Natural Abundance of <Superscript>15</Superscript>N in Nitrogen-Limited Forests and Tundra Can Estimate Nitrogen Cycling Through Mycorrhizal Fungi: A Review

The hyphae of ectomycorrhizal and ericoid mycorrhizal fungi proliferate in nitrogen (N)-limited forests and tundra where the availability of inorganic N is low; under these conditions the most common fungal species are those capable of protein degradation that can supply their host plants with organic N. Although it is widely understood that these symbiotic fungi supply N to their host plants, the transfer is difficult to quantify in the field. A novel approach uses the natural ¹⁵N:¹⁴N ratios (expressed as δ¹⁵N values) in plants, soils, and mycorrhizal fungi to estimate the fraction of N in symbiotic trees and shrubs that enters through mycorrhizal fungi. This calculation is possible because mycorrhizal fungi discriminate against ¹⁵N when they create compounds for transfer to plants; host plants are depleted in ¹⁵N, whereas mycorrhizal fungi are enriched in ¹⁵N. The amount of carbon (C) supplied to these fungi can be stoichiometrically calculated from the fraction of plant N derived from the symbiosis, the N demand of the plants, the fungal C:N ratio, and the fraction of N retained in the fungi. Up to a third of C allocated belowground, or 20% of net primary production, is used to support ectomycorrhizal fungi. As anthropogenic N inputs increase, the C allocation to fungi decreases and plant δ¹⁵N increases. Careful analyses of δ¹⁵N patterns in systems dominated by ectomycorrhizal and ericoid mycorrhizal symbioses may reveal the ecosystem-scale effects of alterations in the plant–mycorrhizal symbioses caused by shifts in climate and N deposition. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Foliar <Superscript>15</Superscript>N natural abundance indicates phosphorus limitation of bog species

Foliar δ¹⁵N, %N and %P in the dominant woody and herbaceous species across nutrient gradients in New Zealand restiad (family Restionaceae) raised bogs revealed marked differences in plant δ¹⁵N correlations with P. The two heath shrubs, Leptospermum scoparium (Myrtaceae) and Dracophyllum scoparium (Epacridaceae), showed considerable isotopic variation (−2.03 to −15.55‰, and −0.39 to −12.06‰, respectively) across the bogs, with foliar δ¹⁵N strongly and positively correlated with P concentrations in foliage and peat, and negatively correlated with foliar N:P ratios. For L. scoparium, the isotopic gradient was not linked to ectomycorrhizal (ECM) fractionation as ECMs occurred only on higher nutrient marginal peats where ¹⁵N depletion was least. In strong contrast, restiad species (Empodisma minus Sporadanthus ferrugineus, S. traversii) showed little isotopic variation across the same nutrient gradients. Empodisma minus and S. traversii had δ¹⁵N levels consistently around 0‰ (means of −0.12‰ and +0.15‰ respectively), and S. ferrugineus, which co-habited with E. minus, was more depleted (mean −4.97‰). The isotopic differences between heath shrubs and restiads were similar in floristically dissimilar bogs and may be linked to contrasting nutrient demands, acquisition mechanisms, and root morphology. Leptospermum scoparium shrubs on low nutrient peats were stunted, with low tissue P concentrations, and high N:P ratios, suggesting they were P-limited, which was probably exacerbated by markedly reduced mycorrhizal colonisations. The coupling of δ¹⁵N depletion and %P in heath shrubs suggests that N fractionation is promoted by P limitation. In contrast, the constancy in δ¹⁵N of the restiad species through the N and P gradients suggests that these are not suffering from P limitation.     

Isotopic signatures and trophic status of Ramaria

The genus Ramaria is composed of several subgenera that often correspond to specific trophic strategies. Because carbon and nitrogen isotopes can be used to assess fungal trophic status and nitrogen sources, we accordingly carried out an extensive survey of isotopic patterns in archived specimens of Ramaria from Germany and other locations. Isotopic patterns in species generally corresponded to subgeneric affiliations and to the range of different potential substrates, with fungi fruiting on wood and litter (subgenera Asteroramaria and Lentoramaria) much lower in δ¹⁵N (≈−3‰) than ectomycorrhizal taxa (≈12‰) (subgenus Ramaria) or taxa fruiting on soil (≈13‰) (subgenus Echinoramaria). Conversely, fungi fruiting on wood and litter were higher in δ¹³C (−23‰) than those fruiting on soil (≈−27‰), with ectomycorrhizal fungi intermediate (≈−24.5‰). Fungi colonizing mineral soil horizons were about 3‰ enriched in ¹⁵N relative to those colonizing both mineral and organic horizons. The high δ¹⁵N and low δ¹³C signatures of taxa fruiting on soil remains unexplained. The high degree of fidelity of isotopic signatures with subgeneric classifications and life history traits suggests that sporocarps are good integrators of patterns of carbon and nitrogen cycling for specific taxa. Archived specimens represent a useful trove of life history information that could be mined without requiring extensive supporting isotopic data from other ecosystem pools.     

