Nitrogen isotope analysis of free glycine in soil using isotope dilution mass spectrometry

To enable the estimation of production and consumption rates of free glycine in soils through ¹⁵N isotope dilution experiments, an isotope dilution mass spectrometric method was developed. The method, which enabled high precision N isotope ratio determination of glycine in soil extracts at δ¹⁵N levels up to 4000‰ and concentrations from approximately 2 μM, is based on the following steps: (i) addition of glycine spike to the soil extract, (ii) removal of humic substances and pre-concentration of glycine using solid phase extraction, (iii) derivatization of amino acids, (iv) separation of the derivatives using gas chromatography (GC), (v) their combustion to yield sample N₂ gas, and (vi) finally the use of N isotope ratio mass spectrometry (IRMS). Judging by uncertainty budget calculations, the precision obtained (SD=0.01-0.06 at% ¹⁵N) is sufficient for detecting differences in N isotopic ratios obtained in ¹⁵N isotope dilution experiments.     

Compound‐specific stable‐isotope (δ13C) analysis in soil science

This review provides current state of the art of compound‐specific stable‐isotope‐ratio mass spectrometry (δ¹³C) and gives an overview on innovative applications in soil science. After a short introduction on the background of stable C isotopes and their ecological significance, different techniques for compound‐specific stable‐isotope analysis are compared. Analogous to the δ¹³C analysis in bulk samples, by means of elemental analyzer–isotope‐ratio mass spectrometry, physical fractions such as particle‐size fractions, soil microbial biomass, and water‐soluble organic C can be analyzed. The main focus of this review is, however, to discuss the isotope composition of chemical fractions (so‐called molecular markers) indicating plant‐ (pentoses, long‐chain n‐alkanes, lignin phenols) and microbial‐derived residues (phospholipid fatty acids, hexoses, amino sugars, and short‐chain n‐alkanes) as well as other interesting soil constituents such as “black carbon” and polycyclic aromatic hydrocarbons. For this purpose, innovative techniques such as pyrolysis–gas chromatography–combustion–isotope‐ratio mass spectrometry, gas chromatography–combustion–isotope‐ratio mass spectrometry, or liquid chromatography–combustion–isotope‐ratio mass spectrometry were compared. These techniques can be used in general for two purposes, (1) to quantify sequestration and turnover of specific organic compounds in the environment and (2) to trace the origin of organic substances. Turnover times of physical (sand < silt < clay) and chemical fractions (lignin < phospholipid fatty acids < amino sugars ≈ sugars) are generally shorter compared to bulk soil and increase in the order given in brackets. Tracing the origin of organic compounds such as polycyclic aromatic hydrocarbons is difficult when more than two sources are involved and isotope difference of different sources is small. Therefore, this application is preferentially used when natural (e.g., C3‐to‐C4 plant conversion) or artificial (positive or negative) ¹³C labeling is used.     

Measurement of nitrous oxide emissions from grassland soil using photo‐acoustic infra‐red spectroscopy,long‐path infra‐red spectroscopy,gas chromatography,and continuous flow isotope‐ratio mass spectrometry

Abstract

This study evaluated the performance of photo‐acoustic infra‐red spectroscopy (PAIRS) for measuring nitrous oxide (N₂O) fluxes in the field, in comparison with long‐path infra‐red spectroscopy ('Hawk'), gas chromatography (GC), and continuous flow isotope‐ratio mass spectrometry (CF‐IRMS). The N₂O flux measurements from fertilized and grazed grassland were made simultaneously by the different methods, before and after water application. Before irrigation, mean N₂O fluxes ranged from 3 to 20 g N ha¹ day¹ for the PAIRS and GC measurements, but were undetectable with the Hawk. Within 2 hours of irrigation, mean fluxes increased to 740, 640, and 270 g N ha‘¹day¹, based on GC, PAIRS, and Hawk measurements, respectively. After about 24 hours, irrigation had reached its full effect and N₂O fluxes had increased to 1,050,710, and 410 g N ha¹ day¹. The GC measurements were consistently higher than the PAIRS measurements. However, a second experiment, comparing the PAIRS analyzer with continuous flow isotope‐ratio mass spectrometry (CF‐IRMS), suggested that the former was negatively biased; the PAIRS response was about 33% lower than CF‐IRMS. When the resulting correction factor was applied to the results of the first experiment, there was very good agreement between the PAIRS and GC measurements. The Hawk measurements were lower than PAIRS and GC, but a statistical comparison was not possible, due to the limited number of Hawk measurements that could be made in the windy weather conditions. Windy conditions also resulted in an underestimation of the N₂O flux by PAIRS compared to GC and CF‐IRMS analysis, which could not solely be attributed to a change in the analyzer sensitivity. There was no obvious explanation for this discrepancy and further investigations are needed to resolve this issue.     

