Large genotypic variation but small variation in N2 fixation among rhizobia nodulating red clover in soils of northern Scandinavia

AIMS: To analyse the symbiotic variations within indigenous populations of rhizobia nodulating red clover (Trifolium pratense L.) in soils of northern Norway and Sweden at different times of the growing season. METHODS AND RESULTS: A total of 431 nodule isolates sampled under field conditions in summer and autumn, were characterized genetically by targeting both chromosomal and symbiotic genes. The Enterobacterial Repetitive Intergenic Consensus polymerase chain reaction (PCR) fingerprinting of chromosomal DNA revealed considerable variation within the isolated populations that was more influenced by geographical origin than sampling time. Analysis of PCR amplified nodEF gene on the symbiotic plasmid by restriction fragment length polymorphism revealed a high proportion of nod types common to the two studied sites. The symbiotic efficiency of the isolates, representing both dominating and rare nodEF genotypes, showed high N(2) fixation rates in symbiosis with the host plant in a greenhouse experiment using the (15)N isotope dilution method. CONCLUSIONS: Effective N(2)-fixing strains of Rhizobium leguminosarum bv. trifolii nodulating red clover are common and genetically diverse in these northern Scandinavia soils. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information on the variability, stability and dynamics of resident populations of rhizobia nodulating red clover in Scandinavian soils which has practical implications for applying biological nitrogen fixation in subarctic plant production.     

Fire effects on ecosystem nitrogen cycling in a Californian bishop pine forest

Fire can cause severe nitrogen (N) losses from grassland, chaparral, and temperate and boreal forest ecosystems. Paradoxically, soil ammonium levels are markedly increased by fire, resulting in high rates of primary production in re-establishing plant communities. In a manipulative experiment, we examined the influence of wild-fire ash residues on soil, microbial and plant N pools in a recently burned Californian bishop pine (Pinus muricata D. Don) forest. Ash stimulated post-fire primary production and ecosystem N retention through direct N inputs from ash to soils, as well as indirect ash effects on soil N availability to plants. These results suggest that redistribution of surface ash after fire by wind or water may cause substantial heterogeneity in soil N availability to plants, and could be an important mechanism contributing to vegetation patchiness in fire-prone ecosystems. In addition, we investigated the impact of fire on ecosystem N cycling by comparing ¹⁵N natural abundance values from recently burned and nearby unburned P. muricata forest communities. At the burned site, ¹⁵N natural abundance in recolonising species was similar to that in bulk soil organic matter. By contrast, there was a marked ¹⁵N depletion in the same species relative to the total soil N pool at the unburned site. These results suggest that plant uptake of nitrate (which tends to be strongly depleted in ¹⁵N because of fractionation during nitrification) is low in recently burned forest communities but could be an important component of eco- system N cycling in mature conifer stands. Received: 29 June 1999 / Accepted: 24 October 1999     

Estimation of symbiotic N2 fixation in an Amazon floodplain forest

Sustainable management for existing Amazonian forests requires an extensive knowledge about the limits of ecosystem nutrient cycles. Therefore, symbiotic nitrogen (N₂) fixation of legumes was investigated in a periodically flooded forest of the central Amazon floodplain (Várzea) over two hydrological cycles (20 months) using the ¹⁵N natural abundance method. No seasonal variation in ¹⁵N abundance (δ ¹⁵N values) in trees which would suggest differences in N₂ fixation rates between the terrestrial and the aquatic phase was found. Estimations of the percentage of N derived from atmosphere (%Ndfa) for the nodulated legumes with Neptunia oleracea on the one side and Teramnus volubilis on the other resulted in mean %Ndfa values between 9 and 66%, respectively. More than half of the nodulated legume species had %Ndfa values above 45%. These relatively high N gains are important for the nodulated legumes during the whole hydrological cycle. With a %Ndfa of 4–5% for the entire Várzea forest, N₂ fixation is important for the ecosystem and therefore, has to be taken into consideration for new sustainable land-use strategies in this area.     

