Belowground competitive suppression of seedling growth by grass in an African savanna

Coexistence of N₂-fixing legumes and non-legume trees with grasses in African savannas results in intense competition between these life-forms. We hypothesised that belowground competition might induce different nutritional constraints in N₂- versus non-N₂-fixing species. A field (Hluhluwe-imFolozi nature reserve, South Africa) competition experiment with two N₂-fixing legume species (Acacia burkei and Acacia karroo) and two non-N₂-fixing species (Schotia brachypetala and Spirostachys africana) both with clipped grass and without grass was established. Plants were supplied with no fertilizer, or generous amounts of fertilizer (200?kg?N?ha^?1, 100?kg P₂O₅ ha^?1, 7.1?kg K₂O ha^?1) supplied as either 28?C10 (N?CK), P or a combination of these fertilizers (NPK). Regularly clipped grass suppressed growth (by more than 90?%) of both N₂- and non-N₂-fixing seedlings equally. Biomass accumulation of seedlings grown with grass and the grasses themselves responded positively to NK and/or NPK, but not P, although P-fertilization did have effects on foliar [N] and ??¹⁵N values of trees and grasses showing that plants accessed the fertilizer. Tree ??¹⁵N values and foliar [N] were also modified by NPK, demonstrating access to fertilizer. However, the ameliorative effects of NPK on grass competition-induced biomass suppression were only partial. This may be due to ??non-resource competition?? (i.e. root gaps) imposed by dense grass roots. The fact that nutrients were able to partially ameliorate the effects of grass competition, however, indicates that such ??non-resource competition?? may be partially overcome by even more generous supply of fertilizers.     

Stable carbon and nitrogen isotope ratios of Eucalyptus and Acacia species along a seasonal rainfall gradient in Western Australia

Key message

Eucalyptus and Acacia species were surprisingly similar with respect to variations in δ ¹³ C, δ ¹⁵ N. Both genera respond with speciation and associated changes in leaf structure to drought.

Abstract

Stable carbon and nitrogen isotope ratios (δ¹³C and δ¹⁵N) in leaves of eucalypts (Corymbia and Eucalyptus) and Acacia (and some additional Fabaceae) species were investigated together with specific leaf area (SLA), leaf nitrogen (N) and leaf phosphorous (P) concentration along a north–south transect through Western Australia covering winter- and summer-dominated rainfall between 100 and 1,200 mm annually. We investigated 62 eucalypts and 78 woody Fabaceae species, mainly of the genus Acacia. Leaf δ¹³C values of Eucalyptus and Acacia species generally increased linearly with latitude from ?29.5 ± 1.3 ‰ in the summer-dominated rainfall zone (15°S–18°S) to about ?25.7 ± 1.1 ‰ in the winter-dominated rainfall zone (29°S–31°S). δ¹⁵N increased initially with southern latitudes (0.5 ± 1.6 ‰ at 15°S; 5.8 ± 3.3 ‰ at 24–29°S) but decreased again further South (4.6 ± 3.5 ‰ at 31°S). The variation in δ¹³C and δ¹⁵N was probably due to speciation of Eucalyptus and Acacia into very local populations. There were no species that were distributed over the whole sampling area. The variation in leaf traits was larger between species than within species. Average nitrogen concentrations were 11.9 ± 1.05 mg g^?1 in Eucalyptus, and were 18.7 ± 4.1 mg g^?1 in Acacia. Even though the average nitrogen concentration was higher in Acacia than Eucalyptus, δ¹⁵N gave no clear indication for N₂ fixation in Acacia. In a multiple regression, latitude (as a surrogate for rainfall seasonality), mean rainfall, leaf nitrogen concentration, specific leaf area and nitrogen fixation were significant and explained 69 % of the variation of δ¹³C, but only 36 % of the variation of δ¹⁵N. Higher nitrogen and phosphorus concentration could give Acacia an advantage over Eucalyptus in arid regions of undefined rainfall seasonality.     