Amino acid 15N in long-term bare fallow soils: influence of annual N fertilizer and manure applications

Long‐term dynamics of amino acids (AAs), from a bare fallow soil experiment (established in 1928 at INRA‐Versailles, France), were examined in unamended control (Con) plots and plots treated with ammonium sulphate (Amsul), ammonium nitrate (Amnit), sodium nitrate (Nanit) or with animal manure (Man). Topsoil (0–25 cm) from 1929, 1963 and 1997 was analysed for C, N and ¹⁵N content and distribution of 18 amino acids recovered after acid hydrolysis with 6 m HCl. With time, soil N, C and AA content were reduced in Con, Amsul, Amnit and Nanit, but increased in Man. However, the absolute N loss was 3–11 times larger in Man than Nanit, Amsul, Amnit and Con, due to the much higher N annual inputs applied to Man. From 1929 to 1997 in Con, Amsul, Amnit and Nanit the whole soil and non‐hydrolysable‐N pool δ¹⁵N increased associated with the loss of N (indicative of Rayleigh ¹⁵N^/14N fractionation). No δ¹⁵N change from 1929 to 1997 was found in the hydrolysable AA‐N (HAN) pool. Fertilizer N inputs aided stabilization of soil AA‐N, as AA half‐life in the mineral N fertilizer treatments increased from 34 years in 1963 to 50 years in 1997. The δ¹⁵N values of alanine and leucine reflected both source input and ¹⁵N^/14N fractionation effects in soils. The δ¹⁵N increase of ornithine (～6‰) was similar to the whole soil. The δ¹⁵N change of phenylalanine in Con (decrease of 7‰) was related to its proportional loss since 1929, whereas for Amsul, Amnit, Nanit and Man it was associated with isotope effects caused by the fertilizer inputs. However, the soil δ¹⁵N value of most individual amino acids (IAAs) did not significantly change over nearly 70 years, even with mineral or organic N inputs. We conclude for these bare fallow systems that: (i) δ¹⁵N changes in the whole soil and non‐hydrolysable AA pool were solely driven by microbial processes and not by the nature of fertilizer inputs, and (ii) without plant inputs, the δ¹⁵N of the HAN pool and (most) IAAs may reflect the influence of plant–soil interactions from the previous (arable cropping) rather than present (fallow) land use on these soil δ¹⁵N values.     

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.     

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.     

Repeated annual drought has minor long‐term influence on δ13C and alkane composition of plant and soil in model grassland and heathland ecosystems

As a result of global climate change the incidence of drought conditions in Europe is predicted to increase in the future, which also influences plant resistance. Lipids are important plant constituents that protect plants against drought stress and contribute to the intermediate stable carbon (C) pool in soil. However, the extent to which drought influences lipid cycling in the plant–soil system is unknown and, therefore, it remains questionable how the ecosystem recovers after drought. We focused on plant and soil samples from two different plant communities (temperate grassland and heathland) that had been exposed to 5 years of 4.5–6.0 weeks repeated annual drought. They were sampled one year after the last drought to check the recovery of the plant–soil system. Samples were analyzed for their bulk C, stable C and nitrogen (N) isotope (δ¹³C, δ¹⁵N) and lipid composition. Contrary to our expectation, no strong influence of five years of repeated annual drought was observed for above‐ground biomass, roots and soils in the model ecosystems with respect to elemental (C and N concentrations, C : N ratio) bulk isotope (δ¹³C, δ¹⁵N) composition and the total extractable lipid concentration. Thus, plants did not sustain a significant change in their C and lipid concentration as well as their composition after five years of repeated annual drought. This might be related to the comparatively short drought period related to the overall growth season and provides evidence for recovery of the C and lipid dynamics in temperate grassland and heathland model ecosystems exposed to annual drought.     

