Nitrogen fertilisation reduces grass-induced N2 fixation of tree seedlings from semi-arid savannas

Aims

Coexistence of trees and grasses in nutrient-poor arid savannas may result in competition for soil N. While grasses may be more effective than woody plants in acquiring N from the soil, some leguminous woody species rely on N₂ fixation. We assessed the role of N₂ fixation in the N-budget of Acacia mellifera seedlings by varying N supply and grass competition.

Methods

The contribution of N₂ fixation to the N-budget of Acacia mellifera seedlings with varying N supply and grass competition was determined by measuring growth, nutrient concentrations, and ¹⁵N values.

Results

Tree seedlings were 4-fold taller and had 20-fold more biomass in the absence of grass. Tree foliar δ¹⁵N was lower with (?0.25?±?0.2‰, n?=?9) than without grasses (5.2?±?0.1‰, n?=?64). The contribution of N₂-fixation to the N budget decreased with increasing N supply. Greater reliance on N₂-fixation by trees in the presence of grasses did not result in greater biomass accumulation or tissue [N] relative to tree seedlings grown without grass competition. Tree seedlings competing with grass had significantly more negative δ¹³C (?29.5?±?0.6‰) than seedlings without grass competition (?28.8‰?±?0.5‰).

Conclusions

Induction of N₂-fixation by grass may have resulted from competition for nutrients. N₂-fixation enables tree seedlings to compensate for limited soil N and survive grass competition at a critical and vulnerable developmental stage of germination and establishment.     

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.     

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.     

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.     

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.     

Inoculation of forage grasses with N2-fixing Enterobacteriaceae

Summary Previous investigations indicated some forage grass roots in Texas are heavily colonized with N₂-fixing bacteria. The most numerous N₂-fixing bacteria were in the genera Klebsiella and Enterobacter. In the present investigation inoculation experiments were conducted using 18 isolates of these bacteria to determine if a N₂-fixing association could be established between the bacteria and the grassesCynodon dactylon andPanicum coloratum. Plants were grown in soil for approximately 5 months in a greenhouse and were measured periodically for dry matter, nitrogen accumulation, and acetylene reduction activity. Results of the investigation indicated that 25% of the plant-soil systems were active in acetylene reduction and the activity was high enough to indicate agronomically significant quantities of N₂ were being fixed (>8kg N ha⁻¹). However, plant systems extrapolated to fix>8 kg N ha⁻¹ contained less nitrogen and accumulated less dry matter than plants less active in acetylene reduction. Inocula could not be re-isolated from healthy grass roots indicating that the N₂-fixing activity may have not have been closely assiciated with plant roots. Future research is needed to determine factors limiting colonization of grass roots.     

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.     

Sugar beet and barley yields in relation to inoculation with N2-fixing and phosphate solubilizing bacteria

Recently, there has been a resurgence of interest in bioorganic fertilizers as part of sustainable agricultural practices to alleviate drawbacks of intensive farming practices. N₂-fixing and P-solubilizing bacteria are important in plant nutrition increasing N and P uptake by the plants, and playing a significant role as plant growth-promoting rhizobacteria in the biofertilization of crops. A study was conducted in order to investigate the effects of two N₂-fixing (OSU-140 and OSU-142) and a strain of P-solubilizing bacteria (M-13) in single, dual and three strains combinations on sugar beet and barley yields under field conditions in 2001 and 2002. The treatments included: (1) Control (no inoculation and fertilizer), (2) Bacillus OSU-140, (3) Bacillus OSU-142, (4) Bacillus M-13, (5) OSU-140 + OSU-142, (6) OSU-140 + M-13, (7) OSU-142 + M-13, (8) OSU-140 + OSU-142 + M-13, (9) N, (10) NP. N and NP plots were fertilized with 120 kg N ha^–1 and 120 kg N ha^–1 + 90 kg P ha^- for sugar beet and 80 kg N ha^–1 and 80 kg N ha^–1 + 60 kg P ha^–1 for barley. The experiments were conducted in a randomized block design with five replicates. All inoculations and fertilizer applications significantly increased leaf, root and sugar yield of sugar beet and grain and biomass yields of barley over the control. Single inoculations with N₂-fixing bacteria increased sugar beet root and barley yields by 5.6–11.0% depending on the species while P-solubilizing bacteria alone gave yield increases by 5.5–7.5% compared to control. Dual inoculation and mixture of three bacteria gave increases by 7.7–12.7% over control as compared with 20.7–25.9% yield increases by NP application. Mixture of all three strains, dual inoculation of N₂-fixing OSU-142 and P-solubilizing M-13, and/or dual inoculation N₂-fixing bacteria significantly increased root and sugar yields of sugar beet, compared with single inoculations with OSU-140 or M-13. Dual inoculation of N₂-fixing Bacillus OSU-140 and OSU-142, and/or mixed inoculations with three bacteria significantly increased grain yield of barley compared with single inoculations of OSU-142 and M-13. In contrast with other combinations, dual inoculation of N₂-fixing OSU-140 and P-solubilizing M-13 did not always significantly increase leaf, root and sugar yield of sugar beet, grain and biomass yield of barley compared to single applications both with N₂-fixing bacteria. The beneficial effects of the bacteria on plant growth varied significantly depending on environmental conditions, bacterial strains, and plant and soil conditions.     