Natural 15N abundance of presumed N2-fixing and non-N2-fixing plants from selected ecosystems

Summary The ¹⁵N/¹⁴N ratios of plant and soil samples from Northern California ecosystems were determined by mass spectrometry. The ¹⁵N abundance of 176 plant foliar samples averaged 0.0008 atom % ¹⁵N excess relative to atmospheric N₂ and ranged from-0.0028 to 0.0064 atom % ¹⁵N excess relative to atmospheric N₂. Foliage from reported N₂-fixing species had significantly lower mean ¹⁵N abundance (relative to atmospheric N₂ and total soil N) and significantly higher N concentration (% N dry wt.) than did presumed non-N₂-fixing plants growing on the same sites. The mean difference between N₂-fixing species and other plants was 0.0007 atom % ¹⁵N. N₂-fixing species had lower ¹⁵N abundance than the other plants on most sites examined despite large differences between sites in vegetation, soil, and climate. The mean ¹⁵N abundance of N₂-fixing plants varied little between sites and was close to that of atmospheric N₂. The ¹⁵N abundance of presumed non-N₂-fixing species was highest at coastal sites and may reflect an input of marine spray N having relatively high ¹⁵N abundance. The ¹⁵N abundance of N₂-fixing species was not related to growth form but was for other plants. Annual herbaceous plants had highest ¹⁵N abundance followed in decreasing order by perennial herbs, shrubs, and trees. Several terrestrial ferns (Pteridaceae) had ¹⁵N abundances comparable to N₂-fixing legumes suggesting N₂-fixation by these ferns. On sites where the ¹⁵N abundance of soil N differs from that of the atmosphere, N₂-fixing plants can be identified by the natural ¹⁵N abundance of their foliage. This approach can be useful in detecting and perhaps measuring N₂-fixation on sites where direct recovery of nodules is not possible.     

Survival and colonization of inoculated bacteria in kallar grass rhizosphere and quantification of N2-fixation

Two experiments have been conducted, one in semi-solid Hoagland nutrient medium and the other in shallow pots containing saline soil. N₂-fixing bacteria belonging toAzospirillum, Azotobacter, Klebsiella andEnterobacter were inoculated separately on kallar grass grown in semi-solid nutrient medium. It was shown that inoculation affects root proliferation and also results in¹⁵N isotopic dilution. The % Ndfa ranged from 47–70 whereas no significant effect on the total nitrogen uptake was observed. The bacterial colonization of the root surface and the presence of enteric bacteria inside the root hair cells is reported. In a soil pot experiment, non-N₂-fixingPolypogon monspeliensis was used as a reference plant (control). A treatment receiving a high rate of nitrogen was also used as a non-N₂-fixing control.¹⁵N-labelled ammonium sulphate at 20 kg N ha^–1 and 90 kg N ha^–1 was used. The % Ndfa in the aerial parts of kallar grass was 12–15 whenP. monspeliensis was used as reference plant whereas 37–39% Ndfa was estimated when the treatment receiving high nitrogen fertilizer was used as a non-N₂-fixing control. These investigations revealed some problems of methodology which are discussed.     

Spatial variation of the stable nitrogen isotope ratio of woody plants along a topoedaphic gradient in a subtropical savanna

