Biological nitrogen fixation in mixed legume/grass pastures

Biological nitrogen fixation (BNF) in mixed legume/grass pastures is reviewed along with the importance of transfer of fixed nitrogen (N) to associated grasses. Estimates of BNF depend on the method of measurement and some of the advantages and limitations of the main methods are outlined. The amounts of N fixed from atmospheric N₂ in legume/grass pastures throughout the world is summarised and range from 13 to 682 kg N ha^-1 yr^-1. the corresponding range for grazed pastures, which have been assessed for white clover pastures only, is 55 to 296 kg N ha^-1 yr^-1.Biological nitrogen fixation by legumes in mixed pastures is influenced by three primary factors; legume persistence and production, soil N status, and competition with the associated grass(es). These factors and the interactions between them are discussed. Legume persistence, production and BNF is also influenced by many factors and this review centres on the important effects of soil moisture status, soil acidity, nutrition, and pests and disease.Soil N status interacts directly with BNF in the short and long term. In the short-term, increases in soil inorganic N occurs during dry conditions and where N fertiliser is used, and these will reduce BNF. In the long-term, BNF leads to accumulation of soil N, grass dominance, and reduced BNF. However, cyclical patterns of legume and grass dominance can occur due, at least in part, to temporal changes in plant-available N levels in soil. Thus, there is a dynamic relationship between legumes and grasses whereby uptake of soil N by grass reduces the inhibitory effect of soil N on BNF and competition by grasses reduces legume production and BNF. Factors affecting the competition between legumes and grasses are considered including grass species, grazing animals, and grazing or cutting management.Some fixed N is transferred from legumes to associated grasses. The amount of N transferred below-ground, predominantly through decomposition of legume roots and nodules, has been estimated at 3 to 102 kg N ha^-1 yr^-1 or 2 to 26% of BNF. In grazed pasture, N is also transferred above-ground via return in animal excreta and this can be of a similar magnitude to below-ground transfer.Increased BNF in mixed legume/grass pastures is being obtained through selection or breeding of legumes for increased productivity and/or to minimise effects of nutrient limitations, low soil moisture, soil acidity, and pests and disease. Ultimately, this will reduce the need to modify the pasture environment and increase the role of legumes in low-input, sustainable agriculture.     

Factors regulating the contributions of fixed nitrogen by pasture and crop legumes to different farming systems of eastern Australia

On-farm and experimental measures of the proportion (%Ndfa) and amounts of N₂ fixed were undertaken for 158 pastures either based on annual legume species (annual medics, clovers or vetch), or lucerne (alfalfa), and 170 winter pulse crops (chickpea, faba bean, field pea, lentil, lupin) over a 1200 km north-south transect of eastern Australia. The average annual amounts of N₂ fixed ranged from 30 to 160 kg shoot N fixed ha^–1 yr^–1 for annual pasture species, 37–128 kg N ha^–1 yr^–1 for lucerne, and 14 to 160 kg N ha^–1 yr^–1 by pulses. These data have provided new insights into differences in factors controlling N₂ fixation in the main agricultural systems. Mean levels of %Ndfa were uniformly high (65–94%) for legumes growing at different locations under dryland (rainfed) conditions in the winter-dominant rainfall areas of the cereal-livestock belt of Victoria and southern New South Wales, and under irrigation in the main cotton-growing areas of northern New South Wales. Consequently N₂ fixation was primarily regulated by biomass production in these areas and both pasture and crop legumes fixed between 20 and 25 kg shoot N for every tonne of shoot dry matter (DM) produced. Nitrogen fixation by legumes in the dryland systems of the summer-dominant rainfall regions of central and northern New South Wales on the other hand was greatly influenced by large variations in %Ndfa (0–81%) caused by yearly fluctuations in growing season (April–October) rainfall and common farmer practice which resulted in a build up of soil mineral-N prior to sowing. The net result was a lower average reliance of legumes upon N₂ fixation for growth (19–74%) and more variable relationships between N₂ fixation and DM accumulation (9–16 kg shoot N fixed/t legume DM). Although pulses often fixed more N than pastures, legume-dominant pastures provided greater net inputs of fixed N, since a much larger fraction of the total plant N was removed when pulses were harvested for grain than was estimated to be removed or lost from grazed pastures. Conclusions about the relative size of the contributions of fixed N to the N-economies of the different farming systems depended upon the inclusion or omission of an estimate of fixed N associated with the nodulated roots. The net amounts of fixed N remaining after each year of either legume-based pasture or pulse crop were calculated to be sufficient to balance the N removed by at least one subsequent non-legume crop only when below-ground N components were included. This has important implications for the interpretation of the results of previous N₂ fixation studies undertaken in Australia and elsewhere in the world, which have either ignored or underestimated the N present in the nodulated root when evaluating the contributions of fixed N to rotations.     

