Nitrogen and carbon isotope responses of Chinese cabbage and chrysanthemum to the application of liquid pig manure

The effects of the liquid pig manure (LM) used in organic farming on the natural abundance of ¹⁵N and ¹³C signatures in plant tissues have not been studied. We hypothesized that application of LM will (1) increase δ¹⁵N of plant tissues due to the high δ¹⁵N of N in LM as compared with soil N or inorganic fertilizer N, and (2) increase δ¹³C of plant tissues as a result of high salt concentration in LM that decreases stomatal conductance of plants. To test these hypotheses, variations in the δ¹⁵N and δ¹³C of Chinese cabbage (Brassica campestris L.) and chrysanthemum (Chrysanthemum morifolium Ramatuelle) with two different LMs (with δ¹⁵N of +15.6 and +18.2‰) applied at two rates (323 and 646 kg N ha^-1 for cabbage and 150 and 300 kg N ha^-1 for chrysanthemum), or urea (δ¹⁵N = -2.7‰) applied at the lower rate above for the respective species, in addition to the control (no N input) were investigated through a 60-day pot experiment. Application of LM significantly increased plant tissue δ¹⁵N (range +9.4 to +14.9‰) over the urea (+3.2 to +3.3‰) or control (+6.8 to 7.7‰) treatments regardless of plant species, strongly reflecting the δ¹⁵N of the N source. Plant tissue δ¹³C were not affected by the treatments for cabbage (range −30.8 to −30.2‰) or chrysanthemum (−27.3 to −26.8‰). However, cabbage dry matter production decreased while its δ¹³C increased with increasing rate of LM application or increasing soil salinity (P < 0.05), suggesting that salinity stress caused by high rate of LM application likely decreased stomatal conductance and limited growth of cabbage. Our study expanded the use of the δ¹⁵N technique in N source (organic vs. synthetic fertilizer) identification and suggested that plant tissue δ¹³C maybe a sensitive indicator of plant response to salinity stress caused by high LM application rates.     

δ15N of forest soil and understorey vegetation reflect the former agricultural land use

Since the middle of the 19th century, the area covered by forests in France has doubled. These new forests grow on previous agricultural lands. We have studied the influence of this agricultural history on the ¹⁵N abundance of present-day forests planted on farmlands in the Vosges mountains (north-eastern France) between 1898 and 1930. Different types of land use were identified from old cadastres (1814–1836) of 16 farms. Ancient forests adjacent to farmlands were used as controls. Former pastures, meadows, croplands, gardens and ancient forests were compared for soil δ¹⁵N (fraction <50 μm and total soil), C/N, P and N content and fern (Dryopteris carthusiana) δ¹⁵N. The mean δ¹⁵N of soil increased in the order ancient forests (+0.0‰)<pastures (+1.4‰)<croplands (+1.6‰)<meadows (+2.5‰)<gardens (+3.8‰). This increase in soil δ¹⁵N with the intensity of former land use was related to the former input of ¹⁵N-enriched manure, and to an activation of soil nitrification leading to ¹⁵N-depleted nitrate export on previously manured parcels. Fern δ¹⁵N increased in the same order as soil δ¹⁵N in relation to past land use. The mean δ¹⁵N of fern in ancient forests (–4.4‰) and former pastures (–3.4‰) was 5‰ lower than soil δ¹⁵N and the two variables were strongly correlated. The δ¹⁵N of fern in formerly manured parcels varied little (cropland: –2.7‰, meadows: –2.6‰ and gardens: –2.2‰) and independently of soil δ¹⁵N, suggesting that the soil sources of fern N differed between unmanured and manured parcels. Understorey plant δ¹⁵N and soil δ¹⁵N appear to be excellent tracers of previous land use in forests, and could be used in historical studies. The persistence of high isotopic ratios in previously manured parcels, almost a century after afforestation, suggests a long-term influence of former land use on the N cycle in forest soils. Received: 22 January 1999 / Accepted: 22 July 1999     

