslands

There are over one million hectares of pasture in Chile, and 80% and 50% of the country's milk and meat comes from 72% of this area, situated in the lake region of southern Chile. The soils are volcanic and a major characteristic is that they have very high organic matter (OM) contents with the potential to support plant growth with only moderate levels of added nitrogen (N). To understand better the potential fertility of these soils in order to maximise production and minimise losses of N, we undertook studies using the stable isotope of N ((15)N) to resolve the rates of the main internal N cycling processes in three soils representing the two main volcanic soil types: Osorno and Chiloé (Andisol) and Cudico (Ultisol). We also assessed the longer-term potential of these soils to sustain N release using anaerobic incubation. Gross rates (μg N g(-1) day(-1)) of mineralisation were 27.9, 27.1 and 15.5 and rates of immobilisation were 5.9, 12.0 and 6.3 for Osorno, Chiloé and Cudico, respectively, implying high rates of net mineralisation in these soils. This was confirmed by anaerobic incubation which gave potential seasonal net mineralisation indices of 1225, 1059 and 450 kg N ha(-1) in the top 10 cm soil layers of the three soils. However, plant production may still benefit from added N, as the release of N from organic sources may not be closely synchronised with crop demand. The low rates of nitrification that we found with these acidic soils suggest that the more mobile N (viz. nitrate-N) would be in limited supply and plants would have to compete for the less mobile ammonium-N with the soil microbial biomass. Nitrogen was mineralised in appreciable amounts even down to 60 cm depth, so that leaching could become significant, particularly if the soils were limed, which could enhance nitrification and N mobility through the soil profile.     

Off-line pyrolysis and compound-specific stable carbon isotope analysis of lignin moieties: a new method for determining the fate of lignin residues in soil

Off-line pyrolysis was used to liberate lignin moieties from dung and soil and, after trimethylsilylation, the delta(13)C values of these derivatives were determined by gas chromatography-combustion-isotope ratio mass spectrometry. Initial delta(13)C values determined for 4-vinylphenol, syringol, 4-vinylguaiacol, 4-acetylsyringol, 4-vinylsyringol, 4-(2-Z-propenyl)syringol, 4-(2-E-propenyl)syringol and 4-(2-propenone)syringol pyrolysis products of the lignin polyphenol structure from C(4) (delta(13)C(bulk) = -12.6%) and C(3) (delta(13)C(bulk) = -30.1 per thousand) dung confirmed the robust and reproducible nature of the off-line preparation technique. C(4) dung was used as a treatment in a randomised field experiment to assess the short-term sequestration of dung carbon in managed grasslands. Since lignin was on average 3.5 per thousand depleted in (13)C compared with bulk dung delta(13)C values, this may have resulted in an under-estimation of dung C incorporation based on bulk delta(13)C values. Therefore, an investigation of the compound-specific delta(13)C values of dung-derived lignin moieties extracted from soils sampled up to 372 days was undertaken. Delta(13)C values between lignin moieties extracted from treated and untreated soils showed that dung-derived lignin was not especially resistant to degradation and suggested that individual moieties of the lignin macromolecule must: (i) move into soil, (ii) be degraded, or (iii) be transformed diagenetically at different rates. This adds to a gathering body of evidence that lignin is not particularly stable in soils, which has considerable significance for the perceived role of different biochemical components in the cycling of C in soils.     

Oxalate distribution in soils under rhubarb (Rheum rhaponticum)

Oxalate in soils may enhance phosphate availability, promote mineral dissolution, and increase the mobility of aluminium and heavy metal cations by complexation. Rhubarb (Rheum rhaponticum L.) has very high content of oxalate in leaves and petioles, and therefore the topsoil under rhubarb might have elevated contents of oxalate. Soil samples were collected at depths of 0–2.5 and 2.5–5?cm from 10?cm sections along 100?cm transects from rhubarb plants at four locations in Denmark, and from seven layers in a soil profile to 80?cm depth at one location. Oxalate was extracted from the soil with 0.2?M phosphate at pH 2 by reciprocal shaking for 24?h and then determined by a new fast capillary zone electrophoresis method with 300?mM KH₂PO₄ and 0.30?mM TTAB electrolyte adjusted to pH 7, developed and tested to analyse high-ionic-strength soil extracts. Rhubarb increases the oxalate content in soil under the leaves slightly. The average content of oxalate in the upper 0–5?cm soil was 444?µmol/kg at the Kaldred site, and 111–333?µmol/kg at the three other locations. In the soil profile, the content of oxalate decreased from 500?µmol/kg in 0–5?cm depth to 110?µmol/kg at 75–80?cm depth. No significant seasonal changes in oxalate contents were observed, while an annual variation of 100?µmol/kg could be observed at 0–2.5?cm depth. During plant decay in autumn, a slight increase in oxalate content was observed at 30?cm soil depth. In conclusion, the role of oxalate in weathering and metal transport appears to be limited in soils under rhubarb. Oxalate might stimulate microbiological growth and phosphate mobilisation in the rhizosphere, but concentrations observed are too low to impose any toxic effects to organisms.     