Nitrogen isotope fractionation during nitrogen transport from ectomycorrhizal fungi,Suillus granulatus,to the host plant,Pinus densiflora

The ¹⁵N natural abundance in Pinus densiflora Sieb. et Zucco that had been inoculated and not inoculated with ectomycorrhizal fungi (Suillus granulatus (L.:Fr.) O. Kuntze) was compared. The inoculated pine needles showed a lower δ¹⁵N value, while the uninoculated ones showed a higher δ¹⁵N value. Higher δ¹⁵N values in the mycelial mat of the ectomycorrhizal fungi compared to those of the inoculated pine needles were also observed. These facts indicate that nitrogen isotope fractionation occurred during the nitrogen transport from mycorrhizal fungi to the host plants.     

In-source pyrolysis-field ionization mass spectrometry and Curie-point pyrolysis-gas chromatography/mass spectrometry of amino acids in humic substances and soils

With the aid of in-source pyrolysis-field ionization mass spectrometry (Py-FIMS) and Curie-point pyrolysis-gas chromatography/mass spectrometry (cPy-GC/MS) in the conventional electron impact mode, characteristic signals of 23 amino acid standards were described. Thermal and mass spectrometric fragmentation pathways of these amino acids differed with each method and complemented each other. Pyrolysis products assigned by Py-FIMS extended the range of signals for N-containing compounds in humic substances and soil organic matter and gave marker signals for free amino acids and their subunits in proteinaceous materials. These characteristic signals were correlated with the amino acid content in N-rich humic fractions consisting of seven fulvic acids and eight humic acids. The selected marker signals reflected 25–84% of the variances of the molar distribution of acidic, neutral, neutral aromatic, and basic amino acids in the humic fractions. In addition, a well described agricultural soil (0.08% amino acid N) was spiked with a standard amino acid mixture (0.08 mg amino acid N 100 mg^-1 dry soil) and produced enhancements of the relative abundances of the corresponding amino acid signals. Moreover, for 27 samples of whole agricultural soils of widely different origins, soil types, and vegetations, 15 selected amino acid indicators were correlated significantly with -amino N (r=0.76^***) and total N (r=0.65^***).     

Short‐term sequestration of slurry‐derived carbon and nitrogen in temperate grassland soil as assessed by 13C and 15N natural abundance measurements

Land application of animal wastes from intensive grassland farming has caused growing environmental problems during the last decade. This study aimed to elucidate the short‐term sequestration of slurry‐derived C and N in a temperate grassland soil (Southwest England) using natural abundance ¹³C and ¹⁵N stable isotope techniques. Slurry was collected from cows fed either on perennial ryegrass (C3) or maize (C4) silages. 50 m³ ha^—1 of each of the obtained C3 or C4 slurries (δ¹³C = —30.7 and —21.3‰, δ¹⁵N = +12.2 and + 13.8 ‰, respectively) were applied to a C₃ soil with δ¹³C and δ¹⁵N values of —30.0 ± 0.2‰ and + 4.9 ± 0.3‰, respectively. Triplicate soil samples were taken from 0—2, 2—7.5, and 7.5—15 cm soil depth 90 and 10 days before, at 2 and 12 h, as well as at 1, 2, 4, 7, and 14 days after slurry application and analyzed for total C, N, δ¹³C, and δ¹⁵N. No significant differences in soil C and N content were observed following slurry application using conventional C and N analysis techniques. However, natural abundance ¹³C and ¹⁵N isotope analysis allowed for a sensitive temporal quantification of the slurry‐derived C and N sequestration in the grassland soil. Our results showed that within 12 hours more than one‐third of the applied slurry C was found in the uppermost soil layer (0—2 cm), decreasing to 18% after 2 days, but subsequently increasing to 36% after 2 weeks. The tentative estimate of slurry‐derived N in the soil suggested a decrease from 50% 2 hours after slurry application to only 26% after 2 weeks, assuming that the increase in δ¹⁵N of the slurry plots compared to the control is proportional to the amount of slurry‐incorporated N. We conclude that the natural abundance tracer technique can provide a rapid new clue to the fate of slurry in agricultural C and N budgets, which is important for environmental impacts, farm waste management, and climate change studies.     