Correlations between foliar δ15N and nitrogen concentrations may indicate plant-mycorrhizal interactions

Nitrogen isotope measurements may provide insights into changing interactions among plants, mycorrhizal fungi, and soil processes across environmental gradients. Here, we report changes in δ¹⁵N signatures due to shifts in species composition and nitrogen (N) dynamics. These changes were assessed by measuring fine root biomass, net N mineralization, and N concentrations and δ¹⁵N of foliage, fine roots, soil, and mineral N across six sites representing different post-deglaciation ages at Glacier Bay, Alaska. Foliar δ¹⁵N varied widely, between 0 and –2‰ for nitrogen-fixing species, between 0 and –7‰ for deciduous non-fixing species, and between 0 and –11‰ for coniferous species. Relatively constant δ¹⁵N values for ammonium and generally low levels of soil nitrate suggested that differences in ammonium or nitrate use were not important influences on plant δ¹⁵N differences among species at individual sites. In fact, the largest variation among plant δ¹⁵N values were observed at the youngest and oldest sites, where soil nitrate concentrations were low. Low mineral N concentrations and low N mineralization at these sites indicated low N availability. The most plausible mechanism to explain low δ¹⁵N values in plant foliage was a large isotopic fractionation during transfer of nitrogen from mycorrhizal fungi to plants. Except for N-fixing plants, the foliar δ¹⁵N signatures of individual species were generally lower at sites of low N availability, suggesting either an increased fraction of N obtained from mycorrhizal uptake (f), or a reduced proportion of mycorrhizal N transferred to vegetation (T _r). Foliar and fine root nitrogen concentrations were also lower at these sites. Foliar N concentrations were significantly correlated with δ¹⁵N in foliage of Populus, Salix, Picea, and Tsuga heterophylla, and also in fine roots. The correlation between δ¹⁵N and N concentration may reflect strong underlying relationships among N availability, the relative allocation of carbon to mycorrhizal fungi, and shifts in either f or T _r. Received: 14 December 1998 / Accepted: 16 August 1999     

Isotopic signature (<Superscript>15</Superscript>N/<Superscript>14</Superscript>N and <Superscript>13</Superscript>C/<Superscript>12</Superscript>C) confirms similarity of trophic niches of millipedes (Myriapoda,diplopoda) in a temperate deciduous forest

The species composition, abundance, and isotopic signature of millipedes (Myriapoda, Diplopoda) were investigated in seven biotopes of Kaluzhskie Zaseki State Nature Reserve. Nine Diplopoda species were found in total, and the local species diversity (within a sampling plot) reached seven species. The Diplopoda tissues were similar to the plant litter in the isotopic composition of nitrogen (δ¹⁵N was by 0.4‰ higher, on average), but were strongly enriched in heavy carbon (δ¹³C was by 4‰ higher, on average). Removal of mineral carbon from the cuticle reduced δ¹³C of Diplopoda by about 1.4‰ on average. Differences in the δ¹⁵N and δ¹³C values between the species did not exceed 2.5‰. Differences in the isotopic compositions of the considered species were small, and, it is impossible to distinguish particular trophic guilds in the Diplopoda community. Analysis of the published data confirmed that isotopic differentiation of millipedes was much less pronounced than in other investigated groups of soil animals. Hence, millipedes of the deciduous forest form a uniform trophic group.     