Key processes of the nitrogen cycle in an irrigated and a non-irrigated grazed pasture

The paper presents integrated measurements of N fixation, net mineralisation, pasture yield and change in soil mineral N over a 12 month period for dairy pastures on a sandy loam soil in the South East of South Australia. The two adjacent pastures studied were an irrigated perennial white clover-ryegrass and an annual non-irrigated subterranean clover with mixed annual grasses. This produced the most comprehensive mineral N balance reported for grazed pastures, to the authors' knowledge, allowing calculation of gaseous and leaching losses of N (210 kg ha^–1 in the irrigated and paddock and 81 kg ha^–1 in the non irrigated paddock) primarily from urine patches. In both paddocks these losses were about three times the N yield in milk (61 and 28 kg N ha^–1 respectively) and were replenished by biological N fixation (294 and 100 kg N ha^–1). However, mineralisation of soil organic N, excretal N and pasture residues (687 and 438 kg N ha^–1) was the major source of mineral N for cycling and losses. The results demonstrate the enormous impact of pasture management on N fluxes and reinforce the importance of livestock urine on the magnitude of N fluxes including gaseous and leaching losses.     

Role of legumes in providing N for sustainable tropical pasture systems

Forage legumes have long been lauded for their ability to fix atmospheric nitrogen and contribute to the sustainability of agricultural production systems. However despite the benefits they bring in terms of increased herbage and animal production they are not widely used in temperate or tropical regions. In this review the amounts of biological nitrogen fixation (BNF) needed to sustain the soil-plant-animal system are discussed and related to the amounts fixed in tropical pastures. The data suggest that tropical forage legumes have the capacity to meet the requirements to balance the N cycle of grazed pastures. The actual amounts required will depend on the rate of pasture utilization and the efficiency of recycling via litter, excreta and internal remobilization. The efficiency of nitrogen fixation (% of legume N derived from fixation) is usually high in tropical pastures (>80%) and is unlikely to be affected by inorganic soil N in the absence of N fertilizer. Thus an estimate of the amoutns of N fixed could be obtained from simple estimates of legume biomass provided tissue levels of other nutrients such as phosphorus and potassium are adequate. Key factors for the achievement of sustainable grass/legume pastures include the selection of appropriate germplasm adapted to the particular environment and the judicious use of fertilizers such as phosphorus, potassium, calcium, magnesium and sulphur on acid infertile soils typical of the sub-humid and humid tropics. The main constraints to the widespread adoption of forage legumes include a lack of legume persistence, the presence of anti-quality factors such as tannins, variable Bradyrhizobium requirements and lack of acceptability by farmers. Strategies for the alleviation of these constrainst are discussed. Forage legumes can be used to recuperate degraded soils via their ability to improve the physical, chemical and biological properties of soils and these benefits could be of particular use for small-scale resource-poor farmers. The incorporation of forage legumes into agropastoral systems is discussed as an environmentally and economically attractive means to encourage the widespread adoption of legumes in the humid tropics.     

The use of carbon isotope ratios to evaluate legume contribution to soil enhancement in tropical pastures

Soil carbon distribution with depth, stable carbon isotope ratios in soil organic matter and their changes as a consequence of the presence of legume were studied in three 12-year-old tropical pastures (grass alone —Brachiaria decumbens (C₄), legume alone —Pueraria phaseoloides (C₃) and grass + legume) on an Oxisol in Colombia. The objective of this study was to determine the changes that occurred in the¹³C isotope composition of soil from a grass + legume pasture that was established by cultivation of a native savanna dominated by C₄ vegetation. The¹³C natural abundance technique was used to estimate the amount of soil organic carbon originating from the legume. Up to 29% of the organic carbon in soil of the grass + legume pasture was estimated to be derived from legume residues in the top 0–2-cm soil depth, which decreased to 7% at 8–10 cm depth. Improvements in soil fertility resulting from the soil organic carbon originated from legume residues were measured as increased potential rates of nitrogen mineralization and increased yields of rice in a subsequent crop after the grass + legume pasture compared with the grass-only pasture. We conclude that the¹³C natural abundance technique may help to predict the improvements in soil quality in terms of fertility resulting from the presence of a forage legume (C₃) in a predominantly C₄ grass pasture.     