Effects of land use on 15N natural abundance of soils in Ethiopian highlands

We used the natural abundance of ¹⁵N in soils in forests, pastures and cultivated lands in the Menagesha and Wendo-Genet areas of Ethiopia to make inferences about the N cycles in these ecosystems. Since we have described the history of these sites based on variations in ¹³C natural abundance, patterns of δ¹⁵N and δ¹³C values were compared to determine if shifts of ¹⁵N correlate with shifts of vegetation. At Menagesha, a > 500-yr-old planted forest, we found δ¹⁵N values from −8.8 to +3.5‰ in litter, from −3.5 to +4.5‰ in 0–10 cm soil layer, and from −1.5 to +6.8‰ at >20 cm soil depth. The low δ¹⁵N in litter and surface mineral soils suggests that a closed N cycle has operated for a long time. At this site, the low δ¹³C of the surface horizon and the high δ¹³C of the lower soil horizons is clear evidence of a long phase of C₄ grass dominance or cultivation of C₄ crops before the establishment of the forest >500 years ago. In contrast, at Wendo-Genet, high δ¹³C of soils reveals that most of the land has been uncovered by forests until recently. Soil δ¹⁵N was high throughout (3.4–9.8‰), and there were no major differences between forested, cultivated and pasture soils in δ¹⁵N values of surface mineral soils. The high δ¹⁵N values suggest that open N cycles operate in the Wendo-Genet area. From the points of view of soil fertility management, it is interesting that tall forest ecosystems with relatively closed N cycling could be established on the fairly steep slopes at Menagesha after a long period of grass vegetation cover or cultivation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Isotope ratios and concentrations of sulfur and nitrogen in needles and soils of Picea abies stands as influenced by atmospheric deposition of sulfur and nitrogen compounds

Concentrations and natural isotope abundance of total sulfur and nitrogen as well as sulfate and nitrate concentrations were measured in needles of different age classes and in soil samples of different horizons from a healthy and a declining Norway spruce (Picea abies (L.) Karst.) forest in the Fichtelgebirge (NE Bavaria, Germany), in order to study the fate of atmospheric depositions of sulfur and nitrogen compounds. The mean δ¹⁵N of the needles ranged between −3.7 and −2.1 ‰ and for δ³⁴S a range between −0.4 and +0.9 ‰ was observed. δ³⁴S and sulfur concentrations in the needles of both stands increased continuously with needle age and thus, were closely correlated. The δ¹⁵N values of the needles showed an initial decrease followed by an increase with needle age. The healthy stand showed more negative δ¹⁵N values in old needles than the declining stand. Nitrogen concentrations decreased with needle age. For soil samples at both sites the mean δ¹⁵N and δ³⁴S values increased from −3 ‰ (δ¹⁵N) or +0.9 ‰ (δ³⁴S) in the uppermost organic layer to about +4 ‰ (δ¹⁵N) or +4.5 ‰ (δ³⁴S) in the mineral soil. This depth-dependent increase in abundance of ¹⁵N and ³⁴S was accompanied by a decrease in total nitrogen and sulfur concentrations in the soil. δ¹⁵N values and nitrogen concentrations were closely correlated (slope −0.0061 ‰ δ¹⁵N per μmol eq N g_dw ⁻¹), and δ³⁴S values were linearly correlated with sulfur concentrations (slope −0.0576 ‰ δ³⁴S per μmol eq S g_dw ⁻¹). It follows that in the same soil samples sulfur concentrations were linearly correlated with the nitrogen concentrations (slope 0.0527), and δ³⁴S values were linearly correlated with δ¹⁵N values (slope 0.459). A correlation of the sulfur and nitrogen isotope abundances on a Δ basis (which considers the different relative frequencies of ¹⁵N and ³⁴S), however, revealed an isotope fractionation that was higher by a factor of 5 for sulfur than for nitrogen (slope 5.292). These correlations indicate a long term synchronous mineralization of organic nitrogen and sulfur compounds in the soil accompanied by element-specific isotope fractionations. Based on different sulfur isotope abundance of the soil (δ³⁴S=0.9 ‰ for total sulfur of the organic layer was assumed to be equivalent to about −1.0 ‰ for soil sulfate) and of the atmospheric SO₂ deposition (δ³⁴S=2.0 ‰ at the healthy site and 2.3 ‰ at the declining site) the contribution of atmospheric SO₂ to total sulfur of the needles was estimated. This contribution increased from about 20 % in current-year needles to more than 50 % in 3-year-old needles. The proportion of sulfur from atmospheric deposition was equivalent to the age dependent sulfate accumulation in the needles. In contrast to the accumulation of atmospheric sulfur compounds nitrogen compounds from atmospheric deposition were metabolized and were used for growth. The implications of both responses to atmospheric deposition are discussed.     