Interpretation of nitrogen isotope signatures using the NIFTE model

Nitrogen cycling in forest soils has been intensively studied for many years because nitrogen is often the limiting nutrient for forest growth. Complex interactions between soil, microbes, and plants and the consequent inability to correlate δ¹⁵N changes with biologic processes have limited the use of natural abundances of nitrogen isotopes to study nitrogen (N) dynamics. During an investigation of N dynamics along the 250-year-old successional sequence in Glacier Bay, Alaska, United States, we observed several puzzling isotopic patterns, including a consistent decline in δ¹⁵N of the late successional dominant Picea at older sites, a lack of agreement between mineral N δ¹⁵N and foliar δ¹⁵N, and high isotopic signatures for mycorrhizal fungi. In order to understand the mechanisms creating these patterns, we developed a model of N dynamics and N isotopes (Nitrogen Isotope Fluxes in Terrestrial Ecosystems, NIFTE), which simulated the major transformations of the N cycle and predicted isotopic signatures of different plant species and soil pools. Comparisons with field data from five sites along the successional sequence indicated that NIFTE can duplicate observed patterns in δ¹⁵N of soil, foliage, and mineral N over time. Different scenarios that could account for the observed isotopic patterns were tested in model simulations. Possible mechanisms included increased isotopic fractionation on mineralization, fractionation during the transfer of nitrogen from mycorrhizal fungi to plants, variable fractionation on uptake by mycorrhizal fungi compared to plants, no fractionation on mycorrhizal transfer, and elimination of mycorrhizal fungi as a pool in the model. The model results suggest that fractionation during mineralization must be small (˜2‰), and that no fractionation occurs during plant or mycorrhizal uptake. A net fractionation during mycorrhizal transfer of nitrogen to vegetation provided the best fit to isotopic data on mineral N, plants, soils, and mycorrhizal fungi. The model and field results indicate that the importance of mycorrhizal fungi to N uptake is probably less under conditions of high N availability. Use of this model should encourage a more rigorous assessment of isotopic signatures in ecosystem studies and provide insights into the biologic transformations which affect those signatures. This should lead to an enhanced understanding of some of the fundamental controls on nitrogen dynamics. Received: 1 July 1998 / Accepted: 23 December 1998     

Effects of growth and tissue type on the kinetics of <Superscript>13</Superscript>C and <Superscript>15</Superscript>N incorporation in a rapidly growing ectotherm

The use of stable isotopes to investigate animal diets, habitat use, and trophic level requires understanding the rate at which animals incorporate the ¹³C and ¹⁵N from their diets and the factors that determine the magnitude of the difference in isotopic composition between the animal’s diet and that of its tissues. We determined the contribution of growth and catabolic turnover to the rate of ¹³C and ¹⁵N incorporation into several tissues that can be sampled non-invasively (skin, scute, whole blood, red blood cells, and plasma solutes) in two age classes of a rapidly growing ectotherm (loggerhead turtles, Caretta caretta). We found significant differences in C and N incorporation rates and isotopic discrimination factors (Δ¹³C = δ¹³C_tissues − δ¹³C_diet and Δ¹⁵N = δ¹⁵N_tissues − δ¹⁵N_diet) among tissues and between age classes. Growth explained from 26 to 100% of the total rate of incorporation in hatchling turtles and from 15 to 52% of the total rate of incorporation in juvenile turtles. Because growth contributed significantly to the rate of isotopic incorporation, variation in rates among tissues was lower than reported in previous studies. The contribution of growth can homogenize the rate of isotopic incorporation and limit the application of stable isotopes to identify dietary changes at contrasting time scales and to determine the timing of diet shifts. The isotopic discrimination factor of nitrogen ranged from −0.64 to 1.77‰ in the turtles’ tissues. These values are lower than the commonly assumed average 3.4‰ discrimination factors reported for whole body and muscle isotopic analyses. The increasing reliance on non-invasive and non-destructive sampling in animal isotopic ecology requires that we recognize and understand why different tissues differ in isotopic discrimination factors.     

Contrasts Among Mycorrhizal Plant Guilds in Foliar Nitrogen Concentration and δ15N Along Productivity Gradients of a Boreal Forest