Variation in the stable N isotope ratio (δ¹⁵N) of plants and soils often reflects the influence of environment on the N cycle. We measured leaf δ¹⁵N and N concentration ([N]) on all individuals of Prosopis glandulosa (deciduous tree legume), Condalia hookeri (evergreen shrub), and Zanthoxylum fagara (evergreen shrub) present within a belt transect 308 m long × 12 m wide in a subtropical savanna ecosystem in southern Texas, USA in April and August 2005. Soil texture, gravimetric water content (GWC), total N and δ¹⁵N were also measured along the transect. At the landscape scale, leaf δ¹⁵N was negatively related to elevation for all the three species along this topoedaphic sequence. Changes in soil δ¹⁵N, total N, and GWC appeared to contribute to this spatial pattern of leaf δ¹⁵N. In lower portions of the landscape, greater soil N availability and GWC are associated with relatively high rates of both N mineralization and nitrification. Both soil δ¹⁵N and leaf [N] were positively correlated with leaf δ¹⁵N of non-N₂ fixing plants. Leaf δ¹⁵N of P. glandulosa, an N₂-fixing legume, did not correlate with leaf [N]; the δ¹⁵N of P. glandulosa’s leaves were closer to atmospheric N₂ and significantly lower than those of C. hookeri and Z. fagara. Additionally, at smaller spatial scales, a proximity index (which reflected the density and distance of surrounding P. glandulosa trees) was negatively correlated with leaf δ¹⁵N of C. hookeri and Z. fagara, indicating the N₂-fixing P. glandulosa may be important to the N nutrition of nearby non-N₂-fixing species. Our results indicate plant ¹⁵N natural abundance can reflect the extent of N retention and help us better understand N dynamics and plant-soil interactions at ecosystem and landscape scales.     

Comparative Genomic Analysis of N2-Fixing and Non-N2-Fixing Paenibacillus spp.: Organization,Evolution and Expression of the Nitrogen Fixation Genes

We provide here a comparative genome analysis of 31 strains within the genus Paenibacillus including 11 new genomic sequences of N₂-fixing strains. The heterogeneity of the 31 genomes (15 N₂-fixing and 16 non-N₂-fixing Paenibacillus strains) was reflected in the large size of the shell genome, which makes up approximately 65.2% of the genes in pan genome. Large numbers of transposable elements might be related to the heterogeneity. We discovered that a minimal and compact nif cluster comprising nine genes nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV encoding Mo-nitrogenase is conserved in the 15 N₂-fixing strains. The nif cluster is under control of a σ⁷⁰-depedent promoter and possesses a GlnR/TnrA-binding site in the promoter. Suf system encoding [Fe–S] cluster is highly conserved in N₂-fixing and non-N₂-fixing strains. Furthermore, we demonstrate that the nif cluster enabled Escherichia coli JM109 to fix nitrogen. Phylogeny of the concatenated NifHDK sequences indicates that Paenibacillus and Frankia are sister groups. Phylogeny of the concatenated 275 single-copy core genes suggests that the ancestral Paenibacillus did not fix nitrogen. The N₂-fixing Paenibacillus strains were generated by acquiring the nif cluster via horizontal gene transfer (HGT) from a source related to Frankia. During the history of evolution, the nif cluster was lost, producing some non-N₂-fixing strains, and vnf encoding V-nitrogenase or anf encoding Fe-nitrogenase was acquired, causing further diversification of some strains. In addition, some N₂-fixing strains have additional nif and nif-like genes which may result from gene duplications. The evolution of nitrogen fixation in Paenibacillus involves a mix of gain, loss, HGT and duplication of nif/anf/vnf genes. This study not only reveals the organization and distribution of nitrogen fixation genes in Paenibacillus, but also provides insight into the complex evolutionary history of nitrogen fixation.     

Australian mulga ecosystems –13C and 15N natural abundances of biota components and their ecophysiological significance

Samples of recently produced shoot material collected in winter/spring from common plant species of mulga vegetation in eastern and Western Australia were assayed for ¹³C and ¹⁵N natural abundance. ¹³C analyses showed only three of the 88 test species to exhibit C₄ metabolism and only one of seven succulent species to be in CAM mode. Non-succulent winter ephemeral C₃ species showed significantly lower mean δ¹³C values (– 28·0‰) than corresponding C₃-type herbaceous perennials, woody shrubs or trees (– 26·9, – 25·7 and – 26·2‰, respectively), suggesting lower water stress and poorer water use efficiency in carbon acquisition by the former than latter groups of taxa. Corresponding values for δ¹⁵N of the above growth and life forms lay within the range 7·5–15·5‰. δ¹⁵N of soil NH₄⁺ (mean 19·6‰) at a soft mulga site in Western Australia was considerably higher than that of NO₃^– (4·3‰). Shoot dry matter of Acacia spp. exhibited mean δ¹⁵N values (9·10 ± 0·6‰) identical to those of 37 companion non-N₂-fixing woody shrubs and trees (9·06 ± 0·5‰). These data, with no evidence of nodulation, suggested little or no input of fixed N₂ by the legumes in question. However, two acacias and two papilionoid legumes from a dune of wind-blown, heavily leached sand bordering a lake in mulga in Western Australia recorded δ¹⁵N values in the range 2·0–3·0‰ versus 6·4–10·7‰ for associated non-N₂-fixing taxa. These differences in δ¹⁵N, and prolific nodulation of the legumes, indicated symbiotic inputs of fixed N in this unusual situation. δ¹⁵N signals of lichens, termites, ants and grasshoppers from mulga of Western Australia provided evidence of N₂ fixation in certain termite colonies and by a cyanobacteria-containing species of lichen. Data are discussed in relation to earlier evidence of nitrophily and water availability constraints on nitrate utilization by mulga vegetation.     