Isotopic signature of nitrate in two contrasting watersheds of Brush Brook,Vermont, USA

We used the dual isotope method to study differences in nitrate export in two subwatersheds in Vermont, USA. Precipitation, soil water and streamwater samples were collected from two watersheds in Camels Hump State Forest, located within the Green Mountains of Vermont. These samples were analyzed for the δ¹⁵N and δ¹⁸O of NO₃⁻. The range of δ¹⁵N–NO₃⁻ values overlapped, with precipitation −4.5‰ to +2.0‰ (n = 14), soil solution −10.3‰ to +6.2‰ (n = 12) and streamwater +0.3‰ to +3.1‰ (n = 69). The δ¹⁸O of precipitation NO₃⁻ (mean 46.8 ± 11.5‰) was significantly different (P < 0.001) from that of the stream (mean 13.2 ± 4.3‰) and soil waters (mean 14.5 ± 4.2‰) even during snowmelt periods. Extracted soil solution and streamwater δ¹⁸O of NO₃⁻ were similar and within the established range of microbially produced NO₃⁻, demonstrating that NO₃⁻ was formed by microbial processes. The δ¹⁵N and δ¹⁸O of NO₃⁻ suggests that although the two tributaries have different seasonal NO₃⁻ concentrations, they have a similar NO₃⁻ source.     

Effect of C3–C4 Vegetation Change on δ13C and δ15N Values of Soil Organic Matter Fractions Separated by Thermal Stability

Carbon isotopic composition of soils subjected to C₃–C₄ vegetation change can be used to estimate C turnover in bulk soil and in soil organic matter (SOM) pools with fast and intermediate turnover rates. We hypothesized that the biological availability of SOM pools is inversely proportional to their thermal stability, so that thermogravimetry can be used to separate SOM pools with contrasting turnover rates. Soil samples from a field plot cultivated for 10.5 years with the perennial C₄ plant Miscanthus×gigantheus were analyzed by thermogravimetry coupled with differential scanning calorimetry (DSC). Three SOM fractions were distinguished according to the differential weight losses and exothermic or endothermic reactions measured by DSC. The δ¹³C and δ¹⁵N values of these three fractions obtained by gradual soil heating were measured by IRMS. The weight losses up to 190 °C mainly reflected water evaporation because no significant C and N losses were detected and δ¹³C and δ¹⁵N values of the residual SOM remained unchanged. The δ¹³C values (−16.4‰) of SOM fraction decomposed between 190 and 390 °C (containing 79% of total soil C) were slightly closer to that of the Miscanthus plant tissues (δ¹³C = −11.8‰) compared to the δ¹³C values (−16.8‰) of SOM fraction decomposed above 390 °C containing the residual 21% of SOM. Thus, the C turnover in the thermally labile fraction was faster than that in thermally stable fractions, but the differences were not very strong. Therefore, in this first study combining TG-DSC with isotopic analysis, we conclude that the thermal stability of SOM was not very strongly related to biological availability of SOM fractions. In contrast to δ¹³C, the δ¹⁵N values strongly differed between SOM fractions, suggesting that N turnover in the soil was different from C turnover. More detailed fractionation of SOM by thermal analysis with subsequent isotopic analysis may improve the resolution for δ¹³C.     

Natural 15N abundances of maize and soil amended with urea and composted pig manure

To investigate the effect of inorganic fertilizer and composted manure amendments on the N isotope composition (delta ¹⁵N) of crop and soil, maize (Zea mays L.) was cultivated under greenhouse conditions for 30, 40, 50, 60, and 70 days. Composted pig manure (delta ¹⁵N= +13.9) and urea (-2.3) were applied at 0 and 0 kg N ha^–1 (C0U0), 0 and 150 kg N ha^–1 (C0U2), 150 and 0 kg N ha^–1 (C2U0), and 75 and 75 kg N ha^–1 (C1U1), respectively. The delta ¹⁵N of total soil-N was not affected by both amendments, but delta ¹⁵N of NH⁺ ₄ and NO^– ₃ provided some information on the N isotope fractionation in soil. During the early growth stage, significant differences (P < 0.05) in delta ¹⁵N among maize subjected to different treatments were observed. After 30 days of growth, the delta ¹⁵N values of maize were +6.6 for C0U0, +1.1 for C0U2, +7.7 for C2U0, and +4.5 for C1U1. However, effects of urea and composted manure application on maize delta ¹⁵N progressively decreased with increasing growth period, probably due to isotope fractionation accompanying N losses and increased uptake of soil-derived N by maize. After 70 days of growth, delta ¹⁵N of leaves and grains of maize amended with composted pig manure were significantly (P < 0.05) higher than those with urea. The temporal variations in delta ¹⁵N of maize amended with urea and composted manure indicate that plant delta ¹⁵N is generally not a good tracer for N sources applied to field. Our data can be used in validation of delta ¹⁵N fractionation models in relation to N source inputs.     