The distribution of plant species in boreal forest understories is hypothesized to reflect mycorrhizal guilds and associated adaptations for organic nitrogen (N) acquisition. In this study of a natural edaphic gradient, where supply rates of inorganic N increase with site productivity, we noted a decline in understory ectomycorrhizal, ericoid, and arbutoid plant communities on productive sites, in contrast to a positive response by most arbuscular species. We then assessed the rate of change in foliar N concentration (N_conc) and abundance of ¹⁵N (δ¹⁵N) of select plants from these mycorrhizal guilds. Two arbuscular plant species (Rubus parviflorus and Viburnum edule) had the sharpest increases in foliar N_conc with enhanced supplies of NH₄ ⁺ and NO₃ ⁻, but with no differences in foliar δ¹⁵N. An ectomycorrhizal species, Abies lasiocarpa, and ericoid species, Vaccinium membranaceum, had parallel increases in both N_conc and δ¹⁵N with soil N supply. The foliar δ¹⁵N of two arbutoid plants (Orthilia secunda and Pyrola asarifolia) were as enriched as ectomycorrhizal sporocarps, likely indicating N transfer from mycorrhizal networks. The depletion of foliar δ¹⁵N by ectomycorrhizal and ericoid plants on poorer sites likely reflected a high degree of N retention and photosynthate demand by fungi, whereas arbuscular plants may have had a less significant δ¹⁵N response because of a more passive role by fungi in scavenging organic N. The results suggest differences in how mycorrhiza exploit diverse soil N supplies (recalcitrant and labile organic N, NH₄ ⁺, NO₃ ⁻, and parasitized N) could be an important factor in boreal plant community composition.     

Carbon nutrition of mature green orchid <Emphasis Type="Italic">Serapias strictiflora</Emphasis> and its mycorrhizal fungus <Emphasis Type="Italic">Epulorhiza</Emphasis> sp.

We studied the nutritional modes of the orchid Serapias strictiflora and its mycorrhizal fungus Epulorhiza sp. using the differences in carbon isotopic composition (δ¹³C) of C₃ orchid and C₄ maize tissues. We found that if cultivated in substrate lacking any organic compounds, the mycorrhizal extraradical mycelia (δ¹³C = −26.3 ± 0.2 ‰) developed well, despite being fully dependent on nutrition from orchid roots (δ¹³C = −28.6 ± 0.1 ‰). If the mycorrhizal fungus had additional access to and colonized decaying maize roots (δ¹³C = −14.6 ± 0.1 ‰), its isotopic composition (δ¹³C = −21.6 ± 0.4 ‰) reflected a mixture of biotrophy and saprotrophy. No statistically significant differences in δ¹³C of new storage tubers were found between Epulorhiza-associated orchids with (δ¹³C = -28.2 ± 0.1 ‰) and without access to maize roots (δ¹³C = −28.6 ± 0.2 ‰). We conclude that autotrophy is the predominant nutritional mode of mature S. strictiflora plants and that they supply their mycorrhizal fungus with substantial amount of carbon (69 ± 3 % of the fungus demand), even if the fungus feeds saprotrophically.     

Insights into nitrogen and carbon dynamics of ectomycorrhizal and saprotrophic fungi from isotopic evidence

The successful use of natural abundances of carbon (C) and nitrogen (N) isotopes in the study of ecosystem dynamics suggests that isotopic measurements could yield new insights into the role of fungi in nitrogen and carbon cycling. Sporocarps of mycorrhizal and saprotrophic fungi, vegetation, and soils were collected in young, deciduous-dominated sites and older, coniferous-dominated sites along a successional sequence at Glacier Bay National Park, Alaska. Mycorrhizal fungi had consistently higher δ¹⁵N and lower δ¹³C values than saprotrophic fungi. Foliar δ¹³C values were always isotopically depleted relative to both fungal types. Foliar δ¹⁵N values were usually, but not always, more depleted than those in saprotrophic fungi, and were consistently more depleted than in mycorrhizal fungi. We hypothesize that an apparent isotopic fractionation by mycorrhizal fungi during the transfer of nitrogen to plants may be attributed to enzymatic reactions within the fungi producing isotopically depleted amino acids, which are subsequently passed on to plant symbionts. An increasing difference between soil mineral nitrogen δ¹⁵N and foliar δ¹⁵N in later succession might therefore be a consequence of greater reliance on mycorrhizal symbionts for nitrogen supply under nitrogen-limited conditions. Carbon signatures of mycorrhizal fungi may be more enriched than those of foliage because the fungi use isotopically enriched photosynthate such as simple sugars, in contrast to the mixture of compounds present in leaves. In addition, some ¹³C fractionation may occur during transport processes from leaves to roots, and during fungal chitin biosynthesis. Stable isotopes have the potential to help clarify the role of fungi in ecosystem processes. Received: 7 January 1998 / Accepted: 9 November 1998     

Influence of functional feeding groups and spatiotemporal variables on the δ<Superscript>15</Superscript>N signature of littoral macroinvertebrates