Foliar δ15N is affected by foliar nitrogen uptake, soil nitrogen, and mycorrhizae along a nitrogen deposition gradient

Foliar nitrogen isotope (δ¹⁵N) composition patterns have been linked to soil N, mycorrhizal fractionation, and within-plant fractionations. However, few studies have examined the potential importance of the direct foliar uptake of gaseous reactive N on foliar δ¹⁵N. Using an experimental set-up in which the rate of mycorrhizal infection was reduced using a fungicide, we examined the influence of mycorrhizae on foliar δ¹⁵N in potted red maple (Acer rubrum) seedlings along a regional N deposition gradient in New York State. Mycorrhizal associations altered foliar δ¹⁵N values in red maple seedlings from 0.06 to 0.74 ‰ across sites. At the same sites, we explored the predictive roles of direct foliar N uptake, soil δ¹⁵N, and mycorrhizae on foliar δ¹⁵N in adult stands of A. rubrum, American beech (Fagus grandifolia), black birch (Betula lenta), and red oak (Quercus rubra). Multiple regression analysis indicated that ambient atmospheric nitrogen dioxide (NO₂) concentration explained 0, 69, 23, and 45 % of the variation in foliar δ¹⁵N in American beech, red maple, red oak, and black birch, respectively, after accounting for the influence of soil δ¹⁵N. There was no correlation between foliar δ¹³C and foliar %N with increasing atmospheric NO₂ concentration in most species. Our findings suggest that total canopy uptake, and likely direct foliar N uptake, of pollution-derived atmospheric N deposition may significantly impact foliar δ¹⁵N in several dominant species occurring in temperate forest ecosystems.     

Selecting matched root architecture in tree pairs to be used for assessing N2 fixation based on soil-15N-labelling

It is commonly assumed that soil-¹⁵N-labelling provides reliable estimates of N₂ fixation in trees by matching N₂-fixing and non-N₂-fixing tree pairs. As root system is a key parameter in determining suitability of the tree pairs, we compared root architecture of Acacia cyanophylla Lindl. and Casuarina glauca Sieber ex. Spreng. (two N₂-fixing trees) with Eucalyptus camaldulensis Dehn. and Ceratonia siliqua L. (two non-N₂-fixing trees) at 4-year-old in Mediterranean-semiarid zone. The rhizobium strain used appeared more motile than Frankia strain. A. cyanophylla and E. camaldulensis had extensive rooting area and volume of fine roots, and both species tended to develop marked horizontal rooting, compared to C. glauca and C. siliqua. Characteristics of fine- and horizontal-root components can be used in selecting matched root systems of N₂-fixing and reference-paired trees. Root architecture of C. glauca was more similar to C. siliqua, than to E. camaldulensis, and that of A. cyanophylla was more similar to E. camaldulensis than to C. siliqua. Accordingly, E. camaldulensis is an appropriate reference to estimate actual N₂ fixation by A. cyanophylla, and C. siliqua is an appropriate reference for C. glauca, when using soil-¹⁵N-labelling method in the prevailing site environment.     