Short-range spatial variability of soil δ15N natural abundance – effects on symbiotic N2-fixation estimates in pea

The δ¹⁵N natural abundance (‰) of the total soil N pool varies at the landscape level, but knowledge on short-range variability and consequences for the reliability of isotopic methods are poorly understood. The short-range spatial variability of soil δ¹⁵N natural abundance as revealed by the ¹⁵N abundance in spring barley and N₂-fixing pea was measured within the 0.15–4 m scale at flowering and at maturity. The short-range spatial variability of soil δ¹⁵N natural abundance and symbiotic nitrogen fixation were high at both growth stages. Along a 4-m row, the δ¹⁵N natural abundance in barley reference plants varied up to 3.9‰, and sometimes this variability was observed even between plants grown only 30 cm apart. The δ¹⁵N natural abundance in pea varied up to 1.4‰ within the 4-m row. The estimated percentage of nitrogen derived from the atmosphere (%Ndfa) varied from 73–89% at flowering and from 57–95% at maturity. When increasing the sampling area from 0.01 m² (single plants) and up to 0.6 m² (14 plants) the %Ndfa coefficient of variation (CV) declined from 5 to 2% at flowering and from 12 to 2% at maturity. The implications of the short-range variability in δ¹⁵N natural-abundance are that estimates of symbiotic N₂-fixation can be obtained from the natural abundance method if at least half a square meter of crop and reference plants is sampled for the isotopic analysis. In fields with small amounts of representative reference crops (weeds) it might be necessary to sow in reference crop species to secure satisfying N₂-fixation estimates.     

Monitoring plant physiological characteristics to evaluate mine site revegetation: A case study from the wet-dry tropics of northern Australia

Biologically driven markers or monitors were used to evaluate plant and ecosystem health of uranium-mining affected sites. Plant water, nitrogen (N) and phosphorus (P) status were used to measure physiological characteristics of tree and shrub species at sites perturbed by mining activities (waste rock dumps: WRD 1, WRD 2; mine wastewater irrigated woodland) and of species at undisturbed woodland (tropical savanna). Plant water status was evaluated by measuring leaf relative water content (RWC) and carbon isotope discrimination (δ¹³C). Leaf RWC varied significantly (P<0.0001) between wet and dry season in species at the woodland sites with higher RWC in the wet season compared to the dry season. No seasonal differences were observed in RWC in species at the WRDs. Leaf δ¹³C was similar in species at woodland sites and WRD 2 (−28.8 to −28.1‰) but was significantly (P<0.05) lower in species at WRD 1 (−27.6‰). This suggests that species at WRD 1 had a lower water availability and/or lower water use compared to species at all other sites. WRD substrate had an up to 4-orders of magnitude greater availability of inorganic phosphate (Pi) compared to woodland soil as determined using in situ ion exchange resin. Pi concentrations in xylem sap of species at WRDs were 2- to 3-fold higher compared to species at woodland sites. Plant nitrate reductase (NR) activity was low in most species at woodland and WRD 1. In contrast, Eucalyptus and Acacia species had high NR activities of up to 300–700 pkat g^-1 fw at WRD 2 indicating that these species had greater nitrate use than species at all other sites. Nitrate availability in the top five cm of the profile, as determined using in situ ion exchange resins, increased at all sites in the wet season, but no significant differences were observed between sites using this method. However, traditional soil analysis revealed that WRD substrate had a 2-times higher nitrate content (0 to 1000 mm depth) compared to woodland soil. Thus, it is likely that plants at WRD2 accessed nitrate from deeper parts of the profile. Proline, an indicator of plant stress, was found in appreciable quantities in leaves of herbaceous species but not in woody species. Soil and leaf δ¹⁵N were measured to investigate N-cycling and the contribution of diazotrophic N₂ fixation to plant N nutrition. Soil δ¹⁵N values were highest and most variable at WRD 2 (6.2‰) compared to all other sites (irrigated woodland 3.1‰, undisturbed woodland 2.5‰, WRD 1 0.9‰). This may indicate that N-turnover and nitrification was greatest at WRD 2 leading to greater ¹⁵N enrichment of soil N. At all sites, Acacia species were nodulated and putatively fixing N₂. With the exception of WRD 2 where leaf δ¹⁵N of Acacia species averaged 0.9‰, Acacia species had ¹⁵N depleted values characteristic of species that receive N derived from N₂ fixation (−0.8 to −0.6‰). Eucalyptus species at the woodland also had ¹⁵N depleted values (average −0.4‰) but ¹⁵N enriched values (0.3 to 1.8‰) at the three mining affected sites. The results show that for the plants studied foliar δ¹⁵N could not be used as an unequivocal measure of plant N sources. The results suggest that biomonitoring of plant and ecosystem health has potential in evaluating performance of mine site revegetation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Natural 15N abundance of soil N pools and N2O reflect the nitrogen dynamics of forest soils