Analysis of environmental drivers influencing interspecific variations and associations among bloom-forming cyanobacteria in large,shallow eutrophic lakes

Non-diazotrophic Microcystis and filamentous N₂-fixing Aphanizomenon and Dolichospermum (formerly Anabaena) co-occur or successively dominate freshwaters globally. Previous studies indicate that dual nitrogen (N) and phosphorus (P) reduction is needed to control cyanobacterial blooms; however, N limitation may cause replacement of non-N₂-fixing by N₂-fixing taxa. To evaluate potentially counterproductive scenarios, the effects of temperature, nutrients, and zooplankton on the spatio-temporal variations of cyanobacteria were investigated in three large, shallow eutrophic lakes in China. The results illustrate that the community composition of cyanobacteria is primarily driven by physical factors and the zooplankton community, and their interactions. Niche differentiation between Microcystis and two N₂-fixing taxa in Lake Taihu and Lake Chaohu was observed, whereas small temperature fluctuations in Lake Dianchi supported co-dominance. Through structural equation modelling, predictor variables were aggregated into ‘composites’ representing their combined effects on species-specific biomass. The model results showed that Microcystis biomass was affected by water temperature and P concentrations across the studied lakes. The biomass of two filamentous taxa, by contrast, exhibited lake-specific responses. Understanding of driving forces of the succession and competition among bloom-forming cyanobacteria will help to guide lake restoration in the context of climate warming and N:P stoichiometry imbalances.     

Arbuscular mycorrhizal fungi affect seedling recruitment: a potential mechanism by which N deposition favors the dominance of grasses over forbs

Background and aims

Nitrogen (N) deposition usually alters plant community structure and reduces plant biodiversity in grasslands. Seedling recruitment is essential for maintaining species richness and determines plant community composition. Arbuscular mycorrhizal fungi (AMF) are widespread symbiotic fungi and could facilitate seedling establishment. Here we conducted an experiment to address whether the influence of AMF on seedling recruitment depends on N addition and plant species.

Methods

Leymus chinensis were cultivated for 5 months in the microcosms that were inoculated with or without AMF at five N addition rates. Seeds of three main species (two C₃ grasses and one non-N₂-fixing forb) of the Eurasian steppe were sown to the 5-month-old microcosms. Seedling establishment was estimated by shoot biomass, N and P contents 7 weeks after seedling germination.

Results

AMF promoted seedlings recruitment of two C₃ grasses at addition rates above 0.5 g N m^?2. In contrast, seedling recruitment of the non-N₂-fixing forb was increased by AMF at addition rates below 0.5 g N m^?2 but was decreased above 2.5 g N m^?2.

Conclusions

These results partly explain why N addition favored the dominance of grasses over forbs in perennial grassland communities. Our study indicates that AMF have the potential to influence plant community composition by mediating revegetation in the face of N deposition.     

