Functional diversity in an Amazonian rainforest of French Guyana: a dual isotope approach (δ15N and δ13C)

Functional aspects of biodiversity were investigated in a lowland tropical rainforest in French Guyana (5°2′N, annual precipitation 2200 mm). We assessed leaf δ¹⁵N as a presumptive indicator of symbiotic N₂ fixation, and leaf and wood cellulose δ¹³C as an indicator of leaf intrinsic water-use efficiency (CO₂ assimilation rate/leaf conductance for water vapour) in dominant trees of 21 species selected for their representativeness in the forest cover, their ecological strategy (pioneers or late successional stage species, shade tolerance) or their potential ability for N₂ fixation. Similar measurements were made in trees of native species growing in a nearby plantation after severe perturbation (clear cutting, mechanical soil disturbance). Bulk soil δ¹⁵N was spatially quite uniform in the forest (range 3–5‰), whereas average leaf δ¹⁵N ranged from −0.3‰ to 3.5‰ in the different species. Three species only, Diplotropis purpurea, Recordoxylon speciosum (Fabaceae), and Sclerolobium melinonii (Caesalpiniaceae), had root bacterial nodules, which was also associated with leaf N concentrations higher than 20 mg g⁻¹. Although nodulated trees displayed significantly lower leaf δ¹⁵N values than non-nodulated trees, leaf δ¹⁵N did not prove a straightforward indicator of symbiotic fixation, since there was a clear overlap of δ¹⁵N values for nodulated and non-nodulated species at the lower end of the δ¹⁵N range. Perturbation did not markedly affect the difference δ¹⁵N_soil−δ¹⁵N_leaf, and thus the isotopic data provide no evidence of an alteration in the different N acquisition patterns. Extremely large interspecific differences in sunlit leaf δ¹³C were observed in the forest (average values from −31.4 to −26.7‰), corresponding to intrinsic water-use efficiencies (ratio CO₂ assimilation rate/leaf conductance for water vapour) varying over a threefold range. Wood cellulose δ¹³C was positively related to total leaf δ¹³C, the former values being 2–3‰ higher than the latter ones. Leaf δ¹³C was not related to leaf δ¹⁵N at either intraspecific or interspecific levels. δ¹³C of sunlit leaves was highest in shade hemitolerant emergent species and was lower in heliophilic, but also in shade-tolerant species. For a given species, leaf δ¹³C did not differ between the pristine forest and the disturbed plantation conditions. Our results are not in accord with the concept of existence of functional types of species characterized by common suites of traits underlying niche differentiation; rather, they support the hypothesis that each trait leads to a separate grouping of species. Received: 18 August 1997 / Accepted: 14 April 1998     

Feeding habits of Hymenoptera and Isoptera in a tropical rain forest as revealed by nitrogen and carbon isotope ratios

Despite the recognition of the functional role of Hymenoptera (ants, bees and wasps) and Isoptera (termites) in tropical ecosystems, their detailed feeding habits are not well known. To examine the feeding habits of these groups, we measured nitrogen (N) and carbon (C) stable isotope ratios (δ¹⁵N and δ¹³C) of hymenopterans (12 families, ≥16 genera and ≥32 species) and isopterans (one family and 10 species) collected in a tropical rain forest, Sarawak, Malaysia. We compared the isotopic signatures of these insects to those previously reported for other consumers collected in the same forest. The δ¹⁵N and δ¹³C values of these insects overlapped with those of the other consumers, indicating that they have access to diverse C and N sources in the forest. The δ¹⁵N values of ants and termites indicated that their feeding habits range along a continuum from herbivory (i.e. dependent on honeydew and nectar) to predation and from wood-feeders to soil-feeders, respectively. In addition, the δ¹⁵N values of wasps varied greatly from −0.1‰ (Braconidae sp.) to 8.6‰ (Bembix sp.), suggesting that their feeding habits also range from omnivory to predation. The ant species Camponotus gigas had δ¹³C values similar to those of invertebrate detritivores and omnivores rather than to those of invertebrate herbivores, although the diet of this species consists mostly of honeydew. This discrepancy suggests that the ant uses carbohydrates as an energy source, the isotopic signatures of which are not well retained in the body tissues. Values of both δ¹⁵N and δ¹³C of the predatory army ant Leptogenys diminuta and the soil-feeding termite Dicuspiditermes nemorosus did not differ significantly, indicating that both trophic level and the humification of feeding substrates can increase the isotopic signatures of terrestrial consumers.     