Long-term influence of manure and mineral nitrogen applications on plant and soil 15N and 13C values from the Broadbalk Wheat Experiment

The Broadbalk Wheat Experiment at Rothamsted Research in the UK provides a unique opportunity to investigate the long-term impacts of environmental change and agronomic practices on plants and soils. We examined the influence of manure and mineral fertiliser applications on temporal trends in the stable N ((15)N) and C ((13)C) isotopes of wheat collected during 1968-1979 and 1996-2005, and of soil collected in 1966 and 2000. The soil delta(15)N values in 1966 and 2000 were higher in manure than the mineral N supplied soil; the latter had similar or higher delta(15)N values than non-fertilised soil. The straw delta(15)N values significantly decreased in all N treatments during 1968 to 1979, but not for 1996-2005. The straw delta(15)N values decreased under the highest mineral N supply (192 kg N ha(-1) year(-1)) by 3 per thousand from 1968 to 1979. Mineral N supply significantly increased to straw delta(13)C values in dry years, but not in wet years. Significant correlations existed between wheat straw delta(13)C values with cumulative rainfall (March to June). The cultivar Hereward (grown 1996-2005) was less affected by changes in environmental conditions (i.e. water stress and fertiliser regime) than Cappelle Desprez (1968-1979). We conclude that, in addition to fertiliser type and application rates, water stress and, importantly, plant variety influenced plant delta(13)C and delta(15)N values. Hence, water stress and differential variety response should be considered in plant studies using plant delta(13)C and delta(15)N trends to delineate past or recent environmental or agronomic changes.     

Evaluation of methods to measure differential 15N labeling of soil and root N pools for studies of root exudation

To study patterns of root exudation, the effectiveness of different techniques for in situ 15N labeling of Brassica napus, Centaurea jacea and Lolium perenne with ammonium nitrate was tested. Stem infiltration was found to effectively label plants with thicker stems, whereas, for grass species, cutting and immersing the leaf tips into 15N solution proved to be most effective. A microdiffusion technique to isolate ammonium, combined with conventional cation-exchange chromatography to separate nitrate from amino-N compounds thereafter, was found suitable for separation of the N fractions of plant and soil extracts for 15N determination. All three species were then cultivated in nutrient solution and labeled with 15NH4 15NO3 by stem feeding for 42 hours. Kinetics of 15N labeling of bulk roots and shoots as well as hot water extractable material were assessed, and up to 1.1 at% 15N excess (APE) was found in nutrient solutions. The main amino acids exuded by L. perenne were glycine, serine, alanine and aspartic acid. To assess the suitability of this set of methods to study root exudation in field settings, L. perenne was grown without fertiliser addition in pots containing low-nutrient soil. Plants were 15N labeled via tip immersion and 15N and N concentrations were analysed in shoots, roots and soils during a 48-h interval. Shoots reached 1.25 APE, roots and soil 0.10 and 0.005 APE, respectively. Between 4% (48 h) and 6% (24 h) of total plant 15N was exuded by roots into the soil. In roots amino acids comprised the largest proportion of the soluble 15N pool, whereas soil 15N levels were similar for amino acids and ammonium, exceeding those of nitrate. Mechanisms for the shift within N fractions from roots to soils are briefly discussed.     

Belowground fate of 15N injected into sweetgum trees (Liquidambar styraciflua) at the ORNL FACE Experiment

Nitrogen (N) cycling can be an important constraint on forest ecosystem response to elevated atmospheric CO₂. Our objective was to trace the movement of ¹⁵N, injected into tree sap, to labile and stable forms of soil organic matter derived partly from the turnover of tree roots under elevated (545 ppm) and ambient (394 ppm) atmospheric CO₂ concentrations at the Oak Ridge National Laboratory (ORNL) FACE (Free‐Air Carbon Dioxide Enrichment) Experiment. Twenty‐four sweetgum trees, divided equally between CO₂ treatments, were injected with 3.2 g ¹⁵N‐ammonium sulfate (99 atom %), and soil samples were collected beneath the trees over a period of 89 weeks. For 16 cm deep soil samples collected beneath the study trees, there was 28% more fine root (less than or equal to 2 mm diameter) biomass under elevated CO₂ (P = 0.001), but no significant treatment effect on the amounts of necromass, coarse root biomass, or on the N concentrations in tree roots and necromass. Nitrogen‐15 moved quickly into roots from the stem injection site and the ¹⁵N content of roots, necromass, and labile organic matter (i.e. particulate organic matter, POM) increased over time. At 89 weeks post‐injection, approximately 76% of the necromass ¹⁵N originated from fine root turnover. Nitrogen‐15 in POM had a relatively long turnover time (47 weeks) compared with ¹⁵N in roots (16 to 22 weeks). Over the 1.7 year period of the study, ¹⁵N moved from roots into slower cycling POM and the disparity in turnover times between root N and N in POM could impose progressive limitations on soil N availability with stand maturation irrespective of atmospheric CO₂, especially if the release of N through the decomposition of POM is essential to sustain forest net primary production. Published in 2009 by John Wiley & Sons, Ltd.     