N2O and CH4 Emissions,and NO3 − Leaching on a Crop-Yield Basis from a Subtropical Rain-fed Wheat–Maize Rotation in Response to Different Types of Nitrogen Fertilizer

Guaranteeing high crop yields while reducing environmental impacts of nitrogen fertilizer use due to associated losses of N₂O emissions and nitrate (NO₃ ^?) leaching is a key challenge in the context of sustainable intensification of crop production. However, few field data sets are available that explore the effect of different forms of N management on yields as well as on N losses in the form of N₂O or NO₃ ^?. Here we report on a large-scale field lysimeter (8 × 4 m²) experiment, which was designed to determine soil CH₄ and N₂O emissions, NO₃ ^? leaching losses and crop yields from a subtropical rain-fed wheat–maize rotation in the Sichuan Basin, one of the most intensively used agricultural regions in China. One control and three different fertilizer treatments with the same total rate of N application (280 kg N ha^?1 y^?1) were included: NF: control (no fertilizer); NPK: synthetic N fertilizer; OMNPK: synthetic N fertilizer plus pig manure; RSDNPK: synthetic N fertilizer plus crop residues. As compared to the standard NPK treatment, annual NO₃ ^? leaching losses for OMNPK and RSDNPK treatments were decreased by 36 and 22%, respectively (P < 0.05). Similarly, crop yield-scaled NO₃ ^? leaching for NPK treatment was higher than those for either OMNPK or RSDNPK treatments (P < 0.05). Direct N₂O emissions for RSDNPK treatment were decreased as compared with NPK and OMNPK treatments (P < 0.05). Furthermore, the yield-scaled GWP (global warming potential) was lower for the treatments where either pig manure or crop residues were incorporated as compared to the standard NPK treatment (P < 0.05). Our study indicates that it is possible to reduce the negative environmental impact of NO₃ ^? leaching and N₂O emissions without compromising crop productivity. Yield-scaled NO₃ ^? leaching, similar to the yield-scaled GWP, represents another valuable-integrated metric to address the dual goals of reducing nitrogen pollution and maintaining crop grain yield for a given agricultural system.     

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.     

Soil nitrous oxide emissions from a typical semiarid temperate steppe in inner Mongolia: effects of mineral nitrogen fertilizer levels and forms

Nitrous oxide (N₂O) emissions can be significantly affected by the amounts and forms of nitrogen (N) available in soils, but the effect is highly dependent on local climate and soil conditions in specific ecosystem. To improve our understanding of the response of N₂O emissions to different N sources of fertilizer in a typical semiarid temperate steppe in Inner Mongolia, a 2-year field experiment was conducted to investigate the effects of high, medium and low N fertilizer levels (HN: 200 kg N?ha^-1y^-1, MN: 100 kg N ha^-1y^-1, and LN: 50 kg N ha^-1y^-1) respectively and N fertilizer forms (CAN: calcium ammonium nitrate, AS: ammonium sulphate and NS: sodium nitrate) on N₂O emissions using static closed chamber method. Our data showed that peak N₂O fluxes induced by N treatments were concentrated in short periods (2 to 3 weeks) after fertilization in summer and in soil thawing periods in early spring; there were similarly low N₂O fluxes from all treatments in the remaining seasons of the year. The three N levels increased annual N₂O emissions significantly (P?<?0.05) in the order of MN > HN > LN compared with the CK (control) treatment in year 1; in year 2, the elevation of annual N₂O emissions was significant (P?<?0.05) by HN and MN treatments but was insignificant by LN treatments (P?>?0.05). The three N forms also had strong effects on N₂O emissions. Significantly (P?<?0.05) higher annual N₂O emissions were observed in the soils of CAN and AS fertilizer treatments than in the soils of NS fertilizer treatments in both measured years, but the difference between CAN and AS was not significant (P?>?0.05). Annual N₂O emission factors (EF) ranged from 0.060 to 0.298% for different N fertilizer treatments in the two observed years, with an overall EF value of 0.125%. The EF values were by far less than the mean default EF proposed by the Intergovernmental Panel on Climate Change (IPCC).     