The δ¹⁵N trophic enrichment in littoral food webs is not well known despite the importance of macroinvertebrates in lacustrine energy fluxes. We wanted to assess the influence of functional feeding group (grazer, collector, shredder, predator, predator–hematophagous, predator–sucker) and spatiotemporal variables (year, month, station of sampling) on littoral macroinvertebrate δ¹⁵N signatures. For 2 years, during the plant growth period phytophilous littoral macroinvertebrates were sampled in Lake St. Pierre, a large fluvial lake of the St. Lawrence River, Québec, Canada. The δ¹⁵N analyses showed that station was the most important factor for explaining δ¹⁵N variation, followed by sampling month and functional feeding group. The organisms sampled in the stations of the south shore, which experienced greater macrophyte abundance, slower currents, and stronger NO₃ depletion exhibited higher δ¹⁵N values than those sampled on the north shore. Grazer-to-predator δ¹⁵N enrichment valued 1.6‰, which is inferior to the 3.4‰ generally admitted in food-web research. Shredders exhibited the lowest δ¹⁵N values and predators–hematophagous the highest. δ¹⁵N signature of invertebrates increased 3‰ through the summer between May and September. Only samples collected within a short period should be pooled to avoid an error value equivalent to one trophic level (1.6) enrichment. Furthermore, it is recommended not to pool macroinvertebrate samples collected at stations with differing watershed land uses.     

First report of the ectomycorrhizal status of boletes on the Northern Yucatan Peninsula, Mexico determined using isotopic methods

Despite their prominent role for tree growth, few studies have examined the occurrence of ectomycorrhizal fungi in lowland, seasonally dry tropical forests (SDTF). Although fruiting bodies of boletes have been observed in a dry tropical forest on the Northern Yucatan Peninsula, Mexico, their occurrence is rare and their mycorrhizal status is uncertain. To determine the trophic status (mycorrhizal vs. saprotrophic) of these boletes, fruiting bodies were collected and isotopically compared to known saprotrophic fungi, foliage, and soil from the same site. Mean δ¹⁵N and δ¹³C values differed significantly between boletes and saprotrophic fungi, with boletes 8.0‰ enriched and 2.5‰ depleted in ¹⁵N and ¹³C, respectively relative to saprotrophic fungi. Foliage was depleted in ¹³C relative to both boletes and saprotrophic fungi. Foliar δ¹⁵N values, on the other hand, were similar to saprotrophic fungi, yet were considerably lower relative to bolete fruiting bodies. Results from this study provide the first isotopic evidence of ectomycorrhizal fungi in lowland SDTF and emphasize the need for further research to better understand the diversity and ecological importance of ectomycorrhizal fungi in these forested ecosystems.     

Vascular plant 15N natural abundance in heath and forest tundra ecosystems is closely correlated with presence and type of mycorrhizal fungi in roots

In this study we show that the natural abundance of the nitrogen isotope 15, δ¹⁵N, of plants in heath tundra and at the tundra-forest ecocline is closely correlated with the presence and type of mycorrhizal association in the plant roots. A total of 56 vascular plant species, 7 moss species, 2 lichens and 6 species of fungi from four heath and forest tundra sites in Greenland, Siberia and Sweden were analysed for δ¹⁵N and N concentration. Roots of vascular plants were examined for mycorrhizal colonization, and the soil organic matter was analysed for δ¹⁵N, N concentration and soil inorganic, dissolved organic and microbial N. No arbuscular mycorrhizal (AM) colonizations were found although potential host plants were present in all sites. The dominant species were either ectomycorrhizal (ECM) or ericoid mycorrhizal (ERI). The δ¹⁵N of ECM or ERI plants was 3.5–7.7‰ lower than that of non-mycorrhizal (NON) species in three of the four sites. This corresponds to the results in our earlier study of mycorrhiza and plant δ¹⁵N which was limited to one heath and one fellfield in N Sweden. Hence, our data suggest that the δ¹⁵N pattern: NON/AM plants > ECM plants ≥ ERI plants is a general phenomenon in ecosystems with nutrient-deficient organogenic soils. In the fourth site, a␣birch forest with a lush herb/shrub understorey, the differences between functional groups were considerably smaller, and only the ERI species differed (by 1.1‰) from the NON species. Plants of all functional groups from this site had nearly twice the leaf N concentration as that found in the same species at the other three sites. It is likely that low inorganic N availability is a prerequisite for strong δ¹⁵N separation among functional groups. Both ECM roots and fruitbodies were ¹⁵N enriched compared to leaves which suggests that the difference in δ¹⁵N between plants with different kinds of mycorrhiza could be due to isotopic fractionation at the␣fungal-plant interface. However, differences in δ¹⁵N between soil N forms absorbed by the plants could also contribute to the wide differences in plant δ¹⁵N found in most heath and forest tundra ecosystems. We hypothesize that during microbial immobilization of soil ammonium the microbial N pool could become ¹⁵N-depleted and the remaining, plant-available soil ammonium ¹⁵N-enriched. The latter could be a main source of N for NON/AM plants which usually have high δ¹⁵N. In contrast, amino acids and other soil organic N compounds presumably are ¹⁵N-depleted, similar to plant litter, and ECM and ERI plants with high uptake of these N forms hence have low leaf δ¹⁵N. Further indications come from the δ¹⁵N of mosses and lichens which was similar to that of ECM plants. Tundra cryptogams (and ECM and ERI plants) have previously been shown to have higher uptake of amino acid than ammonium N; their low δ¹⁵N might therefore reflect the δ¹⁵N of free amino acids in the soil. The concentration of dissolved organic N was 3–16 times higher than that of inorganic N in the sites. Organic nitrogen could be an important N source for ECM and, in particular, ERI plants in heath and forest tundra ecosystems with low release rate of inorganic N from the soil organic matter. Received: 8 June 1997 / Accepted: 28 February 1998     