Spatially tracking carbon through the root–rhizosphere–soil system using laser ablation‐IRMS

The intimate relationships between plant roots, rhizosphere, and soil are fostered by the release of organic compounds from the plant into soil through various forms of rhizodeposition and the simultaneous harvesting of nutrients from the soil to the plant. Here we present a method to spatially track and map the migration of plant‐derived carbon (C) through roots into the rhizosphere and surrounding soil using laser ablation‐isotope ratio mass spectrometry (LA‐IRMS). We used switchgrass microcosms containing soil from field plots at the Kellogg Biological Station (Hickory Corners, Michigan, USA) which have been cropped with switchgrass since 2008. We used a ¹³CO₂ tracer to isotopically label switchgrass plants for two diel cycles and tracked subsequent movement of labeled C using the spatially specific (< 100 µm resolution) δ¹³C analysis enabled by LA‐IRMS. This approach permitted assessment of variable C flow through different roots and enabled mapping of spatial variability of C allocation to the rhizosphere. Highly ¹³C‐enriched C (consistent with production during the ¹³CO₂ application period) extended ≈ 0.5–1 mm from the root into the soil, suggesting that the majority of recent plant‐derived C was within this distance of the root after 48 h. Tracking the physical extent of root exudation into the rhizosphere can help evaluate the localization of plant‐microbe interactions in highly variable subsurface environments, and the use of the isotopic label can differentiate freshly fixed C (presumably from root exudates) from other types of subsurface C (e.g., plant necromass and microbial turnover). The LA‐IRMS technique may also serve as a valuable screening technique to identify areas of high activity for additional microbial or geochemical assays.     

Comparison of the difference and delta 15Nitrogen approaches for evaluating liquid urea ammonium nitrate utilization by maize

Abstract

Isotopic nitrogen (N) research techniques may be required in watershed studies to determine the impact of landscape position on fertilizer efficiency and the soil supplying power. However, traditional approaches using ^I5N labeled fertilizer may not be suitable when farmer equipment is used. The δ¹⁵N natural abundance isotopic approach has been used to evaluate N cycling in watersheds. The objectives of this study were to measure the precision of the δ¹⁵N measurement by the Europa 20–20 ratio mass spectrometer (Europa Scientific Ltd, UK), and to compare the difference and δ¹⁵N approaches for measuring fertilizer use by maize (Zea mays). A replicated field study containing two different N rates (0 and 15.7 g N m^‐2) were used for the study. Maize samples were collected at the 8th‐leaf, silking, and plant maturity in 1992 and 1993. Samples were dried (80°C), ground (<1‐mm), weighed (stover 12 mg and grain 3 mg), and analyzed fortotal N and δ¹⁵N. Fertilizer utilization at the three growth stages was determined using the natural abundance δ¹⁵N and nonisotopic difference (fertilizer‐control) techniques. During the study, the Europa 20–20 ratio mass spectrometer (Europa Scientific Ltd, UK) analyzed over a 100 samples a day and had consumable costs of less than $2.00 per sample. The standard deviations of the mean were less than 0.11 and 0.21 %o in 51 and 77% of the stover samples, respectively. In 1992, grain yields were not influenced by N fertilizer additions, while in 1993 grain yields were increased by N fertilizer. The difference method estimated that in 1992, 16% of the N fertilizer was utilized by the crop, while the natural abundance δ¹⁵N approach estimated that 36% of the fertilizer N was used by the crop. Differences between calculated values by the two techniques resulted from the difference method calculating net fertilizer use, while the δ¹⁵N approach calculated fertilizer contained in the plant. Because the δ¹⁵N approach estimates fertilizer use, this approach can be used to calculate soil N contained in the plant. In watershed studies, this information may provide the tools needed to evaluate N use in responsive and nonresponsive sites within a field. This research shows that the δ¹⁵N method compliments the difference method, can be used to measure actual fertilizer use when farmer equipment is used, and that the Europa 20–20 ratio mass spectro‐meter (Europa Scientific Ltd, UK) has acceptable precision for the δ¹⁵N natural abundance approach.     