Natural ¹⁵N abundance measurements of ecosystem nitrogen (N) pools and ¹⁵N pool dilution assays of gross N transformation rates were applied to investigate the potential of δ¹⁵N signatures of soil N pools to reflect the dynamics in the forest soil N cycle. Intact soil cores were collected from pure spruce (Picea abies (L.) Karst.) and mixed spruce-beech (Fagus sylvatica L.) stands on stagnic gleysol in Austria. Soil δ¹⁵N values of both forest sites increased with depth to 50 cm, but then decreased below this zone. δ¹⁵N values of microbial biomass (mixed stand: 4.7 ± 0.8‰, spruce stand: 5.9 ± 0.9‰) and of dissolved organic N (DON; mixed stand: 5.3 ± 1.7‰, spruce stand: 2.6 ± 3.3‰) were not significantly different; these pools were most enriched in ¹⁵N of all soil N pools. Denitrification represented the main N₂O-producing process in the mixed forest stand as we detected a significant ¹⁵N enrichment of its substrate NO₃⁻ (3.6 ± 4.5‰) compared to NH₄⁺ (−4.6 ± 2.6‰) and its product N₂O (−11.8 ± 3.2‰). In a ¹⁵N-labelling experiment in the spruce stand, nitrification contributed more to N₂O production than denitrification. Moreover, in natural abundance measurements the NH₄⁺ pool was slightly ¹⁵N-enriched (−0.4 ± 2.0 ‰) compared to NO₃⁻ (−3.0 ± 0.6 ‰) and N₂O (−2.1 ± 1.1 ‰) in the spruce stand, indicating nitrification and denitrification operated in parallel to produce N₂O. The more positive δ¹⁵N values of N₂O in the spruce stand than in the mixed stand point to extensive microbial N₂O reduction in the spruce stand. Combining natural ¹⁵N abundance and ¹⁵N tracer experiments provided a more complete picture of soil N dynamics than possible with either measurement done separately.     

Disentangling effects of growth and nutritional status on seabird stable isotope ratios

A growing number of studies suggest that an individual’s physiology affects its carbon and nitrogen stable isotope signatures, obscuring a signal often assumed to be only a reflection of diet and foraging location. We examined effects of growth and moderate food restriction on red blood cell (RBC) and feather δ¹⁵N and δ¹³C in rhinoceros auklet chicks (Cerorhinca monocerata), a piscivorous seabird. Chicks were reared in captivity and fed either control (75 g/day; n = 7) or ~40% restricted (40 g/day; n = 6) amounts of high quality forage fish. We quantified effects of growth on isotopic fractionation by comparing δ¹⁵N and δ¹³C in control chicks to those of captive, non-growing subadult auklets (n = 11) fed the same diet. To estimate natural levels of isotopic variation, we also collected blood from a random sample of free-living rhinoceros auklet adults and chicks in the Gulf of Alaska (n = 15 for each), as well as adult feather samples (n = 13). In the captive experiment, moderate food restriction caused significant depletion in δ¹⁵N of both RBCs and feathers in treatment chicks compared to control chicks. Growth also induced depletion in RBC δ¹⁵N, with chicks exhibiting lower δ¹⁵N when they were growing the fastest. As growth slowed, δ¹⁵N increased, resulting in an overall pattern of enrichment over the course of the nestling period. Combined effects of growth and restriction depleted δ¹⁵N in chick RBCs by 0.92‰. We propose that increased nitrogen-use efficiency is responsible for ¹⁵N depletion in both growing and food-restricted chicks. δ¹⁵N values in RBCs of free-ranging auklets fell within a range of only 1.03‰, while feather δ¹⁵N varied widely. Together, our captive and field results suggest that both growth and moderate food restriction can affect stable isotope ratios in an ecologically meaningful way in RBCs although not feathers due to greater natural variability in this tissue.     