The carbon and nitrogen isotopic values of particulate organic material from the Florida Keys: a temporal and spatial study

The δ¹⁵N and δ¹³C values of particulate organic material (POM) were analyzed from 35 sites in the Florida Keys over the time interval 2000 to 2002. The sites within the study area were delineated into nine transects stretching from Key West to Key Largo. Each transect consisted of three to five sites extending from close to the Keys to the edge of the reef tract. The POM had mean δ¹⁵N and δ¹³C values of +3.6‰ (σ = ±3.2‰) and −19.9‰ (σ = ±0.6‰) respectively. Over the study period there were no statistically significant changes in δ¹⁵N, δ¹³C, or C:N. For the majority of the sampling dates, the δ¹³C values showed a distinct inshore (δ¹³C = −18.3‰, σ = ±1.0‰) to offshore gradient (δ¹³C = −21.4, σ = ±0.9‰). In contrast, the δ¹⁵N values showed no consistent patterns related to the distance from land. The more positive δ¹³C values of the nearshore samples suggest that the source of the carbon and the nitrogen in the POM in the nearshore was mainly derived from the degradation of seagrass detritus and not from the input of anthropogenically derived material from the Florida Keys. In contrast, the POM on the outer reef was dominated by marine plankton. As mineralization and nitrification of the organic nitrogen pool are major contributors to the dissolved inorganic nitrogen in the water column, it is unlikely that variations in the δ¹⁵N of the algae and other benthic organisms reported in the Florida Keys are related to the input of sewage.     

Differential <Superscript>13</Superscript>C Isotopic Discrimination in Maize at Varying Water Stress and at Low to High Nitrogen Availability

The relationships between ¹³C isotopic discrimination and water stress are well documented for C₃ and C₄ plants. However, the application in the field is hampered by complex interaction patterns with other common stress factors, such as nutrient deficiency. In addition, questions arise if temporal reductions in water availability during crop growth can be traced back using δ¹³C data in the field. The objective of this study therefore is to assess the potential use of δ¹³C observations to quantify water stress and its dynamics in maize (Zea mays L.) grown under low to high nitrogen availability, and to develop tools based on δ¹³C values for its diagnosis in the field. In a pot experiment, carried out in a screen house in Ibadan, Nigeria, we grew maize for 60 days under four watering regimes, (i) optimum (at field capacity) during 60 days, (ii) optimum from 0 to 30 days and stressed (50% field capacity) from 30 to 60 days, (iii) stressed from 0 to 30 days and optimum from 30 to 60 days, and (iv) stressed throughout the 60 days. Nitrogen was applied at three rates (none, moderate (45 kg N ha⁻¹) and high (120 kg N ha⁻¹)). Plants were sampled after 30 and 60 days. At 60 days, leaves developed during the first 30 days were sampled separately from those developed between 30 and 60 days. Shoot production showed a clear water–nitrogen interaction. Nitrogen response increased with decreasing water stress, in particular from 30 to 60 days. δ¹³C values ranged from −12.42‰ to −10.80‰. Overall, a clear and significant water and nitrogen effect (P<0.0001) on the isotopic discrimination in maize was observed, opposite in direction from C₃ plants. δ¹³C values decreased with increasing water stress, but increased with decreasing nitrogen availability, particularly when combined with limited water supply. In addition, isotopic discrimination was observed to be variable within plant, and could be related to a water stress in that growth period, in which the plant parts were developed. This shows that δ¹³C values measured in different plant parts at harvest can be used as a historical account on how water availability varied during the entire cropping cycle.     