Topographical influences on foliar nitrogen concentration and stable isotope composition in a Mediterranean-climate catchment

Nitrogen (N) oligotrophication is increasing globally across terrestrial ecosystems and manifested in decreasing nitrogen concentration ([N]) and changes in the stable nitrogen isotope composition (δ¹⁵N) of foliage. Heterogeneity in plant nitrogen sources makes it challenging to detect the effects of N oligotrophication even at a small catchment scale with complex topography. Understanding the spatial and temporal variation of foliar δ¹⁵N and [N] at such a scale is required to develop useful ecological indicators and monitoring methods to support catchment management with a potential N oligotrophication problem. This study examined spatial and high-resolution temporal variation of foliar δ¹⁵N and [N] and their influencing factors in ten trees grouped by Eucalyptus and Acacia in a native forest vegetation catchment. Over 16 sampling campaigns within a 12-month period, foliar δ¹⁵N and [N] increased in Eucalyptus but were constant in the N₂-fixing Acacia. The higher foliar [N] and δ¹⁵N in Acacia reflected its N₂-fixation ability. Topographic flow accumulation area (NDVI) explained 46% (77%) of spatial variation in dry-season Eucalyptus foliar δ¹⁵N ([N]). For Eucalyptus, foliar δ¹⁵N was higher at the downslope than the upslope locations, but no hillslope location differences were observed for foliar [N]. These results suggest that in the non-N₂-fixing Eucalyptus, seasonal water stress related nitrogen availability may be reflected in foliar δ¹⁵N rather than foliar [N]. As such, foliar δ¹⁵N of non-N₂-fixing plants potentially is a more sensitive indicator of seasonal or topographical N availability than foliar [N].     

Management of Warm- and Cool-Season Grasses for Biomass on Marginal Lands: II. Composition and Nutrient Balance

Organic fertilizers can improve soil health while providing nutrients for perennial grass growth for bioenergy feedstock, particularly under marginal soil conditions. The impact of organic fertilizer application on perennial grass composition needs clarification. Our objective was to evaluate feedstock composition, and N, P, and K dynamics of switchgrass (Panicum virgatum L.), tall fescue [Lolium arundinaceum (Schreb.)], and reed canarygrass (Phalaris arundinacea L.) provided with either inorganic or organic fertilizer sources. Grasses were established on a sandy soil and a clay soil at the Cornell University Willsboro Research Farm in Willsboro, NY. The experiment was a split-split plot randomization of a randomized block design with six replicates. Sites were whole plots, grass species were subplots, and fertility treatments were sub-subplots. Six treatments were (1) 168 kg ha^?1 of N fertilizer for cool-season grasses; 84 kg ha^?1 for switchgrass, (2) 56 kg ha^?1 of 0-46-0 P fertilizer plus N (#1), (3) 112 kg ha^?1 of 0-0-60 K fertilizer plus N (#1), (4) 89.6 Mg dairy manure ha^?1, (5) 44.8 Mg dairy manure compost ha^?1, and (6) a control without fertilizer. Organic fertilizers produced a net positive P and K balance, while other treatments had negative balances. Organic fertilizer treatments resulted in lower lignin and gross energy values, and higher total ash and Cl, compared to inorganic fertilizer treatments. Switchgrass biomass had higher fiber and gross energy, lower total ash, and much lower Cl content under organic fertilizer applications than cool-season grasses, making switchgrass a more desirable feedstock regardless of conversion process.     

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.     

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.     

Greenhouse Gas Emissions, Nitrate Leaching, and Biomass Yields from Production of Miscanthus × giganteus in Illinois, USA