Nitrogen availability patterns in white-sand vegetations of Central Brazilian Amazon

Addressing spatial variability in nitrogen (N) availability in the Central Brazilian Amazon, we hypothesized that N availability varies among white-sand vegetation types (campina and campinarana) and lowland tropical forests (dense terra-firme forests) in the Central Brazilian Amazon, under the same climate conditions. Accordingly, we measured soil and foliar N concentration and N isotope ratios (δ¹⁵N) throughout the campina-campinarana transect and compared to published dense terra-firme forest results. There were no differences between white-sand vegetation types in regard to soil N concentration, C:N ratio and δ¹⁵N across the transect. Both white-sand vegetation types showed very low foliar N concentrations and elevated foliar C:N ratios, and no significant difference between site types was observed. Foliar δ¹⁵N was depleted, varying from −9.6 to 1.6‰ in the white-sand vegetations. The legume Aldina heterophylla had the highest average δ¹⁵N values (−1.5‰) as well as the highest foliar N concentration (2.1%) while the non-legume species had more depleted δ¹⁵N values and the average foliar N concentrations varied from 0.9 to 1.5% among them. Despite the high variation in foliar δ¹⁵N among plants, a significant and gradual ¹⁵N-enrichment in foliar isotopic signatures throughout the campina–campinarana transect was observed. Individual plants growing in the campinarana were significantly enriched in ¹⁵N compared to those in campina. In the white-sand N-limited ecosystems, the differentiation of N use seems to be a major cause of variations observed in foliar δ¹⁵N values throughout the campina–campinarana transect.     

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.     

Nitrogen stable isotopic composition of leaves and soil: Tropical versus temperate forests

Several lines of evidence suggest that nitrogen in most tropical forests is relatively more available than N in most temperate forests, and even that it may function as an excess nutrient in many tropical forests. If this is correct, tropical forests should have more open N cycles than temperate forests, with both inputs and outputs of N large relative to N cycling within systems. Consequent differences in both the magnitude and the pathways of N loss imply that tropical forests should in general be more¹⁵N enriched than are most temperate forests. In order to test this hypothesis, we compared the nitrogen stable isotopic composition of tree leaves and soils from a variety of tropical and temperate forests. Foliar ¹⁵N values from tropical forests averaged 6.5 higher than from temperate forests. Within the tropics, ecosystems with relatively low N availability (montane forests, forests on sandy soils) were significantly more depleted in¹⁵N than other tropical forests. The average ¹⁵N values for tropical forest soils, either for surface or for depth samples, were almost 8 higher than temperate forest soils. These results provide another line of evidence that N is relatively abundant in many tropical forest ecosystems.     

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     

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     

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     

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.     

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.     

Grain 15N of crops applied with organic and chemical fertilizers in a four-year rotation

Variations in crop grain and soil N isotope composition (δ¹⁵N) in relation to liquid hog manure (δ¹⁵N of total N was +5.1‰), solid cattle manure (+7.9‰) and chemical fertilizer (+0.7‰ for urea and −1.9‰ for ammonium phosphate) applications, and control (no fertilizer application) were examined through a 4-year crop rotation under field conditions. Canola (Brassica napus), hull-less barley (Hordeum vulgare), wheat (Triticum aestivum), and canola were grown sequentially from 2000 (year 1) to 2003 (year 4). From year 2, hog manure or chemical fertilizers, but not cattle manure, treatments increased grain N concentrations over the control. Grain δ¹⁵N (+0.3 to +2.5‰) of crops applied with chemical fertilizers was lower than those in the other treatments, reflecting the effects of the N source with a lower δ¹⁵N, while the manure treatments tended to increase grain δ¹⁵N. The higher grain δ¹⁵N of crops applied with hog manure (+5.6 to +8.4‰) than those applied with cattle manure (+2.2 to +4.1‰) reflected the higher N availability of liquid hog manure (up to 70% as NH ₄ ⁺ ) than solid cattle manure (99% organic N) and higher potentials for ammonia volatilization loss in hog manure rather than differences in manure δ¹⁵N signatures. Soil total- and extractable-N concentrations and δ¹⁵N tended to vary with the application of N sources with different N isotope composition and availability. Our study expanded the application of the δ¹⁵N technique for detecting N source (organic vs chemical) effects on N isotopic composition to field conditions and across a 4-year rotation, and revealed that N availability played a greater role than the δ¹⁵N signature of N sources in determining crop δ¹⁵N under the studied conditions. Section Editor: H. Lambers     

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.     