The effect of the presence of a forage legume on nitrogen and carbon levels in soils under Brachiaria pastures in the Atlantic forest region of the South of Bahia,Brazil

The impact of forest clearance, and its replacement by Brachiaria pastures, on soil carbon reserves has been studied at many sites in the Brazilian Amazonia, but to date there appear to be no reports of similar studies undertaken in the Atlantic forest region of Brazil. In this study performed in the extreme south of Bahia, the changes in C and N content of the soil were evaluated from the time of establishment of grass-only B. humidicola and mixed B. humidicola/Desmodium ovalifolium pastures through 9 years of grazing in comparison with the C and N contents of the adjacent secondary forest. The decline in the content of soil C derived from the forest (C3) vegetation and the accumulation of that derived from the Brachiaria (C4) were followed by determining the ¹³C natural abundance of the soil organic matter (SOM). The pastures were established in 1987, 10 years after deforestation, and it was estimated that until 1994 there was a loss in forest-derived C in the top 30 cm of soil of approximately 20% (9.1 Mg C ha^–1). After the establishment of the pastures, C derived from Brachiaria accumulated steadily such that at the final sampling (1997) it was estimated 13.9 Mg ha^–1 was derived from this source under the grass-only pasture (0–30 cm). Samples taken from all pastures and the forest in 1997 to a depth of 100 cm showed that below 40 cm depth there was no significant contribution of the Brachiaria-derived C and that total C reserves under the grass/legume and the grass-only pastures were slightly higher than under the forest (not significant at P=0.05). The more detailed sampling under the pastures showed that to a depth of 30 cm there was significantly (P<0.05) more C under the mixed pasture than the grass-only pasture. It was estimated that from the time of establishment the apparent rate of C accumulation (0–100 cm depth) under the grass/legume pastures (1.17 Mg ha^–1 yr^–1) was almost double that under the grass-only pastures (0.66 Mg ha^–1 yr^–1). The data indicated that newly incorporated SOM derived from the Brachiaria had a considerably higher C:N ratio than that present under the forest.     

Nitrogen fixation and nitrogen balance studies in Rhizobium-VA mycorrhiza inoculated legume-grass association by15N isotope dilution technique under a field condition

A mixed pasture comprising of buffel grass and a legume siratro was studied under field condition for a two-year period to know the fodder yield increase, nitrogen fixation and nitrogen balance with and without the inoculation of VA mycorrhiza to grass and Rhizobium to legume component.¹⁵N dilution technique was followed using labelled ammonium sulphate. The data showed that during the first year of the above study combined inoculation of VA mycorrhiza and Rhizobium to grass and legume respectively significantly increased the total dry matter (DM) (23,900 kg ha^–1 yr^–1) and total N content (308 kg ha^–1 yr^–1) of the mixed pasture over the uninoculated mixture. However, the above increase due to combined inoculation was maximum during second year with respect to DM yield (28,200 kg ha^–1 yr^–1), but the total N harvested through grass-legume mixture was comparatively lower than the first year (297 kg ha^–1 yr^–1). The amount of biologically fixed N was highest in the first year (79 kg ha^–1 yr^–1) and showed a very drastic reduction at the end of second year (39 kg ha^–1 yr^–1). A positive nitrogen balance was observed in the grass-legume mixture irrespective of inoculation of VA mycorrhiza and/or Rhizobium.     

Long- and short-term effects of crop residues on aluminum toxicity,phosphorus availability and growth of pearl millet in an acid sandy soil

Pasture swards containing perennial ryegrass (Lolium perenne L.) alone or with one of five different white clover (Trifolium repens L.) cultivars were examined for production and transfer of fixed nitrogen (N) to grass under dairy cow grazing. Grass-only swards produced 21% less than mixed clover-grass swards during the second year after sowing. Production from grass-only plots under a mowing and clipping removal regime was 44% less than from grass-only plots under grazing. Much of this difference could be attributed to N transfer. In swards without clover, the ryegrass component also decreased in favour of other grasses.The average amount of fixed N in herbage from all clover cultivars was 269 kg N ha^–1 yr^–1. Above-ground transfer of fixed N to grasses (via cow excreta) was estimated at 60 kg N ha^–1 yr^–1. Below-ground transfer of fixed N to grasses was estimated at 70 kg N ha^–1 yr^–1 by ¹⁵N dilution and was similar for all clover cultivars. Thus, about 50% of grass N was met by transfer of fixed N from white clover during the measurement year. Short-term measurements using a ¹⁵N foliar-labelling method indicated that below-ground N transfer was largest during dry summer conditions.     