The intramolecular δ<Superscript>15</Superscript>N of lysine responds to respiratory status in <Emphasis Type="Italic">Paracoccus denitrificans</Emphasis>

Summary. Presented here is the first experimental evidence that natural, intramolecular, isotope ratios are sensitive to physiological status, based on observations of intramolecular δ¹⁵N of lysine in the mitochondrial mimic Paracoccus denitrificans. Paracoccus denitrificans, a versatile, gram-negative bacterium, was grown either aerobically or anaerobically on isotopically-characterized ammonium as sole cell-nitrogen source. Nitrogen isotope composition of the biomass with respect to source ammonium was = −6.2 ± 1.2‰ for whole cells under aerobic respiration, whereas cells grown anaerobically produced no net fractionation ( = −0.3 ± 0.23‰). Fractionation of ¹⁵N between protein nitrogen and total cell nitrogen increased during anaerobic respiration and suggests that residual nitrogen-containing compounds in bacterial cell membranes are isotopically lighter under anaerobic respiration. In aerobic cells, the lysine intramolecular difference between peptide and sidechain nitrogen is negligible, but in anaerobic cells was a remarkable Δ¹⁵N_{p − s} = δ¹⁵N_peptide − δ¹⁵N_sidechain = +11.0‰, driven predominantly by enrichment at the peptide N. Consideration of known lysine pathways suggests this to be likely due to enhanced synthesis of peptidoglycans in the anaerobic state. These data indicate that distinct pathway branching ratios associated with microbial respiration can be detected by natural intramolecular Δδ¹⁵N measurements, and are the first in vivo observations of position-specific measurements of nitrogen isotope fractionation.     

Boron uptake in sunflower,squash and cultured tobacco cells: Studies with stable isotope and ICP-MS

A direct correlation was found between fractional losses of added N and the change in δ¹⁵N‰ during 19 years in an experiment with annual additions of N at three rates to a Scots pine (Pinus sylvestris L.) forest in northern Sweden. This confirms that processes leading to losses of N discriminate against¹⁵N, and opens possibilities to conduct retrospective studies of the N balance in forests.     

Estimation of symbiotically fixed nitrogen in field grown soybeans: An application of natural15N/14N abundance and a low level15N-tracer technique

Summary Plants from agricultural and natural upland ecosystem were investigated for¹⁵N content to evaluate the role of symbiotic N₂-fixation in the nitrogen nutrition of soybean. Increased yields and lower δ¹⁵N values of nodulating soybeansvs, non-nodulating isolines gave semi-quantitative estimates of N₂ fixation. A fairly large discrepancy was found between estimations by δ¹⁵N and by N yield at 0 kg N/ha of fertilizer. More precise estimates were made by following changes in plant δ¹⁵N when fertilizer δ¹⁵N was varied near¹⁵N natural abundance level. Clearcut linear relationships between δ¹⁵N values of whole plants and of fertilizer were obtained at 30 kg N/ha of fertilizer for three kinds of soils. In experimental field plots, nodulating soybeans obtained 13±1% of their nitrogen from fertilizer, 66±8% from N₂ fixation and 21±10% from soil nitrogen in Andosol brown soil; 30%, 16% and 54% in Andosol black soil; 7%, 77% and 16% in Alluvial soil, respectively. These values for N₂ fixation coincided with each corresponding estimation by N yield method. Other results include: 1)¹⁵N content in upland soils and plants was variable, and may reflect differences in the mode of mineralization of soil organics, and 2) nitrogen isotopic discrimination during fertilizer uptake (δ¹⁵N of plant minus fertilizer) ranged from −2.2 to +4.9‰ at 0–30 kg N/ha of fertilizer, depending on soil type and plant species. The proposed method can accurately and relatively simply establish the importance of symbiotic nitrogen fixation for soybeans growing in agricultural settings.     

Immobilisation, remineralisation and residual effects in subsequent crops of dairy cattle slurry nitrogen compared to mineral fertiliser nitrogen