An assessment of soil enrichment by actinorhizal N2 fixation using δ15N values in a chronosequence of deglaciation at Glacier Bay,Alaska

The extent of transfer of fixed N between N₂-fixing and non-N₂-fixing plant species is largely unknown in successional studies. In order to redress this deficiency at a locale intensively studied ecologically, leaf tissue samples were collected from actinorhizal N₂-fixing (Alnus, Shepherdia, and Dryas) and two non-N₂-fixing (Salix) woody species within research plots located along a chronosequence of deglaciated fjord in Glacier Bay National Park, Alaska. The tissue samples were analyzed for ¹⁵N content, and the resulting data analyzed for trends in plant tissue N. Among the non-N₂-fixing Salix species, ¹⁵N values increased from the most recently deglaciated sites to converge with the temporally more-stable values for the symbiotic N₂-fixing species on sites at about 40 years after deglaciation. The lower ¹⁵N values of sequestered N in plant tissues suggested that N derived from N₂-fixing plants accounts for the major portion of N in associated plants up to 40 years after deglaciation. The ¹⁵N isotopic data also suggested that Shepherdia canadensis depends least on soil N, D. drummondii the most, and A. viridis ssp. sinuata somewhere between those two species. The presence of a sere dominated by dense thickets of A. viridis ssp. sinuata at the convergence of ¹⁵N values for the N₂-fixing and non-N₂-fixing species indicated that this species is most responsible for accumulation of fixed N in soil at Glacier Bay. This paper is dedicated to the memory of Steven J. Kohls who died prior to publication of this research.     

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

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

Impacts of invading N2-fixing Acacia species on patterns of nutrient cycling in two Cape ecosystems: evidence from soil incubation studies and 15N natural abundance values

This study examines the impacts of woody, N₂-fixing invasive Acacia spp. on the patterns of nutrient cycling in two invaded ecosystems of differing nutrient status in the Cape floristic region. Patterns of soil nutrient mineralization were measured by a field incubation method while the significance of the fixation process in altering nutrient cycling was assessed by the ¹⁵N natural abundance technique. The results confirm earlier reports that invasion by woody shrubs results in organic matter and nutrient enrichment of surface soils of both ecosystems. However, patterns of nutrient availability (phosphorus and nitrogen) were not necessarily enhanced. In the more fertile strandveld both phosphorus and nitrogen (significant at P<0.10) showed trends towards enhanced annual mineralization rates upon invasion, while in the low nutrient fynbos system only phosphorus followed this trend. It is unclear whether this differential response is a consequence of plant- or soil-derived feedbacks on the decomposition processes in each system. The ¹⁵N values of the soils from the invaded sites of both ecosystems indicated a strong influence of the alien species on the soil nitrogen component. However, as with other studies of natural ecosystems, the contribution of nitrogen from fixation could not be readily quantified with the ¹⁵N natural abundance method because of problems in selecting suitable non-N₂-fixing reference plants. A technique of disrupting nodule structure and function, by fumigation with O₂, to obtain the ¹⁵N value of a non-N₂-fixing speciment of the study species was tried and found to overcome some of the problems associated with the lack of suitable reference plants. With this technique it was possible to detect the almost total dependence of A. saligna on N₂-fixation in the fynbos soils with their low nitrogen mineralization rates. In the strandveld ecosystem with much higher soil nitrogen release rates A. cyclops was only partly dependent on fixation (about half) for its nitrogen. The nutrient enrichment of both ecosystems and trends towards enhanced rates of nutrient mineralization could have profound implications on the long-term success of alien invader clearing operations and the restoration of the indigenous flora at these sites.     