15N natural abundance of vascular rainforest epiphytes: implications for nitrogen source and acquisition

The foliar natural abundance of ¹⁵N was analysed to compare the potential nitrogen sources of vascular rainforest epiphytes and associated soil-rooted trees. Leaves of epiphytes collected from six rainforest communities in Brazil, Australia and the Solomon Islands were depleted in ¹⁵N relative to the trees at each site. Epiphyte δ¹⁵N was as low as -6.4%_o, while trees were generally enriched in ¹⁵N (0.7 to 3.5%_o). These results indicate either that epiphytes use nitrogen sources depleted in ¹⁵N or that discrimination against ¹⁵N is an intrinsic function of epiphyte physiology. At three sites, epiphytes could be grouped into those having both low δ¹⁵N and low leaf-nitrogen content and those possessing both high δ¹⁵N and high leaf-nitrogen content. The second group had δ¹⁵N values in the range sometimes attributable to N₂ fixation (-2 to 0%o). There was no correlation between growth form and δ¹⁵N. It is concluded that epiphytes may utilize ¹⁵N-depleted nitrogen from atmospheric deposition and N₂ fixation.     

Phosphorus availability and elevated CO2 affect biological nitrogen fixation and nutrient fluxes in a clover-dominated sward

The response of biological nitrogen fixation (BNF) to elevated CO(2) was examined in white clover (Trifolium repens)-dominated swards under both high and low phosphorus availability. Mixed swards of clover and buffalo grass (Stenotaphrum secundatum) were grown for 15 months in 0.2 m2 sand-filled mesocosms under two CO2 treatments (ambient and twice ambient) and three nutrient treatments [no N, and either low or high P (5 or 134 kg P ha(-1)); the third nutrient treatment was supplied with high P and N (240 kg N ha(-1))]. Under ambient CO2, high P increased BNF from 410 to 900 kg ha(-1). Elevated CO2 further increased BNF to 1180 kg ha(-1) with high P, but there was no effect of CO2 on BNF with low P. Allocation of N belowground increased by approx. 50% under elevated CO2 irrespective of supplied P. The results suggest that where soil P availability is low, elevated CO2 will not increase BNF, and pasture quality could decrease because of a reduction in aboveground N.     

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.     

Leaf litter inputs reinforce islands of nitrogen fertility in a lowland tropical forest

The role of lowland tropical forest tree communities in shaping soil nutrient cycling has been challenging to elucidate in the face of high species diversity. Previously, we showed that differences in tree species composition and canopy foliar nitrogen (N) concentrations correlated with differences in soil N availability in a mature Costa Rican rainforest. Here, we investigate potential mechanisms explaining this correlation. We used imaging spectroscopy to identify study plots containing 10–20 canopy trees with either high or low mean canopy N relative to the landscape mean. Plots were restricted to an uplifted terrace with relatively uniform parent material and climate. In order to assess whether canopy and soil N could be linked by litterfall inputs, we tracked litter production in the plots and measured rates of litter decay and the carbon and N content of leaf litter and leaf litter leachate. We also compared the abundance of putative N fixing trees and rates of free-living N fixation as well as soil pH, texture, cation exchange capacity, and topographic curvature to assess whether biological N fixation and/or soil properties could account for differences in soil N that were, in turn, imprinted on the canopy. We found no evidence of differences in legume communities, free-living N fixation, or abiotic properties. However, soils beneath high canopy N assemblages received ~ 60% more N via leaf litterfall due to variability in litter N content between plot types. The correlation of N in canopy leaves, leaf litter, and soil suggests that, under similar abiotic conditions, litterfall-mediated feedbacks can help maintain soil N differences among tropical tree assemblages in this diverse tropical forest.