Nitrogen fixation by white clover in pastures grazed by dairy cows: Temporal variation and effects of nitrogen fertilization

Effects of rate of nitrogen (N) fertilizer and stocking rate on production and N₂ fixation by white clover (Trifolium repens L.) grown with perennial ryegrass (Lolium perenne L.) were determined over 5 years in farmlets near Hamilton, New Zealand. Three farmlets carried 3.3 dairy cows ha^–1 and received urea at 0, 200 or 400 kg N ha^–1 yr^–1 in 8–10 split applications. A fourth farmlet received 400 kg N ha^–1 yr^–1 and had 4.4 cows ha^–1.There was large variation in annual clover production and total N₂ fixation, which in the 0 N treatment ranged from 9 to 20% clover content in pasture and from 79 to 212 kg N fixed ha^–1 yr^–1. Despite this variation, total pasture production in the 0 N treatment remained at 75–85% of that in the 400 N treatments in all years, due in part to the moderating effect of carry-over of fixed N between years.Fertilizer N application decreased the average proportion of clover N derived from N₂ fixation (P_N; estimated by ¹⁵N dilution) from 77% in the 0 N treatment to 43–48% in the 400 N treatments. The corresponding average total N₂ fixation decreased from 154 kg N ha^–1 yr^–1 to 39–53 kg N ha^–1 yr^–1. This includes N₂ fixation in clover tissue below grazing height estimated at 70% of N₂ fixation in above grazing height tissue, based on associated measurements, and confirmed by field N balance calculations. Effects of N fertilizer on clover growth and N₂ fixation were greatest in spring and summer. In autumn, the 200 N treatment grew more clover than the 0 N treatment and N₂ fixation was the same. This was attributed to more severe grazing during summer in the 0 N treatment, resulting in higher surface soil temperatures and a deleterious effect on clover stolons.In the 400 N treatments, a 33% increase in cow stocking rate tended to decrease P_N from 48 to 43% due to more N cycling in excreta, but resulted in up to 2-fold more clover dry matter and N₂ fixation because lower pasture mass reduced grass competition, particularly during spring.     

施硫肥对内蒙古典型草原放牧生态系统硫循环的影响及硫肥需要量的研究

应用总体平衡 (mass_balance)法研究了施硫肥 (0 ,30及 6 0kgS/hm2 )对内蒙古典型草原放牧生态系统硫循环的影响及在硫肥需要量上的应用。结果表明 ,施硫肥使牧草硫的吸收量提高了 5 0 % ,并使放牧系统硫的生物循环速率提高了 15 %以上。 1995和 1996年两年内两种硫肥处理 30和 6 0kgS/hm2 的硫的利用效率分别为 74.0 %和37.6 %。当其他硫的来源较低时 ,土壤中有机硫的矿化是草原有效硫的主要来源 ,约占整个有效硫输入量的 70 %。放牧家畜在物质循环中具有重要的生态功能 ,其硫采食量的 90 %左右以排泄物的形式返回到土壤 ,经过排泄物而释放的有效硫量约占硫的生物再循环量的 30 %。土壤中硫的淋溶损失是放牧系统中硫的主要输出形式 ;同时 ,家畜尿和粪中硫的损失 (包括转移到非生产区和淋溶损失 )也影响着放牧系统硫的平衡状况。因此 ,应该深入研究粪尿硫的再循环速率及其影响因素。基于总体平衡原则 ,该地区放牧系统中至少每年应施入 10kgS/hm2 才能保持有效硫的平衡状态     

Nitrogen cycling in grazed pastures at elevated CO2: N returns by ruminants

In pastures grazed by large herbivores, nutrients cycle both through litter and animal excreta. We compared nitrogen (N) returns from sheep grazing a temperate pasture exposed to ambient or elevated CO₂ (475 μmol mol^?1) in a FACE (Free Air CO₂ Enrichment) experiment established in the spring of 1997. In the spring of 2000 and 2001, we measured the chemical composition of the diet, sheep faeces and of individual plant species before grazing to characterize feed intake and to compare the intake of N to the N produced in faeces. In both years under elevated CO₂, leaves of the individual species exhibited lower N concentrations and higher water‐soluble carbohydrate (WSC) concentrations. There was a significantly greater proportion of legume in the diet at elevated CO₂ but, together with the changes in chemical composition of individual species, this resulted in diets that had similar N but higher WSC and digestibility for both ambient and elevated CO₂. We found that a greater proportion of dietary N was partitioned to urine at elevated CO₂, probably because of the higher proportion of legume N in the diet, with possible differences in protein quality. A potentially significant consequence of this change in partitioning is greater N loss through volatilization at higher CO₂ levels.     