Nitrogen and phosphorus retranslocation and N:P ratios of litterfall in three tropical plantations: luxurious N and efficient P use by Acacia mangium

Some tropical N₂-fixing trees exhibit specific characteristics for phosphorus (P) acquisition and utilisation that contrast with the large nitrogen (N) fluxes in their litterfall. To investigate differences in N and P cycling in N₂-fixing plantations, litterfall and fresh leaf quality of a N₂-fixing Acacia mangium plantation were compared with that of a non-N₂-fixing Swietenia macrophylla plantation and a coniferous Araucaria cunninghamii plantation. The N concentration in the A. mangium litterfall was higher than that in the litterfall of the two other species, whereas the P concentration in the A. mangium leaf litterfall was 0.16 mg g^–1, which was only 12–22% of that of the other species. The P concentration in the reproductive parts of A. mangium was markedly higher (16.1 mg g^–1) than those in the other fractions. The N:P ratio was higher in the leaf fall (81) compared to the fresh leaves (29) of A. mangium, in contrast to the N:P ratios in the leaf samples of the other two species. An analysis of a global litterfall dataset of tropical plantations indicated that N:P ratios in litterfall were significantly higher in N₂-fixers than in non-N₂-fixers, and those of A. mangium were high among species in the N₂-fixer group. These results indicated that A. mangium efficiently retranslocated P in contrast to very large N cycling, under field conditions. These differences may be related to other physiological characteristics of A. mangium.     

Thyme travels: 15N isoscapes of Thymus vulgaris L. invasion in lightly grazed pastoral communities

Alterations to ecosystem nitrogen (N) cycling by introduced plant species may increase the invasibility of habitat providing a positive feedback for the introduced species to become invasive. Spatial patterns of foliar and soil δ¹⁵N ratios reflect variation in rates and process of N‐cycling across invaded landscapes and provide insight into N‐source uptake and utilization strategies of invasive plant species. To evaluate invasion‐associated changes in soil and foliar δ¹⁵N at different scales: regional (among different sites), local (between north‐ and south‐facing aspect at the same site), and microsite (within populations in the same community), we measured foliar and soil δ¹⁵ N, animal faeces cover (as a proxy for grazing intensity) and N₂‐fixing species cover from inside to outside Thymus vulgaris L. (thyme)‐invaded lightly grazed pastoral communities in Central Otago, southern South Island, New Zealand. Mean thyme foliar δ¹⁵N were near‐zero across the invaded landscape, and did not change across the advancing edge of invasion or with aspect. There was no evidence that associations with N₂‐fixing species provide a potential N source. Soil δ¹⁵N was lower inside of thyme compared to at the edge or outside of thyme and was varied between aspects at some sites. Animal faeces cover as a proxy for grazing intensity explained only 23% of this observed variation of soil δ¹⁵N. Thyme invasion may result in lowered soil δ¹⁵N reflecting alterations to N dynamics. Associated invasion‐related impacts of animal grazing may also impact soil δ¹⁵N. Further studies are required to distinguish the underlying mechanism responsible for the observed patterns of foliar and soil δ¹⁵N values across thyme‐invaded Central Otago landscapes.     

Foliar and soil δ15N values reveal increased nitrogen partitioning among species in diverse grassland communities

Plant and soil nitrogen isotope ratios (δ¹⁵N) were studied in experimental grassland plots of varying species richness. We hypothesized that partitioning of different sources of soil nitrogen among four plant functional groups (legumes, grasses, small herbs, tall herbs) should increase with diversity. Four years after sowing, all soils were depleted in ¹⁵N in the top 5 cm whereas in non‐legume plots soils were enriched in ¹⁵N at 5–25 cm depth. Decreasing foliar δ¹⁵N and Δδ¹⁵N (= foliar δ¹⁵N ? soil δ¹⁵N) values in legumes indicated increasing symbiotic N₂ fixation with increasing diversity. In grasses, foliar Δδ¹⁵N also decreased with increasing diversity suggesting enhanced uptake of N depleted in ¹⁵N. Foliar Δδ¹⁵N values of small and tall herbs were unaffected by diversity. Foliar Δδ¹⁵N values of grasses were also reduced in plots containing legumes, indicating direct use of legume‐derived N depleted in ¹⁵N. Increased foliar N concentrations of tall and small herbs in plots containing legumes without reduced foliar δ¹⁵N indicated that these species obtained additional mineral soil N that was not consumed by legumes. These functional group and species specific shifts in the uptake of different N sources with increasing diversity indicate complementary resource use in diverse communities.     