     

Net mineralization of N at deeper soil depths as a potential mechanism for sustained forest production under elevated [CO2]

Elevated atmospheric carbon dioxide concentrations [CO₂] is projected to increase forest production, which could increase ecosystem carbon (C) storage. This study contributes to our broad goal of understanding the causes and consequences of increased fine‐root production and mortality under elevated [CO₂] by examining potential gross nitrogen (N) cycling rates throughout the soil profile. Our study was conducted in a CO₂‐enriched sweetgum (Liquidambar styraciflua L.) plantation in Oak Ridge, TN, USA. We used ¹⁵N isotope pool dilution methodology to measure potential gross N cycling rates in laboratory incubations of soil from four depth increments to 60 cm. Our objectives were twofold: (1) to determine whether N is available for root acquisition in deeper soil and (2) to determine whether elevated [CO₂], which has increased inputs of labile C resulting from greater fine‐root mortality at depth, has altered N cycling rates. Although gross N fluxes declined with soil depth, we found that N is potentially available for roots to access, especially below 15 cm depth where rates of microbial consumption of mineral N were reduced relative to production. Overall, up to 60% of potential gross N mineralization and 100% of potential net N mineralization occurred below 15 cm depth at this site. This finding was supported by in situ measurements from ion‐exchange resins, where total inorganic N availability at 55 cm depth was equal to or greater than N availability at 15 cm depth. While it is likely that trees grown under elevated [CO₂] are accessing a larger pool of inorganic N by mining deeper soil, we found no effect of elevated [CO₂] on potential gross or net N cycling rates. Thus, increased root exploration of the soil volume under elevated [CO₂] may be more important than changes in potential gross N cycling rates in sustaining forest responses to rising atmospheric CO₂.     

Patterns in leaf traits of leguminous and non-leguminous dominant trees along a rainfall gradient in Ghana

Aims Both dominance distribution of species and the composition of the dominant species determine the distribution of traits within community. Leaf carbon (C) and nitrogen (N) isotopic composition are important leaf traits, and such traits of dominant species are associated with ecosystem C, water and N cycling. Very little is known how dominant species with distinct traits (e.g. N-fixing leguminous and non-leguminous trees) mediate resource utilization of the ecosystems in stressful environment.Methods Leaves of 81 dominant leguminous and non-leguminous trees were collected in forest (moist semi-deciduous and dry semi-deciduous ecosystems) and savanna (costal savanna, Guinean savanna and west Sudanian savanna ecosystems) areas and the transitional zone (between the forest and the savanna) along the transect from the south to the north of Ghana. We measured leaf traits, i.e. leaf δ 13 C, leaf δ 15 N, leaf water content, leaf mass per area (LMA) and C and N concentration. Correlation analyses were used to examine trait–trait relationships, and relationships of leaf traits with temperature and precipitation. We used analysis of covariance to test the differences in slopes of the linear regressions between legumes and non-legumes.Important findings Leaf δ 13 C, δ 15 N, leaf water content and LMA did not differ between leguminous and non-leguminous trees. Leaf N concentration and C:N ratio differed between the two groups. Moreover, leaf traits varied significantly among the six ecosystems. δ 13 C values were negatively correlated with annual precipitation and positively correlated with mean annual temperature. In contrast, leaf δ 15 N of non-leguminous trees were positively correlated with annual precipitation and negatively correlated with mean annual temperature. For leguminous trees, such correlations were not significant. We also found significant coordination between leaf traits. However, the slopes of the linear relationships were significantly different between leguminous and non-leguminous trees. Our results indicate that shifts in dominant trees with distinct water-use efficiency were corresponded to the rainfall gradient. Moreover, leguminous trees, those characterized with relative high water-use efficiency in the low rainfall ecosystems, were also corresponded to the relative high N use efficiency. The high proportion of leguminous trees in the savannas is crucial to mitigate nutrient stress.     

Wide geographical and ecological distribution of nitrogen and carbon gains from fungi in pyroloids and monotropoids (Ericaceae) and in orchids

* Stable isotope abundance analyses recently revealed that some European green orchids and pyroloids (Ericaceae) are partially myco-heterotrophic, exploiting mycorrhizal fungi for organic carbon and nitrogen. Here we investigate related species to assess their nutritional mode across various forest and climate types in Germany and California. * C- and N-isotope signatures of five green pyroloids, three green orchids and several obligate myco-heterotrophic species (including the putatively fully myco-heterotrophic Pyrola aphylla) were analysed to quantify the green plants' nutrient gain from their fungal partners and to investigate the constancy of enrichment in (13)C and (15)N of fully myco-heterotrophic plants from diverse taxa and locations relative to neighbouring autotrophic plants. * All green pyroloid and one orchid species showed significant (15)N enrichment, confirming incorporation of fungi-derived N compounds while heterotrophic C gain was detected only under low irradiance in Orthilia secunda. Pyrola aphylla had an isotope signature equivalent to those of fully myco-heterotrophic plants. * It is demonstrated that primarily N gain from mycorrhizal fungi occurred in all taxonomic groups investigated across a wide range of geographical and ecological contexts. The (13)C and (15)N enrichment of obligate myco-heterotrophic plants relative to accompanying autotrophic plants turned out as a fairly constant parameter.     