Preparation of Ammonium‐15N and Nitrate‐15N Samples by Microdiffusion for Isotope Ratio Analysis by Optical Emission Spectrometry

Abstract

A diffusion method for the preparation and measurement of ¹⁵N abundance of ammonium and nitrate in KCl extracts of soil using optical emission spectrometry (OES) was compared with conventional continuous flow isotope ratio mass spectrometry (IRMS). There were highly significant correlations between the values obtained by using OES and IRMS. The 99% confidence interval of the intercept included the value 0 and the 99% confidence interval of the slope included the value 1 for both nitrate and ammonium measurements, suggesting that the results from the two methods did not differ significantly. In another experiment, ¹⁵N values of nitrate and ammonium from soil extracts prepared by using the standard distillation procedures for OES were compared against the microdiffusion preparation method. Again, there was a highly significant correlation between the values: the 95% confidence interval of the intercept included the value 0, and the 95% confidence interval of the slope included the value 1, again suggesting that the two methods did not differ significantly. It was concluded that the diffusion technique is an appropriate and simple method of sample preparation for inorganic N analysis of KCl extracts using OES.     

Responses of herbage and cattle tail switch hair δ15N value to long-term stocking rates on a rough fescue grassland

The purpose of this study was to investigate the response of δ¹⁵N in herbage and cattle tail switch hair to long-term grazing pressure on a rough fescue grassland (Festuca campestris Rydb.) near Stavely, Alberta, Canada. Cattle have grazed the paddocks from 15 May to 15 November annually since 1949. Stocking rates were 0, 2.4 and 4.8 animal unit months ha^?1 for non-grazing (Control), moderate grazing (MG) and heavy grazing (HG), respectively. Green standing crop (GSC) was sampled monthly throughout the grazing season in 2007. The GSC was fractioned into neutral detergent fiber (NDF), acid detergent fiber (ADF), and their total nitrogen (TN) concentration and δ¹⁵N values in NDF, ADF and GSC were determined. Tail switch hair samples from cows (>2 years old) and calves (<1 year) were collected at the end of the grazing season in 2007 and 2008 and analysed for δ¹⁵N values. The TN concentrations in NDF and δ¹⁵N values in herbage NDF and ADF fractions were higher (P?15N values in tail hair also decreased (P?15N values in tail hair increased with herbage δ¹⁵N values. The δ¹⁵N values in tail hair were enriched by +5.2‰ compared to herbage δ¹⁵N values in 2007. Changes in δ¹⁵N value in GSC and cattle hair reflect the influence of grazing practices on N cycles through the animal/plant/soil system on this rough fescue grassland.     

Do plant species with different growth strategies vary in their ability to compete with soil microbes for chemical forms of nitrogen?