Subantarctic Macquarie Island – a model ecosystem for studying animal-derived nitrogen sources using 15N natural abundance

Plants collected from diverse sites on subantarctic Macquarie Island varied by up to 30‰ in their leaf δ¹⁵N values. ¹⁵N natural abundance of plants, soils, animal excrement and atmospheric ammonia suggest that the majority of nitrogen utilised by plants growing in the vicinity of animal colonies or burrows is animal-derived. Plants growing near scavengers and animal higher in the food chain had highly enriched δ¹⁵N values (mean = 12.9‰), reflecting the highly enriched signature of these animals' excrement, while plants growing near nesting penguins and albatross, which have an intermediate food chain position, had less enriched δ¹⁵N values (>6‰). Vegetation in areas affected by rabbits had lower δ¹⁵N values (mean = 1.2‰), while the highly depleted δ¹⁵N values (below −5‰) of plants at upland plateau sites inland of penguin colonies, suggested that a portion of their nitrogen is derived from ammonia (mean ¹⁵N =−10‰) lost during the degradation of penguin guano. Vegetation in a remote area had δ¹⁵N values near −2‰. These results contrast with arctic and subarctic studies that attribute large variations in plant ¹⁵N values to nitrogen partitioning in nitrogen-limited environments. Here, plant ¹⁵N reflects the ¹⁵N of the likely nitrogen sources utilised by plants. Received: 18 December 1997 / Accepted: 13 June 1998     

<Superscript>15</Superscript>N/<Superscript>14</Superscript>N ratios of amino acids as a tool for studying terrestrial food webs: a case study of terrestrial insects (bees,wasps, and hornets)

Compound-specific stable isotope analysis (CSIA) of amino acids is a new method that enables estimates of trophic position for consumers in food webs. We examined the nitrogen isotopic composition (δ¹⁵N) of amino acids of Japanese social insects (three bee, three wasp, and four hornet species) to evaluate the potential of CSIA of amino acids in studies of terrestrial food webs. For wasps, we also examined samples at different growth stages (ranging from egg to adult) to assess the effect of metamorphosis on CSIA estimates of trophic position. The δ¹⁵N values of bulk tissues for Japanese social insects are only weakly correlated with the biologically expected trophic positions. In contrast, the trophic positions estimated from the δ¹⁵N values of amino acids (yielding values of between 2.0 and 2.3 for bees, between 2.8 and 3.3 for wasps, and between 3.5 and 4.1 for hornets) are consistent with the biologically expected trophic positions for these insects (i.e., 2.0 for bees, 3.0 for wasps, and 3.0–4.0 for hornets). Although large variability is observed among the δ¹⁵N values of individual amino acids (e.g., ranging from 3.0 to 14.9‰ for phenylalanine), no significant change is observed in the trophic position during wasp metamorphosis. Thus, the CSIA of amino acids is a powerful tool for investigating not only aquatic food webs but also terrestrial food webs with predatory insects.     