A distinctive latitudinal trend of nitrogen isotope signature across urban forests in eastern China

Rapid urbanization has greatly altered nitrogen (N) cycling from regional to global scales. Compared to natural forests, urban forests receive much more external N inputs with distinctive abundances of stable N isotope (δ¹⁵N). However, the large-scale pattern of soil δ¹⁵N and its imprint on plant δ¹⁵N remain less well understood in urban forests. By collecting topsoil (0–20 cm) and leaf samples from urban forest patches in nine large cities across a north–south transect in eastern China, we analyzed the latitudinal trends of topsoil C:N ratio and δ¹⁵N as well as the correlations between tree leaf δ¹⁵N and topsoil δ¹⁵N. We further explored the spatial variation of topsoil δ¹⁵N explained by corresponding climatic, edaphic, vegetation-associated, and anthropogenic drivers. Our results showed a significant increase of topsoil C:N ratio towards higher latitudes, suggesting lower N availability at higher latitudes. Topsoil δ¹⁵N also increased significantly at higher latitudes, being opposite to the latitudinal trend of soil N availability. The latitudinal trend of topsoil δ¹⁵N was mainly explained by mean annual temperature, mean annual precipitation, and atmospheric deposition of both ammonium and nitrate. Consequently, tree leaf δ¹⁵N showed significant positive correlations with topsoil δ¹⁵N across all sampled plant species and functional types. Our findings reveal a distinctive latitudinal trend of δ¹⁵N in urban forests and highlight an important role of anthropogenic N sources in shaping the large-scale pattern of urban forest ¹⁵N signature.     

Forest fine-root production and nitrogen use under elevated CO2: contrasting responses in evergreen and deciduous trees explained by a common principle

Despite the importance of nitrogen (N) limitation of forest carbon (C) sequestration at rising atmospheric CO₂ concentration, the mechanisms responsible are not well understood. To elucidate the interactive effects of elevated CO₂ (eCO₂) and soil N availability on forest productivity and C allocation, we hypothesized that (1) trees maximize fitness by allocating N and C to maximize their net growth and (2) that N uptake is controlled by soil N availability and root exploration for soil N. We tested this model using data collected in Free‐Air CO₂ Enrichment sites dominated by evergreen (Pinus taeda; Duke Forest) and deciduous [Liquidambar styraciflua; Oak Ridge National Laboratory (ORNL)] trees. The model explained 80–95% of variation in productivity and N‐uptake data among eCO₂, N fertilization and control treatments over 6 years. The model explains why fine‐root production increased, and why N uptake increased despite reduced soil N availability under eCO₂ at ORNL and Duke. In agreement with observations at other sites, the model predicts that soil N availability reduced below a critical level diminishes all eCO₂ responses. At Duke, a negative feedback between reduced soil N availability and N uptake prevented progressive reduction in soil N availability at eCO₂. At ORNL, soil N availability progressively decreased because it did not trigger reductions in N uptake; N uptake was maintained at ORNL through a large increase in the production of fast turnover fine roots. This implies that species with fast root turnover could be more prone to progressive N limitation of carbon sequestration in woody biomass than species with slow root turnover, such as evergreens. However, longer term data are necessary for a thorough evaluation of this hypothesis. The success of the model suggests that the principle of maximization of net growth to control growth and allocation could serve as a basis for simplification and generalization of larger scale forest and ecosystem models, for example by removing the need to specify parameters for relative foliage/stem/root allocation.     