Foliar and fungal <Superscript>15</Superscript> N:<Superscript>14</Superscript> N ratios reflect development of mycorrhizae and nitrogen supply during primary succession: testing analytical models

Nitrogen isotopes (¹⁵N/¹⁴N ratios, expressed as δ¹⁵N values) are useful markers of the mycorrhizal role in plant nitrogen supply because discrimination against ¹⁵N during creation of transfer compounds within mycorrhizal fungi decreases the ¹⁵N/¹⁴N in plants (low δ¹⁵N) and increases the ¹⁵N/¹⁴N of the fungi (high δ¹⁵N). Analytical models of ¹⁵N distribution would be helpful in interpreting δ¹⁵N patterns in fungi and plants. To compare different analytical models, we measured nitrogen isotope patterns in soils, saprotrophic fungi, ectomycorrhizal fungi, and plants with different mycorrhizal habits on a glacier foreland exposed during the last 100 years of glacial retreat and on adjacent non-glaciated terrain. Since plants during early primary succession may have only limited access to propagules of mycorrhizal fungi, we hypothesized that mycorrhizal plants would initially be similar to nonmycorrhizal plants in δ¹⁵N and then decrease, if mycorrhizal colonization were an important factor influencing plant δ¹⁵N. As hypothesized, plants with different mycorrhizal habits initially showed similar δ¹⁵N values (−4 to −6‰ relative to the standard of atmospheric N₂ at 0‰), corresponding to low mycorrhizal colonization in all plant species and an absence of ectomycorrhizal sporocarps. In later successional stages where ectomycorrhizal sporocarps were present, most ectomycorrhizal and ericoid mycorrhizal plants declined by 5–6‰ in δ¹⁵N, suggesting transfer of ¹⁵N-depleted N from fungi to plants. The values recorded (−8 to −11‰) are among the lowest yet observed in vascular plants. In contrast, the δ¹⁵N of nonmycorrhizal plants and arbuscular mycorrhizal plants declined only slightly or not at all. On the forefront, most ectomycorrhizal and saprotrophic fungi were similar in δ¹⁵N (−1 to −3‰), but the host-specific ectomycorrhizal fungus Cortinarius tenebricus had values of up to 7‰. Plants, fungi and soil were at least 4‰ higher in δ¹⁵N from the mature site than in recently exposed sites. On both the forefront and the mature site, host-specific ectomycorrhizal fungi had higher δ¹⁵N values than ectomycorrhizal fungi with a broad host range. From these isotopic patterns, we conclude:(1) large enrichments in ¹⁵N of many ectomycorrhizal fungi relative to co-occurring ectomycorrhizal plants are best explained by treating the plant-fungal-soil system as a closed system with a discrimination against ¹⁵N of 8–10‰ during transfer from fungi to plants, (2) based on models of ¹⁵N mass balance, ericoid and ectomycorrhizal fungi retain up to two-thirds of the N in the plant-mycorrhizal system under the N-limited conditions at forefront sites, (3) sporocarps are probably enriched in ¹⁵N by an additional 3‰ relative to available nitrogen, and (4) host-specific ectomycorrhizal fungi may transfer more N to plant hosts than non-host-specific ectomycorrhizal fungi. Our study confirms that nitrogen isotopes are a powerful tool for probing nitrogen dynamics between mycorrhizal fungi and associated plants.     

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.     

Changes in stable isotopic signatures of soil nitrogen and carbon during 40 years of forest development