About 50–60% of dairy cattle slurry nitrogen is ammonium N. Part of the ammonium N in cattle slurry is immobilised due to microbial decomposition of organic matter in the slurry after application to soil. The immobilisation and the remineralisation influence the fertiliser value of slurry N and the amount of organic N that is retained in soil. The immobilisation and the remineralisation of 15 N-labelled dairy cattle slurry NH₄-N were studied through three growing seasons after spring application under temperate conditions. Effects of slurry distribution (mixing, layer incorporation, injection, surface-banding) and extra litter straw in the slurry on the plant utilisation of labelled NH₄-N from slurry were studied and compared to the utilisation of ¹⁵N-labelled mineral fertiliser. The initial immobilisation of slurry N was influenced by the slurry distribution in soil. More N was immobilised when the slurry was mixed with soil. Surface-banding of slurry resulted in significant volatilisation losses and less residual ¹⁵N in soil. Much more N was immobilised after slurry incorporation than after mineral fertiliser application. After 2.5 years the recovery of labelled N in soil (0–25 cm) was 46% for slurry mixed with soil, 42% for injected slurry, 22% for surface-banded slurry and 24% for mineral fertiliser N. The total N uptake in a ryegrass cover crop was 5–10 kg N/ha higher in the autumn after spring-application of cattle slurry (100–120 kg NH₄-N/ha) compared to the mineral fertiliser N reference, but the immobilised slurry N (labelled N) only contributed little to the extra N uptake in the autumn. Even in the second autumn after slurry application there was an extra N uptake in the cover crop (0–10 kg N/ha). The residual effect of the cattle slurry on spring barley N uptake was insignificant in the year after slurry application (equivalent to 3% of total slurry N). Eighteen months after application, 13% of the residual ¹⁵N in soil was found in microbial biomass whether it derived from slurry or mineral fertiliser, but the remineralisation rate (% crop removal of residual ¹⁵N) was higher for fertiliser- than for slurry-derived N, except after surface-banding. Extra litter straw in the slurry had a negligible influence on the residual N effects in the year after application. It is concluded that a significant part of the organic N retained in soil after cattle slurry application is derived from immobilised ammonium N, but already a few months after application immobilised N is stabilised and only slowly released. The immobilised N has negligible influence on the residual N effect of cattle slurry in the first years after slurry application, and mainly contributes to the long-term accumulation of organic N in soil together with part of the organic slurry N. Under humid temperate conditions the residual N effects of the manure can only be optimally utilised when soil is also covered by plants in the autumn, because a significant part of the residual N is released in the autumn, and there is a higher risk of N leaching losses on soils that receive cattle slurry regularly compared to soils receiving only mineral N fertilisers.     

Nitrogen-15 balances for urea and neem coated urea applied to lowland rice following two cowpea cropping systems

Little is known about whether the high N losses from inorganic N fertilizers applied to lowland rice (Oryza sativa L.) are affected by the combined use of either legume green manure or residue with N fertilizers. Field experiments were conducted in 1986 and 1987 on an Andaqueptic Haplaquoll in the Philippines to determine the effect of cowpea [Vigna unguiculata (L.) Walp.] cropping systems before rice on the fate and use efficiency of¹⁵N-labeled, urea and neem cake (Azadirachta indica Juss.) coated urea (NCU) applied to the subsequent transplanted lowland rice crop. The pre-rice cropping systems were fallow, cowpea incorporated at the flowering stage as a green manure, and cowpea grown to maturity with subsequent incorporation of residue remaining after grain and pod removal. The incorporated green manure contained 70 and 67 kg N ha⁻¹ in 1986 and 1987, respectively. The incorporated residue contained 54 and 49 kg N ha⁻¹ in 1986 and 1987, respectively. The unrecovered¹⁵N in the¹⁵N balances for 58 kg N ha⁻¹ applied as urea or NCU ranged from 23 to 34% but was not affected by pre-rice cropping system. The partial pressure of ammoniapNH₃, and floodwater (nitrate + nitrite)-N following application of 29 kg N ha⁻¹ as urea or NCU to 0.05-m-deep floodwater at 14 days after transplanting was not affected by pre-rice cropping system. In plots not fertilized with urea or NCU, green manure contributed an extra 12 and 26 kg N ha⁻¹, to mature rice plants in 1986 and 1987, respectively. The corresponding contributions from residue were 19 and 23 kg N ha⁻¹, respectively. Coating urea with 0.2g neem cake per g urea had no effect on loss of urea-N in either year; however, it significantly increased grain yield (0.4 Mg ha⁻¹) and total plant N (11 kg ha⁻¹) in 1987 but not in 1986.     

Stable Isotope Ratios as a Tool for Assessing Changes in Carbon and Nutrient Sources in Antarctic Terrestrial Ecosystems

Samples of an angiosperm species, nine lichen species and a terrestrial alga, were collected from a variety of Antarctic terrestrial habitats, and were analysed for C and N stable isotope composition. Collections were made along natural gradients, the marine gradient, running from the sea coast inland and the moisture gradient, determined by melt water and precipitation runoff, and running towards the sea coast. Considerable variation in stable isotope ratios was found; δ¹³C values ranged between −16 and −32‰ and δ¹⁵N values between −23 and +23‰ The variation in stable carbon isotope ratios could be attributed in part to species specific differences, but differences in water availability also played a role, as was shown for the terrestrial alga Prasiola crispa and the lichen species Usnea antarctica. The differences in the isotope ratios of nitrogen could be retraced to the origin of nitrogen: marine or terrestrial. The nitrogen stable isotope ratios were influenced by both the marine gradient from the sea inland and the melt water and precipitation flow running in the opposite direction, towards the sea. This was shown for the lichen species Turgidosculum complicatulum and the angiosperm species Deschampsia antarctica. The variation in the C and N stable isotope ratios can be used to determine sources and pathways of N and changes in the water availability in Antarctic terrestrial ecosystems. Contrary to earlier reports the use of stable N isotope ratios is possible in this case because of the relative simplicity of the structure of the Antarctic terrestrial ecosystems.     