Effects of inorganic nitrogen and phosphorus supply on nitrogen aquisition from alanine by birch seedlings in symbiosis with Paxillus involutus

Northern forests are exposed to relatively high ammonia inputs due to high atmospheric deposition and the common practise of forest fertilization. It is not known how increased soil NH₄ ⁺concentrations affect acquisition of symbiosis-mediated N from organic sources. We examined the effect of inorganic N and P availability on N acquisition from alanine by 43 weeks old birch (Betula pendula) seedlings in symbiosis with the ectomycorrhizal fungus Paxillus involutus. The seedlings were exposed for 9 weeks to nutrient additions equivalent to 43 kg N and 6.4 kg P ha^-1 (low N and P availability), 250 kg N and 38 kg P ha^-1(high N and P availability) or to 250 kg N and 6.4 kg P ha^-1 (high N and low P availability). Carbon and nitrogen allocation between the symbionts was assessed by exposing the foliage to ¹⁴CO₂ and the mycelium to ¹⁵N-alanine or ¹⁵NH₄ ⁺ simultaneously and measuring the distribution of the isotopic tracers after a three-day chase period. High inorganic N combined with low P availability did not have marked effect on symbiosis-mediated N uptake from alanine, whilst high N and P availability reduced alanine-derived ¹⁵N translocation by the fungus to the plant. Shoot ¹⁵N concentration and concentration of ¹⁴C in the extramatrical mycelium correlated significantly across treatments pointing to controlled reciprocity of transactions between the partners.     

Nutrient Limitation to Nitrogen Fixation in Young Volcanic Sites

I used measures of ¹⁵N natural abundance and of nitrogenase activity (acetylene reduction) to examine whether the supply of non-N nutrients limits rates of N₂ fixation on young volcanic substrates in Hawaii. Leaves of the dominant tree (Metrosideros polymorpha, a nonfixer) were strongly depleted in ¹⁵N in control plots (–10.8 to –11.1⁰/₀₀). More than 5 y of repeated fertilization with P increased δ¹⁵N to –8.9 to –9.9⁰/₀₀, and the addition of all other essential plant nutrients (except N) together with P further increased ¹⁵N to –8.1 to –9.3⁰/₀₀. This pattern is consistent with enhanced N₂ fixation, because newly fixed N would have a δ¹⁵N near 0⁰/₀₀. Assays of nitrogenase activity in the experimental plots demonstrated that potential N fixation associated with nonvascular plants and with tree and fern litter were increased significantly by additions of P and by the combined nutrient treatment; when these were added together, the increase in nitrogenase activity was 6- to 11-fold over control plots. The supply of P and other weathering-derived nutrients constrains rates of N₂ fixation in these young volcanic sites and thereby contributes to the maintenance of N limitation to primary production and other ecosystem processes. Received 7 January 1999; accepted 3 May 1999.     

The contribution of nitrogen by promiscuous soybeans to maize based cropping the moist savanna of Nigeria