Nitrogen Transfer Within and Between Plants Through Common Mycorrhizal Networks (CMNs)

Mycorrhizae play a critical role in nutrient capture from soils. Arbuscular mycorrhizae (AM) and ectomycorrhizae (EM) are the most important mycorrhizae in agricultural and natural ecosystems. AM and EM fungi use inorganic NH₄ ⁺ and NO₃ ^?, and most EM fungi are capable of using organic nitrogen. The heavier stable isotope ¹⁵N is discriminated against during biogeochemical and biochemical processes. Differences in ¹⁵N (atom%) or δ¹⁵N (‰) provide nitrogen movement information in an experimental system. A range of 20 to 50% of one-way N-transfer has been observed from legumes to nonlegumes. Mycorrhizal fungal mycelia can extend from one plant's roots to another plant's roots to form common mycorrhizal networks (CMNs). Individual species, genera, even families of plants can be interconnected by CMNs. They are capable of facilitating nutrient uptake and flux. Nutrients such as carbon, nitrogen and phosphorus and other elements may then move via either AM or EM networks from plant to plant. Both ¹⁵N labeling and ¹⁵N natural abundance techniques have been employed to trace N movement between plants interconnected by AM or EM networks. Fine mesh (25～45 μm) has been used to separate root systems and allow only hyphal penetration and linkages but no root contact between plants. In many studies, nitrogen from N₂-fixing mycorrhizal plants transferred to non-N₂–fixing mycorrhizal plants (one-way N-transfer). In a few studies, N is also transferred from non-N₂–fixing mycorrhizal plants to N₂-fixing mycorrhizal plants (two-way N-transfer). There is controversy about whether N-transfer is direct through CMNs, or indirect through the soil. The lack of convincing data underlines the need for creative, careful experimental manipulations. Nitrogen is crucial to productivity in most terrestrial ecosystems, and there are potential benefits of management in soil-plant systems to enhance N-transfer. Thus, two-way N-transfer warrants further investigation with many species and under field conditions.     

Soil Organic Carbon is Increased in Mixed-Species Plantations of Eucalyptus and Nitrogen-Fixing Acacia

Soil organic carbon (soil C) sequestration in forests is often higher under nitrogen (N₂)-fixing than under non-N₂-fixing tree species. Here, we examined whether soil C could be increased using mixed-species plantations compared to monocultures, which are less productive aboveground than mixtures. In addition, we compared soil C sequestration under N₂-fixing trees with non-N₂-fixing trees that received N fertilizer. Monocultures of Eucalyptus globulus (E) and the N₂-fixing Acacia mearnsii (A) and mixtures of these species were planted in a replacement series: 100%E, 75%E + 25%A, 50%E + 50%A, 25%E + 75%A and 100%A. Soil samples were also collected from fertilized monoculture treatments (100%EFer) of E. globulus (250 kg N ha^?1). Total organic C, N and phosphorus were determined at age 8 years at two soil depths (0–10 cm and 10–30 cm) and three density fractions of soil organic matter (SOM) were quantified for 0–5 cm depth. Soil C was highest in the 50%E + 50%A mixed stand and was highly correlated with aboveground biomass, not to the percentage of A. mearnsii in mixtures. This was largely due to soil C at 10–30 cm because there were no treatment effects on soil C at 0–10 cm. All density fractions of SOM at 0–5 cm increased with the percentage of A. mearnsii. In E. globulus monocultures, N fertilization did not increase soil C when compared with unfertilized stands. These results indicate that the inclusion of N₂-fixing trees into eucalypt plantations may increase soil C stocks through increased productivity.     

N-fertilization does not alleviate grass competition induced reduction of growth of African savanna species

Background and aims

Below-ground grass competition limits woody establishment in savannas. N₂-fixing legumes may, however, have a nutritional advantage over broad-leaved species. We hypothesised that broad-leaved non-legume savanna thicket species would be more severely constrained by grass competition for N and consequently respond more to N-fertilization than the legume, Acacia karroo.