Variations and controls of nitrogen stable isotopes in particulate organic matter of lakes

Nitrogen stable isotope (δ¹⁵N) data of particulate organic matter (POM) from the literature were analyzed to provide an understanding of the variations and controls of δ¹⁵N_POM in lakes at the global scale. The δ¹⁵N_POM variability characterized by seasonal mean, minimum, maximum, and amplitude (defined as δ¹⁵N_POM maximum − δ¹⁵N_POM minimum) from 36 lakes with seasonal data did not change systematically with latitude, but was significantly lower in small lakes than in large lakes. The seasonal mean δ¹⁵N_POM increased from oligotrophic lakes to eutrophic lakes despite large variations that are attributed to the occurrences of nitrogen fixation across the trophic gradient and the differences in δ¹⁵N of dissolved inorganic nitrogen (DIN) in individual lakes. Seasonal mean δ¹⁵N_POM was significantly correlated with DIN concentration and δ¹⁵N_DIN in two subsets of lakes. Seasonal minimum δ¹⁵N_POM in individual lakes is influenced by nitrogen fixation and δ¹⁵N_DIN while seasonal maximum δ¹⁵N_POM is influenced by lake trophic state and δ¹⁵N_DIN. As a result of the dominance of non-living POM in the unproductive surface waters, seasonal δ¹⁵N_POM amplitude was small (mean = 4.2‰) in oligotrophic lakes of all latitudes. On the other hand, seasonal δ¹⁵N_POM amplitude in eutrophic lakes was large (mean = 10.3‰), and increased from low to high latitudes, suggesting that the seasonal variability of δ¹⁵N in the phytoplankton-dominated POM pool was elevated by the greater spans of solar radiation and thermal regimes at high latitudes. The δ¹⁵N_POM from 42 lakes with no seasonal data revealed no consistent patterns along latitude, lake area, and trophic gradients, and a greater than 2‰ depletion compared to the lakes with seasonal data. Along with the large seasonal variability of δ¹⁵N_POM within lakes, these results provide insightful information on sampling design for the studies of food web baseline in lakes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Natural abundance of 15N in tropical plants with emphasis on tree legumes

Natural abundance of ¹⁵N ( ¹⁵N) of leaves harvested from tropical plants in Brazil and Thailand was analyzed. The ¹⁵N values of non-N₂-fixing trees in Brazil were +4.5±1.9, which is lower than those of soil nitrogen (+8.0±2.2). In contrast, mimosa and kudzu had very low ¹⁵N values (–1.4+0.5). The ¹⁵N values of Panicum maximum and leguminous trees, except Leucaena leucocephala, were similar to those of non-N₂-fixing trees, suggesting that the contribution of fixed N in these plants is negligible. The ¹⁵N values of non-N₂-fixing trees in Thailand were +4.9±2.0. Leucaena leucocephala, Sesbania grandiflora, Casuarina spp. and Cycas spp. had low ¹⁵N values, close to the value of atmospheric N₂ (0), pointing to a major contribution of N₂ fixation in these plants. Cassia spp. and Tamarindus indica had high ¹⁵N values, which confirms that these species are non-nodulating legumes. The ¹⁵N values of Acacia spp. and Gliricidia sepium and other potentially nodulating tree legumes were, on average, slightly lower than those of non-N₂-fixing trees, indicating a small contribution of N₂ fixation in these legumes.     

Turnover rates of nitrogen stable isotopes in the salt marsh mummichog, Fundulus heteroclitus, following a laboratory diet switch

Nitrogen stable isotopes are frequently used in ecological studies to estimate trophic position and determine movement patterns. Knowledge of tissue-specific turnover and nitrogen discrimination for the study organisms is important for accurate interpretation of isotopic data. We measured δ¹⁵ N turnover in liver and muscle tissue in juvenile mummichogs, Fundulus heteroclitus, following a laboratory diet switch. Liver tissue turned over significantly faster than muscle tissue suggesting the potential for a multiple tissue stable isotope approach to study movement and trophic position over different time scales; metabolism contributed significantly to isotopic turnover for both liver and muscle. Nitrogen diet-tissue discrimination was estimated at between 0.0 and 1.2‰ for liver and –1.0 and 0.2‰ for muscle. This is the first experiment to demonstrate a significant variation in δ¹⁵ N turnover between liver and muscle tissues in a fish species.