Agronomic results indicate that maize grain yields generally are higher when the crop is planted following soybean than in continuous maize cultivation in the moist savanna agroecological zones of West Africa. Many factors have been hypothesized to explain this phenomenon, including enhanced N availability and the so-called `rotational effect'. There is, however, hardly any quantitative information on the residual N benefits of promiscuous soybeans to subsequent cereal crops grown in rotation with soybean. Three IITA promiscuous soybean breeding lines and two Brazilian soybean lines were grown in 1994 and 1995 at Mokwa in the southern Guinea savanna, Nigeria, to quantify the nitrogen contribution by soybeans to a succeeding crop of maize grown in rotation with soybean for two consecutive years, 1996 and 1997 using two methods of introducing ¹⁵N into soil (fresh ¹⁵N labelling and its residual ¹⁵N) and three maize cultivars (including one cultivar with high N use efficiency) used as reference plants. The nodulating soybeans fixed between 44 and 103 kg N ha^–1 of their total N and had an estimated net N balance input from fixation following grain harvest ranging from –8 to 43 kg N ha^–1. Results in 1996 and in 1997 showed that maize growing after soybean had significantly higher grain yield (1.2 – 2.3-fold increase compared to maize control) except for maize cultivar Oba super 2 (8644-27) (a N-efficient hybrid). The ¹⁵N isotope dilution method was able to estimate N contribution by promiscuous soybeans to maize only in the first succeeding maize crop grown in 1996 but not in the second maize crop in 1997. The first crop of maize grown after soybean accumulated an average between 10 and 22 kg N ha^–1 from soybean residue, representing 17–33% of the soybean total N ha^–1. The percentage ¹⁵N derived from residue recovery in maize grown after maize was influenced by the maize cultivars. Maize crop grown after the N-efficient hybrid cultivar Oba Super 2 (844-27) had similar ¹⁵N values similar to maize grown after soybeans, confirming the ability of this cultivar to use N efficiently in low N soil due to an efficient N translocation ability. The maize crop in 1997 grown after maize had lower ¹⁵N enrichment than that grown in soybean plots, suggesting that soybean residues contributed a little to soil available N and to crop N uptake by the second maize crop. The differential mineralization and immobilization turnover of maize and soybean residues in these soils may be important and N contribution estimates in longer term rotation involving legumes and cereals may be difficult to quantify using the ¹⁵N labelling approaches. Therefore alternative methods are required to measure N release from organic residues in these cropping systems.     

Growth and nitrogen accretion of dinitrogen-fixing Alnus glutinosa (L.) Gaertn. under elevated carbon dioxide

Short-term studies of tree growth at elevated CO₂ suggest that forest productivity may increase as atmospheric CO₂ concentrations rise, although low soil N availability may limit the magnitude of this response. There have been few studies of growth and N₂ fixation by symbiotic N₂-fixing woody species under elevated CO₂ and the N inputs these plants could provide to forest ecosystems in the future. We investigated the effect of twice ambient CO₂ on growth, tissue N accretion, and N₂ fixation of nodulated Alnus glutinosa (L.) Gaertn. grown under low soil N conditions for 160 d. Root, nodule, stem, and leaf dry weight (DW) and N accretion increased significantly in response to elevated CO₂. Whole-plant biomass and N accretion increased 54% and 40%, respectively. Delta-¹⁵N analysis of leaf tissue indicated that plants from both treatments derived similar proportions of their total N from symbiotic fixation suggesting that elevated CO₂ grown plants fixed approximately 40% more N than did ambient CO₂ grown plants. Leaves from both CO₂ treatments showed similar relative declines in leaf N content prior to autumnal leaf abscission, but total N in leaf litter increased 24% in elevated compared to ambient CO₂ grown plants. These results suggest that with rising atmospheric CO₂ N₂-fixing woody species will accumulate greater amounts of biomass N through N₂ fixation and may enhance soil N levels by increased litter N inputs.     

Decomposition of <Superscript>15</Superscript>N-labelled beech litter and fate of nitrogen derived from litter in a beech forest

The decomposition and the fate of ¹⁵N- labelled beech litter was monitored in a beech forest (Vosges mountains, France) over 3 years. Circular plots around beech trees were isolated from neighbouring tree roots by soil trenching. After removal of the litter layer, ¹⁵N-labelled litter was distributed on the soil. Samples [labelled litter, soil (0–15 cm depths], fine roots, mycorrhizal root tips, leaves) were collected during the subsequent vegetation periods and analysed for total N and ¹⁵N concentration. Mass loss of the ¹⁵N-labelled litter was estimated using mass loss data from a litterbag experiment set up at the field site. An initial and rapid release of soluble N from the decomposing litter was balanced by the incorporation of exogenous N into the litter. Fungal N accounted for approximately 35% of the N incorporation. Over 2 years, litter N was continuously released and rates of N and mass loss were equivalent, while litter N was preferentially lost during the 3rd year. Released ¹⁵N accumulated essentially at the soil surface. ¹⁵N from the decomposing litter was rapidly (i.e. in 6 months) detected in roots and beech leaves and its level increased regularly and linearly over the course of the labelling experiment. After 3 years, about 2% of the original litter N had accumulated in the trees. ¹⁵N budgets indicated that soluble N was the main source for soil microbial biomass. Nitrogen accumulated in storage compounds was the main source of leaf N, while soil organic N was the main source of mycorrhizal N. Use of ¹⁵N-labelled beech litter as decomposing substrate allowed assessment of the fate of litter N in the soil and tree N pools in a beech forest on different time scales. Received: 3 May 1999 / Accepted: 3 January 2000     