Understanding the effects of nitrogen (N) fertilization on Miscanthus × giganteus greenhouse gas emissions, nitrate leaching, and biomass production is an important consideration when using this grass as a biomass feedstock. The objective of this study was to determine the effect of three N fertilization rates (0, 60, and 120?kg?N?ha^?1 using urea as the N source) on nitrous oxide (N₂O) and carbon dioxide (CO₂) emissions, nitrogen leaching, and the biomass yields and N content of M. × giganteus planted in July 2008, and evaluated from 2009 through early 2011 in Urbana, Illinois, USA. While there was no biomass yield response to N fertilization rates in 2009 and 2010, the amount of N in the harvested biomass in 2010 was significantly greater at the 60 and 120?kg?N?ha^?1?N rates. There was no significant CO₂ emission response to N rates in 2009 or 2010. Similarly, N fertilization did not increase cumulative N₂O emissions in 2009, but cumulative N₂O emissions did increase in 2010 with N fertilization. During 2009, nitrate (NO ₃ ^? ) leaching at the 50-cm soil depth was not related to fertilization rate, but there was a significant increase in NO ₃ ^? leaching between the 0 and 120?kg?N?ha^?1 treatments in 2010 (8.9 and 28.9?kg?NO₃?CN?ha^?1?year^?1, respectively). Overall, N fertilization of M. × giganteus led to N₂O releases, increased fluxes of inorganic N (primarily NO ₃ ^? ) through the soil profile; and increased harvested N without a significant increase in biomass production.     

Interactions between perennial ryegrass ( Lolium perenne L.) and white clover (Trifolium repens L.) under contrasting nitrogen availability: productivity,seasonal patterns of species composition,N2 fixation,N transfer and N recovery

Nitrogen (N) fertiliser and clover cultivar choice affect competition and productivity in grass-clover mixtures. Pure stands and mixtures of perennial ryegrass and white clover cultivars with contrasting growth habits were examined. The aim of this work was to study the effect of repetitive nitrogen (N) application and cultivar combination on competition and productivity, N yield in the harvested herbage, N₂ fixation in mixtures and pure stands, and transfer of N from clover to the companion grass. Large-leaved white clover cultivar Alice and small-leaved cv. Gwenda and perennial ryegrass cvs. Barlet (erect) and Heraut (prostrate) were sown in pure stands and as four binary grass-clover mixtures on a sandy soil in 1995. In the mixtures, two levels of N fertiliser were applied: 0 (-N) and 150 and 180 kg ha^-1 y^-1 N (+N) in 1996 and 1997, respectively, while the grass monocultures received three N levels (0, 140/180 and 280/360 kg ha^-1) in 1996 and 1997, respectively. No N was applied to pure clover. The plots were cut five times during 1996 and six times during 1997. Fertiliser N was applied in early spring and after every harvest. The treatments were continued until the summer of 1999. In pure grass, the applied N was effectively recovered. In mixtures, N application affected competition by enhancing grass growth and the overall effect of N application was 17 kg DM per kg N applied in 1996. However, there was no yield response to N fertilizer in 1997, because this was compensated for by a higher clover production in unfertilised mixtures. In 1997, -N mixtures yielded more N than +N mixtures, owing to the higher clover content and N₂ fixation. Large-leaved clover cv. Alice was better able to withstand the negative effect of repetitive N application on clover production in mixtures and increased its proportion during the growing season of the second harvest year. In 1997, mixtures with Alice yielded more N than mixtures with Gwenda, but in pure clover swards, there was no cultivar effect on N yield. Also, during the autumn of 1998 and the spring of 1999, the clover content was highest in mixtures with Alice. Harvested N and apparent N₂ fixation were almost twice as high in 1997 as in 1996. N yield and apparent N₂ fixation were higher in pure clover than in mixtures. In mixtures, the apparent N₂ fixation in 1996 was 142 kg N ha^-1, irrespective of cultivar or N treatment. In 1997, it was on average 337 kg N ha^-1, and higher in -N mixtures and in mixtures with Alice. For each tonne of clover DM in the harvested herbage, 65 and 57 kg N was harvested in 1996 and 1997 in -N mixtures, respectively. The apparent transfer of clover-derived N to grass was on average 29 and 70 kg N ha^-1 yr^-1 in 1996 and 1997, respectively. It was highest in +N mixtures and highest in mixtures with Gwenda in 1997. In contrast to clover, the grass cultivars were very similar in their productivity and seasonal patterns, despite their contrasting growth habits. Seasonal trends in N yield, N transfer and N recovery are discussed in relation to fertilizer application regimes and variation in production patterns in mixtures and pure stands. This revised version was published online in June 2006 with corrections to the Cover Date.