Methods

A. karroo and five non-legume thicket species (Maytenus senegalensis, M. heterophylla, Euclea divinorum, Ziziphus mucronata, Schotia brachypetala) were grown together in an irrigated competition experiment with clipped-, unclipped-grass and without grass with/without N-fertilizer. The biomass, foliar nutrient, δ¹³C and δ¹⁵N of grasses and woody species were determined.

Results

Growth of both A. karroo and the non-legume species was equally sensitive (c. 90 % reduction) to both clipped- and unclipped-grass competition, regardless of N-fertilization. With grass competition, however, foliar [N] increased and δ¹⁵N decreased in response to N-fertilization. Grass biomass accumulation was also unchanged by fertilisation, despite increases in foliar [N] and decreases in δ¹⁵N.

Conclusions

The N₂-fixation capacity of A. karroo provided no growth advantage over non-legumes. The lack of responsiveness of biomass accumulation by both the woody species and the grasses to N-fertilization, despite evidence that plants accessed the N-fertilizer, indicates limitation by other nutrients.     

Effects of elevated CO<Subscript>2</Subscript> and nitrogen on wood structure related to water transport in seedlings of two deciduous broad-leaved tree species

Wood structure might be altered through the physiological responses to atmospheric carbon dioxide concentration ([CO₂]) and nitrogen (N) deposition. We investigated growth, water relations and wood structure of 1-year-old seedlings of two deciduous broad-leaved tree species, Quercus mongolica (oak, a ring-porous species) and Alnus hirsuta (alder, a diffuse-porous species and N₂–fixer), grown under a factorial combination of two levels of [CO₂] (36 and 72 Pa) and nitrogen supply (N; low and high) for 141 days in phytotron chambers. In oak, there was no significant effect of [CO₂] on wood structure, although elevated [CO₂] tended to decrease stomatal conductance (g _s) and increased water use efficiency regardless of the N treatment. However, high N supply increased root biomass and induced wider earlywood and larger vessels in the secondary xylem in stems, leading to increased hydraulic conductance. In alder, there was significant interactive effect of [CO₂] and N on vessel density, and high N supply increased the mean vessel area. Our results suggest that wood structures related to water transport were not markedly altered, although elevated [CO₂] induced changes in physiological parameters such as g _s and biomass allocation, and that N fertilization had more pronounced effects on non-N₂-fixing oak than on N₂-fixing alder.     

Symbiotic nitrogen fixation in eight Acacia senegal provenances in dryland clays of the Blue Nile Sudan estimated by the 15N natural abundance method

The symbiotic biological N₂fixation by Acacia senegal was estimated using the ¹⁵N natural abundance (δ ¹⁵N) procedure on eight provenances collected from different environments and soil types grown in a clay soil in the Blue Nile region, Sudan. Balanites aegyptiaca (a non-legume) was used as a non-N₂-fixing reference plant to allow ¹⁵N-based estimates of the proportion of the Acacia N derived from atmospheric N₂ (N_dfa) to be calculated. Results show variation in leaf δ ¹⁵N between A. senegal and the reference plant and among years. The relative δ ¹⁵N values (‰) were higher in B. aegyptiaca than in the N₂-fixing acacia provenances. Provenances originally collected from clay soils fixed little N in the first year, but the amount fixed increased as the trees aged. All provenances showed a decrease in δ ¹⁵N with age. The N_dfa varied between 24% (Mazmoom provenance) and 61% (Rahad provenance) 4 years after planting. There was no significant difference in δ ¹⁵N between provenance groups based on soil type or rainfall at original growing site. The amount of N_dfa increased significantly with age in all provenances. The above-ground contribution of fixed N to foliage growth in a 4-year-old A. senegal was highest in the Rahad sand–soil provenance (46.7 kg N ha⁻¹) and lowest in the Mazmoom clay-soil provenance (28.7 kg N ha⁻¹). Our study represents the first use of the δ ¹⁵N method for estimating the N input by A. senegal to the clay plain soils of the gum belt in the Sudan.