Nitrogen transfer from nodulating soybean to maize or to nonnodulating soybean in intercrops: the 15N dilution method

In 1985, 1986 and 1988, maize (Zea mays L.) was monocropped or intercropped with nodulating or nonnodulating soybean (Glycine max [L.] Merr.). In addition, nodulating soybean and nonnodulating soybean were each monocropped and grown as a mixture. In 1985 and 1986, treatments were grown at 0 and 60 kg N ha^–1 and in 1988, the treatments were grown without N fertilizer, on N-depeted soil and on non-N-depleted soil. ¹⁵N enriched N was applied to soil in all the aforementioned treatments to test for N transfer from nodulating soybean to non-N₂-fixing crops by the ¹⁵N dilution method.The ¹⁵N dilution method did not show the occurrence of N transfer in 1985 and 1986, but the N sparing effect was evident from the total N uptake of nonnodulating soybean, dwarf maize and tall maize, in 1986. In 1988, maize and nonnodulating soybean seed yields and seed N yields were higher on non-N-depleted soil than on N-depleted soil. On N-depleted soil, the ¹⁵N dilution method indicated N transfer from nodulating soybean to maize and to nonndulating soybean. At a population ratio of 67% nodulating soybean to 33% nonnodulating soybean, N transfer was also seen on non-N-depleted soil in 1988.     

Response to inoculation and N fertilization for increased yield and biological nitrogen fixation of common bean (Phaseolus vulgaris L.)

Common bean (Phaseolus vulgaris L.) is able to fix 20–60 kg N ha^–1 under tropical environments in Brazil, but these amounts are inadequate to meet the N requirement for economically attractive seed yields. When the plant is supplemented with N fertilizer, N₂ fixation by Rhizobium can be suppressed even at low rates of N. Using the ¹⁵N enriched method, two field experiments were conducted to compare the effect of foliar and soil applications of N-urea on N₂ fixation traits and seed yield. All treatments received a similar fertilization including 10 kg N ha^–1 at sowing. Increasing rates of N (10, 30 and 50 kg N ha^–1) were applied for both methods. Foliar application significantly enhanced nodulation, N₂ fixation (acetylene reduction activity) and yield at low N level (10 kg N ha^–1). Foliar nitrogen was less suppressive to nodulation, even at higher N levels, than soil N treatments. In the site where established Rhizobium was in low numbers, inoculation contributed substantially to increased N₂ fixation traits and yield. Both foliar and soil methods inhibited nodulation at high N rates and did not significantly increase bean yield, when comparing low (10 kg N ha^–1) and high (50 kg N ha^–1) rates applied after emergence. In both experiments, up to 30 kg N ha^–1 of biologically fixed N₂ were obtained when low rates of N were applied onto the leaves.     

利用15N示踪技术研究木荷与马尾松幼苗叶片对NO2的吸收与分配

大气氮氧化物(NO_x=NO+NO₂)随着干沉降进入森林生态系统时,会首先接触森林冠层。森林乔木能通过叶片吸收多少NO₂以及对吸收的NO₂是如何分配的,目前尚不清楚。该研究利用¹⁵N稳定同位素示踪技术,对中国南方常见乔木树种木荷(Schima superba)和马尾松(Pinus massoniana)幼苗在黑暗和光照两种条件下进行了¹⁵NO₂静态箱熏蒸实验,检测并分析了两种植物的¹⁵N回收率以及吸收的NO₂在植物各组织中的分配结果。结果显示:植物主要通过气孔吸收NO₂,木荷和马尾松在黑暗条件下整体分别能回收10.3%±5.9%和20.4%±7.0%¹⁵NO₂,在光照条件下整体分别能回收35.9%±5.4%和68.2%±7.6%¹⁵NO₂。两种植物各组织中的平均干质量¹⁵