Natural 15N abundance in two nitrogen saturated forest ecosystems

Natural ¹⁵N abundance values were measured in needles, twigs, wood, soil, bulk precipitation, throughfall and soil water in a Douglas fir (Pseudotsuga menziesii (Mirb.) and a Scots pine (Pinus sylvestris L.) stand receiving high loads of nitrogen in throughfall (>50 kg N ha⁻¹ year⁻¹). In the Douglas fir stand δ¹⁵N values of the vegetation ranged between −5.7 and −4.2‰ with little variation between different compartments. The vegetation of the Scots pine stand was less depleted in ¹⁵N and varied from −3.3 to −1.2‰δ¹⁵N. At both sites δ¹⁵N values increased with soil depth, from −5.7‰ and −1.2‰ in the organic layer to +4.1‰ and +4.7‰ at 70 cm soil depth in the Douglas fir and Scots pine stand, respectively. The δ¹⁵N values of inorganic nitrogen in bulk precipitation showed a seasonal variation with a mean in NH₄ ⁺-N of −0.6‰ at the Douglas fir stand and +10.8‰ at the Scots pine stand. In soil water below the organic layer NH₄ ⁺-N was enriched and NO₃ ⁻-N depleted in ¹⁵N, which was interpreted as being caused by isotope fractionation accompanying high nitrification rates in the organic layers. Mean δ¹⁵N values of NH₄ ⁺ and NO₃ ⁻ were very similar in the drainage water at 90 cm soil depth at both sites (−7.1 to −3.8‰). A dynamic N cycling model was used to test the sensitivity of the natural abundance values for the amount of N deposition, the ¹⁵N ratio of atmospheric N deposited and for the intrinsic isotope discrimination factors associated with N transformation processes. Simulated δ¹⁵N values for the N saturated ecosystems appeared particularly sensitive to the ¹⁵N ratio of atmospheric N inputs and discrimination factors during nitrification and mineralization. The N-saturated coniferous forest ecosystems studied were not characterized by elevated natural ¹⁵N abundance values. The results indicated that the natural ¹⁵N abundance values can only be used as indicators for the stage of nitrogen saturation of an ecosystem if the δ¹⁵N values of the deposited N and isotope fractionation factors are taken into consideration. Combining dynamic isotope models and natural ¹⁵N abundance values seems a promising technique for interpreting natural ¹⁵N abundance values found in these forest ecosystems. Received: 5 May 1996 / Accepted: 10 April 1997     

Carbon and nitrogen isotope composition of vegetation on King George Island,maritime Antarctic

We report abundance of ¹³C and ¹⁵N contents in terrestrial plants (mosses, lichens, liverworts, algae and grasses) from the area of Barton Peninsula (King George Island, maritime Antarctic). The investigated plants show a wide range of δ¹³C and δ¹⁵N values between −29.0 and −20.0‰ and between −15.3 and 22.8‰, respectively. The King George Island terrestrial plants show species specificity of both carbon and nitrogen isotope compositions, probably due to differences in plant physiology and biochemistry, related to their sources and in part to water availability. Carbon isotope compositions of Antarctic terrestrial plants are typical of the C₃ photosynthetic pathway. Lichens are characterized by the widest carbon isotope range, from −29.0 to −20.0‰. However, the average δ¹³C value of lichens is the highest (−23.6 ± 2.8‰) among King George Island plants, followed by grasses (−25.6 ± 1.7‰), mosses (−25.9 ± 1.6‰), liverworts (−26.3 ± 0.5‰) and algae (−26.3 ± 1.2‰), partly related to habitats controlled by water availability. The δ¹⁵N values of moss samples range widest (−9.0 to 22.8‰, with an average of 4.6 ± 6.6‰). Lichens are on the average most depleted in ¹⁵N (mean = −7.4 ± 6.4‰), whereas algae are most enriched in ¹⁵N (10.0 ± 3.3‰). The broad range of nitrogen isotope compositions suggest that the N source for these Antarctic terrestrial plants is spatially much variable, with the local presence of seabird colonies being particularly significant.     

Significance of rooting depth in mire plants: Evidence from natural <Superscript>15</Superscript>N abundance

Variation in stable nitrogen isotope ratios (¹⁵N) was assessed for plants comprising two wetland communities, a bog-fen system and a flood plain, in central Japan. ¹⁵N of 12 species from the bog-fen system and six species from the flood plain were remarkably variable, ranging from –5.9 to +1.1 and from +3.1 to +8.7, respectively. Phragmites australis exhibited the highest ¹⁵N value at both sites. Rooting depth also differed greatly with plant species, ranging from 5cm to over 200cm in the bog-fen system. There was a tendency for plants having deeper root systems to exhibit higher ¹⁵N values; plant ¹⁵N was positively associated with rooting depth. Moreover, an increasing gradient of peat ¹⁵N was found along with depth. This evidence, together with the fact that inorganic nitrogen was depleted under a deep-rooted Phragmites australis stand, strongly suggests that deep-rooted plants actually absorb nitrogen from the deep peat layer. Thus, we successfully demonstrated the diverse traits of nitrogen nutrition among mire plants using stable isotope analysis. The ecological significance of deep rooting in mire plants is that it enables those plants to monopolize nutrients in deep substratum layers. This advantage should compensate for any consequential structural and/or physiological costs. Good evidence of the benefits of deep rooting is provided by the fact that Phragmites australis dominates as a tall mire grass.