We used dual labelled stable isotope (¹³C and ¹⁵N) techniques to examine how grassland plant species with different growth strategies vary in their ability to compete with soil microbes for different chemical forms of nitrogen (N), both inorganic and organic. We also tested whether some plant species might avoid competition by preferentially using different chemical forms of N than microbes. This was tested in a pot experiment where monocultures of five co-existing grassland species, namely the grasses Agrostis capillaris, Anthoxanthum odoratum, Nardus stricta, Deschampsia flexuosa and the herb Rumex acetosella, were grown in field soil from an acid semi-natural temperate grassland. Our data show that grassland plant species with different growth strategies are able to compete effectively with soil microbes for most N forms presented to them, including inorganic N and amino acids of varying complexity. Contrary to what has been found in strongly N limited ecosystems, we did not detect any differential uptake of N on the basis of chemical form, other than that shoot tissue of fast-growing plant species was more enriched in ¹⁵N from ammonium-nitrate and glycine, than from more complex amino acids. Shoot tissue of slow-growing species was equally enriched in ¹⁵N from all these N forms. However, all species tested, least preferred the most complex amino acid phenylalanine, which was preferentially used by soil microbes. We also found that while fast-growing plants took up more of the added N forms than slow-growing species, this variation was not related to differences in the ability of plants to compete with microbes for N forms, as hypothesised. On the contrary, we detected no difference in microbial biomass or microbial uptake of ¹⁵N between fast and slow-growing plant species, suggesting that plant traits that regulate nutrient capture, as opposed to plant species-specific interactions with soil microbes, are the main factor controlling variation in uptake of N by grassland plant species. Overall, our data provide insights into the interactions between plants and soil microbes that influence plant nitrogen use in grassland ecosystems.     

The influence of trophic level as measured by δ15N on mercury concentrations in freshwater organisms

The relationship between mercury (Hg) concentrations in freshwater biota and trophic position, as defined by stable nitrogen isotope ratios (δ¹⁵N), was examined in 6 lakes in northwestern Ontario. The heavier isotope of nitrogen (¹⁵N) increases an average of 3 parts per thousand (‰) from prey to predator and is used as a measure of an organism's trophic position. Dorsal muscle from lake trout, burbot, walleye, northern pike, white sucker, lake cisco, lake whitefish, and yellow perch was analyzed for Hg and δ¹⁵N using flameless atomic absorption and mass spectrometry respectively. Within each lake, log Hg was significantly related to δ¹⁵N (r ² ranged from 0.47 to 0.91,P<0.01). For four species, yellow perch, northern pike, lake cisco, and lake trout, log Hg was positively related to δ¹⁵N (r ² ranged from 0.37 to 0.47,P≤0.09) across all lakes. We also used δ¹⁵N measurements (assuming a 3‰ shift between an organism and its diet) and the developed within-lake regression equations to calculate a prey Hg for each individual fish. These food Hg values were then used to predict predator Hg using Norstromet al's bioenergetics model. Predicted results were strongly correlated to measured Hg concentrations (r=0.91,P<0.001), indicating that δ¹⁵N has potential to be used in modeling.     

Kinetics of amino sugar formation from organic residues of different quality

Amino sugars are key compounds of microbial cell walls, which have been widely used as biomarker of microbial residues to investigate soil microbial communities and organic residue cycling processes. However, the formation dynamics of amino sugar is not well understood. In this study, two agricultural Luvisols under distinct tillage managements were amended with uniformly ¹³C-labeled wheat residues of different quality (grain, leaf and root). The isotopic composition of individual amino sugars and CO₂ emission were measured over a 21-day incubation period using liquid chromatography–isotope ratio mass spectrometry (LC–IRMS) and trace gas IRMS. Results showed that, the amount of residue derived amino sugars increased exponentially and reached a maximum within days after residue addition. Glucosamine and galactosamine followed different formation kinetics. The maxima of residue derived amino sugars formation ranged from 14 nmol g⁻¹ dry soil for galactosamine (0.8% of the original concentration) to 319 nmol g⁻¹ dry soil for glucosamine (11% of the original concentration). Mean production times of residue derived amino sugars ranged from 2.1 to 9.3 days for glucosamine and galactosamine, respectively. In general, larger amounts of amino sugars were formed at a higher rate with increasing plant residue quality. The microbial community of the no-till soil was better adapted to assimilate low quality plant residues (i.e. leaf and root). All together, the formation dynamics of microbial cell wall components was component-specific and determined by residue quality and soil microbial community.     