Quantification of priming and CO2 respiration sources following slurry-C incorporation into two grassland soils with different C content

The fate of incorporated slurry-C was examined in a laboratory experiment using two UK grassland soils, i.e. a Pelostagnogley (5.1 %C) and a Brown Earth (2.3 %C). C3 and C4 slurries were incorporated into these two wet-sieved (C3) soils (from 4-10 cm depth). Gas samples were collected 0.2, 1, 2, 3, 4, 6, 9, 20, 30 and 40 days after slurry application and analyzed for CO2 concentration and delta13C content. Slurry incorporation into the soil strongly increased soil CO2 respiration compared with the unamended soil. Total (40 day) cumulative CO2 flux was higher for the Pelostagnogley than the Brown Earth. The 13C natural abundance tracer technique enabled quantification of the sources of respired CO2 and priming effects (days 0-9). Proportionally more slurry-derived C was respired from the Pelostagnogley (46%) than the Brown Earth (36%). The incorporated slurry-C was lost twice as fast as the native soil C in both soils. Slurry incorporation induced a priming effect, i.e. additional release of soil-derived C, most pronounced in the Pelostagnogley (highest C content). The majority of respired soil-derived C (>70%) was primed C. The study indicated that potential reductions in ammonia volatilisation following slurry injection to grasslands might be negated by enhanced loss of primed soil C (i.e. pollution swapping).     

Quantifying nitrate retention processes in a riparian buffer zone using the natural abundance of 15N in NO3-

Quantifying the relative importance of denitrification and plant uptake to groundwater nitrate retention in riparian zones may lead to methods optimising the construction of riparian zones for water pollution control. The natural abundance of 15N in NO3- has been shown to be an interesting tool for providing insights into the NO3- retention processes occurring in riparian zones. In this study, 15N isotope fractionation (variation in delta15N of the residual NO3-) due to denitrification and due to plant uptake was measured in anaerobic soil slurries at different temperatures (5, 10 and 15 degrees C) and in hydroponic systems with different plant species (Lolium perenne L., Urtica dioica L. and Epilobium hirsutum L.). It was found that temperature had no significant effect on isotope fractionation during denitrification, which resulted in a 15N enrichment factor epsilonD of -22.5 +/- 0.6 per thousand. On the other hand, nitrate uptake by plants resulted in 15N isotope fractionation, but was independent of plant species, leading to a 15N enrichment factor epsilonP of -4.4 +/- 0.3 per thousand. By relating these two laboratory-defined enrichment factors to a field enrichment factor for groundwater nitrate retention during the growing season (epsilonR = -15.5 +/- 1.0 per thousand ), the contribution of denitrification and plant uptake to groundwater nitrate retention could be calculated. The relative importance of denitrification and plant uptake to groundwater nitrate retention in the riparian buffer zone was 49 and 51% during spring, 53 and 47% during summer, and 75 and 25% during autumn. During wintertime, high micropore dissolved organic carbon (DOC) concentrations and low redox potentials due to decomposition of the highly productive riparian vegetation probably resulted in a higher denitrification rate and favoured other nitrate retention processes such as nitrate immobilisation or dissimilatory nitrate reduction to ammonium (DNRA). This could have biased the 15N isotope fractionation and led to a low 15N enrichment factor for groundwater nitrate retention during wintertime (-6.2 +/- 0.9 per thousand ). In contradiction to what many other studies suggest, it is possible that due to plant decomposition during the winter period other nitrate transformation processes compete with denitrification.     

Forms of soil organic nitrogen in a Pactola soil as seen by pyrolysis/gas chromatography/atomic emission detection (Py–GC/AED)

In this study, soil organic nitrogen (SON) forms from a Pactola forest soil were investigated by using pyrolysis–cryogenic gas chromatography/atomic emission (Py–GC/AED). The samples were taken at different soil depths of 0–12 cm, 12–25 cm and 25–38 cm and separated into particle-size fractions. Each fraction was analyzed by Py–GC/AED at 300, 400 and 500 °C consecutively. The main N-structures found in pyrolysates were ammonia, acetonitrile, hydrogen cyanide, pyridine and pyrrole. Ratios of acetonitrile and hydrogen cyanide to pyridine and pyrrole decreased with soil depth. Acetonitrile and hydrogen cyanide had positive correlations with total organic N (TON) down the soil profile. A decrease of the optimal pyrolysis temperatures of soil samples with increasing soil depth was observed for acetonitrile and hydrogen cyanide. In the surface soil samples, there was no correlation found for both pyridine and pyrrole with TON. But with depth, the correlation coefficient increased and reached to 0.912 for pyridine, and 0.875 for pyrrole at the depth of 25–38 cm. The increased correlation coefficients of pyridine and pyrrole with TON in combination with the low shifts of the optimal pyrolysis temperatures of the soil samples down the soil profile for acetonitrile and hydrogen cyanide indicated clear differences in N-structures between different soil layers.