Understanding what governs patterns of soil δ¹⁵N and δ¹³C is limited by the absence of these data assembled throughout the development of individual ecosystems. These patterns are important because stable isotopes of soil organic N and C are integrative indicators of biogeochemical processing of soil organic matter. We examined δ¹⁵N of soil organic matter (δ¹⁵N_SOM) and δ¹³C_SOM of archived soil samples across four decades from four depths of an aggrading forest in southeastern USA. The site supports an old-field pine forest in which the N cycle is affected by former agricultural fertilization, massive accumulation of soil N by aggrading trees over four decades, and small to insignificant fluxes of N via NH₃ volatilization, nitrification, and denitrification. We examine isotopic data and the N and C dynamics of this ecosystem to evaluate mechanisms driving isotopic shifts over time. With forest development, δ¹³C_SOM became depth-dependent. This trend resulted from a decline of ~2‰ in the surficial 15 cm of mineral soil to −26.0‰, due to organic matter inputs from forest vegetation. Deeper layers exhibited relatively little trend in δ¹³C_SOM with time. In contrast, δ¹⁵N_SOM was most dynamic in deeper layers. During the four decades of forest development, the deepest layer (35–60 cm) reached a maximum δ¹⁵N value of 9.1‰, increasing by 7.6‰. The transfer of >800 kg ha⁻¹ of soil organic N into aggrading vegetation and the forest floor and the apparent large proportion of ectomycorrhizal (ECM) fungi in these soils suggest that fractionation via microbial transformations must be the major process changing δ¹⁵N in these soils. Accretion of isotopically enriched compounds derived from microbial cells (i.e., ECM fungi) likely promote isotopic enrichment of soils over time. The work indicates the rapid rate at which ecosystem development can impart δ¹⁵N_SOM and δ¹³C_SOM signatures associated with undisturbed soil profiles.     

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.     

Contributions of stable-isotope data to elucidating food webs of Mediterranean rocky littoral fishes

The food webs of rocky infra-littoral ecosystems in the Mediterranean have been little studied. In this investigation stable isotopes and dietary data were compared in an attempt to describe features of the food webs concerned. δ¹³C and δ¹⁵N were determined for plants, invertebrates and fishes from the Bay of Calvi, Corsica. Dietary data were derived from the literature. δ¹³C of plants ranged from –8.59‰ to –33.74‰, of benthic invertebrates from –17.0‰ to –20.52‰, of planktonic invertebrates from –20.08‰ to –22.34‰ and of fishes from –16.27‰ to –19.59‰. δ¹⁵N was generally greater at higher trophic levels. δ¹⁵N of plants was 0.95–2.92‰, of benthic invertebrates 1.69–6.54‰, of planktonic invertebrates 3.51–6.82‰ and of fishes 4.63–9.77‰. ¹³C enrichment tended to be associated with benthic food chains and ¹³C depletion with planktonic chains. Stable-isotope data suggested more varied diets for many species than implied by gut-contents data. Omnivory and trophic plasticity were widespread, and many consumers fed lower down the food chain than previous studies had suggested. Both stable-isotope and gut-contents analysis resolved differences between fishes feeding on planktonic and benthic prey and indicated that the herbivorous fish Sarpa salpa fed on a diet substantially different from that of other fishes. Zooplankton were important in the diets of several consumers (both primary and secondary), as was plankton derived detritus. One species of fish previously identified as planktivorous was shown to feed largely on benthic organisms, whilst several species of benthic invertebrates may feed on plankton-derived detritus. Although herbivores seemed to obtain most of their C from macroalgae, δ¹⁵N data suggested that many of these animals supplemented their intake of N, although gut-contents analysis did not provide evidence for such uptake. The isotopic data have elucidated several features of the food web which we would not otherwise have detected. Received: 26 April 1999 / Accepted: 24 September 1999     

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     

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.     

Estimating the uptake of traffic-derived NO2 from 15N abundance in Norway spruce needles

The ¹⁵N ratio of nitrogen oxides (NO_x) emitted from vehicles, measured in the air adjacent to a highway in the Swiss Middle Land, was very high [δ¹⁵N(NO₂) = +5.7‰]. This high ¹⁵N abundance was used to estimate long-term NO₂ dry deposition into a forest ecosystem by measuring δ¹⁵N in the needles and the soil of potted and autochthonous spruce trees [Picea abies (L.) Karst] exposed to NO₂ in a transect orthogonal to the highway. δ¹⁵N in the current-year needles of potted trees was 2.0‰ higher than that of the control after 4 months of exposure close to the highway, suggesting a 25% contribution to the N-nutrition of these needles. Needle fall into the pots was prevented by grids placed above the soil, while the continuous decomposition of needle litter below the autochthonous trees over previous years has increased δ¹⁵N values in the soil, resulting in parallel gradients of δ¹⁵N in soil and needles with distance from the highway. Estimates of NO₂ uptake into needles obtained from the δ¹⁵N data were significantly correlated with the inputs calculated with a shoot gas exchange model based on a parameterisation widely used in deposition modelling. Therefore, we provide an indication of estimated N inputs to forest ecosystems via dry deposition of NO₂ at the receptor level under field conditions. Received: 7 November 1997 / Accepted: 16 September 1998