Inter‐specific variability in organic nitrogen uptake of three temperate grassland species

We tested the inter‐specific variability in the ability of three dominant grasses of temperate grasslands to take up organic nitrogen (N) in the form of amino acids in soils of differing fertility. Amino acid uptake was determined by injecting dual labeled glycine‐2‐¹³C‐¹⁵N into the soil, and then measuring the enrichment of both ¹³C and ¹⁵N in plant tissue after 50 hours. We found enrichment of both ¹³C and ¹⁵N in root and shoot material of all species in both soils, providing first evidence for direct uptake of glycine. We show that there was considerable inter‐specific variability in amino acid uptake in the low fertility soil. Here, direct uptake of amino acid was greater in the grass Agrostis capillaris, which typically dominates low fertility grassland, than Lolium perenne, which inhabits more fertile sites. Direct uptake of amino acid for Holcus lanatus. was intermediate between the above two species. Unlike in the low fertility soil, there was no difference in uptake of either ¹³C or ¹⁵N by grasses in the high fertility soil, where uptake of mineral N is thought to be the major mechanism of N uptake of these grasses. Overall, our findings may contribute to our understanding of differences in competitive interactions between grasses in soils of different fertility status.     

青藏高原土壤中15N和全氮形态及其与土壤环境因子间的关系

The patterns of soil nitrogen(N) isotope composition at large spatial and temporal scales and their relationships to environmental factors illustrate N cycle and sources of N,and are integrative indicators of the terrestrial N cycle and its response to global change. The objectives of this study were:i) to investigate the patterns of soil N content and natural abundance of 15N(δ15N) values in different ecosystem types and soil profiles on the Qinghai-Tibetan Plateau; ii) to examine the effects of climatic factors and soil characteristics on the patterns of soil N content and soil δ15N values; and iii) to test the relationship between soil δ15N values and soil C/N ratios across ecosystems and soil profiles. Soil profiles were sampled at 51 sites along two transects 1 875 km in length and 200 km apart and distributed in forest,meadow and steppe on the Qinghai-Tibetan Plateau. Each site was sampled every 10 cm from a soil depth of 0 to 40 cm and each sample was analyzed for soil N content and δ15N values. Our results indicated that soil N and δ15N values(0–40 cm) in meadows were much higher than in desert steppe. Soil N decreased with soil depth for each ecosystem,while variations of soil δ15N values along soil profiles were not statistically significant among most ecosystems but for mountain meadow,lowland meadow,and temperate steppe where soil δ15N values tended to increase with soil depth. The parabolic relationship between soil δ15N values and mean annual precipitation indicated that soil δ15N values increased with increasing precipitation in desert steppe up to 500 mm,and then decreased with increasing precipitation across all other ecosystems. Moreover,the parabolic relationship between δ15N values and mean annual temperature existed in all individual ecosystem types. Soil N and δ15N values(0–40 cm) increased with an increase in soil silt and clay contents. Furthermore,a threshold of C/N ratio of about 11 divided the parabolic relationship between soil δ15N values and soil C/N ratios into positive(C/N 11) and negative(C/N 11) parts,which was valid across all ecosystems and soil profiles. The large explanatory power of soil C/N ratios for soil δ15N values suggested that C and N concentrations,being strongly controlled by precipitation and temperature,were the primary factors determining patterns of soil δ15N on the Qinghai-Tibetan Plateau.     

Isotopic fingerprinting for the authenticity control of crop protection active compounds using the representative insecticide Fipronil

Isotopic fingerprinting was evaluated for its potential to generate characteristic fingerprints of crop protection products in an extensive survey, using the insecticide Fipronil. One hundred and twenty batches of Fipronil from the BASF production site in France were analyzed for the isotope ratios of δ(13)C, δ(15)N, and δ(34)S. Samples spanned a production time of four years and were analyzed by elemental analysis, coupled to isotope ratio mass spectrometry (EA/IRMS). A number of Fipronil samples from other sources were analyzed in the same manner and were compared to the samples from BASF by means of multivariate data analysis. The isotopic fingerprint was sufficiently specific to differentiate between Fipronil from BASF production and Fipronil from other producers. This suggests that isotopic fingerprinting is suitable for the authenticity control of active compounds in crop protection products. It is anticipated that this technique will deliver great benefit in the defense against counterfeits and illegal parallel imports.