Small-scale species richness and its spatial variation in an alpine meadow on the Qinghai-Tibet Plateau

We investigated how the high small-scale species richness of an alpine meadow on the Qinghai-Tibet Plateau, China, is maintained. This area is characterized by strong wind and severe cold during long winters. In winter, most livestock is grazed on dead leaves in small pastures near farmers’ residences, whereas in the short summer, livestock is grazed in mountainous areas far from farmers’ residences. The number of plant species and the aboveground biomass were surveyed for three adjacent pastures differing in grazing management: a late-winter grazing pasture grazed moderately from 1 February to 30 April, an early-winter grazing pasture grazed lightly from 20 September to late October, and a whole-year grazing pasture grazed intensively throughout the entire year. In each pasture, we harvested the aboveground biomass from 80 or 100 quadrats of 0.01 m² along a transect and classified the contents by species. We observed 15.5–19.7 species per 0.01 m², which is high richness per 0.01 m² on a worldwide scale. The species richness in the two winter grazing pastures was higher than that in the whole-year grazing pasture. The spatial variation in species richness and species composition in the two winter grazing pastures in which species richness was high was greater than that in the whole-year grazing pasture in which species richness was lower. Most of the leaves that are preserved on the winter grazing pastures during summer are blown away by strong winds during winter, and the remaining leaves are completely exhausted in winter by livestock grazing. A pasture with a high richess is accompanied by a high spatial variation in species richness and species composition. There is a high possibility that the characteristic of spatial variation is also caused by traditional grazing practices in this area.     

Nitrogen relationships in intensively managed temperate grasslands

Summary Most studies of N relationships in grassland have used cut swards. These have shown that for annual inputs of 200 to 400 kg N/ha from fertilizer or fixation, 55 to 80% of the N is recovered in harvested herbage. Generally, no more than 5 to 15% is lost through leaching and denitrification with most of the remaining N incorporated into soil organic matter. The relatively high efficiency of N use by cut swards reflects rapid uptake of N and the removal of a large part of the input in herbage. Inclusion of the grazing ruminant alters the efficiency of N use; only 5–20% of the input is recovered in meat or milk, and 75 to 90% of the N ingested is excreted, mainly as urea in urine. Application of N in urine ranges from 30–100 g/m². Too much N is voided for effective recovery by the sward whilst soils usually contain insufficient C to allow appreciable immobilization. The surfeit is lost. Hydrolysis of urea is usually complete within 24 h of urine deposition. For urine-treated pasture in New Zealand (NZ) losses by NH₃ volatilization of up to 66% of applied N are found during warm dry weather, with an average of 28% for a range of seasonal conditions. In the UK, the average rate of NH₃ loss from an intensively grazed ryegrass sward was 0.75 kg N/ha/day during a 6-month season. NH ₄ ⁺ remaining in the soil may be nitrified, nitrification being complete within 3 to 6 weeks. Although some NO ₃ ^– is recovered by plants, a substantial portion is leached and/or denitrified. On average such losses were 42%, with only 30% of the added N recovered by plants in urine-treated pasture in NZ. In the UK annual leaching of 150 to 190 kg N/ha has been observed for grazed swards receiving 420 kg N/ha/yr. Low retention of N by grazing ruminants results in a breakdown of N relationships in intensively managed grasslands. The substantial losses through NH₃ volatilization, leaching and denitrification have serious agronomic, economic and environmental implications.     

Rio Grande Wild Turkey Habitat Selection in the Southern Great Plains

ABSTRACT We recorded telemetry locations from 1,129 radiotagged turkeys (Meleagris gallopavo intermedia) on 4 study areas in the Texas Panhandle and southwestern Kansas, USA, from 2000 to 2004. Analyses of telemetry locations indicated both sexes selected riparian vegetative zones. Females did not select grazed or nongrazed pastures for daily movements. However, females did select nongrazed pastures for nest sites on 2 study areas and males selected for grazed pastures at one study area during the breeding season. We compared nest sites (n = 351) to random sites using logistic regression, which indicated height of visual obstruction, percent canopy cover, and percent bare ground provided the highest predictive power (P ≤ 0.003) for characteristics describing nest-site selection. Nest-site vegetative characteristics between vegetative zones differed primarily in composition: upland zone nest sites had more (P ≤ 0.001) shrubs and riparian zone nest sites had more (P ≤ 0.001) grass. There were no differences in measured nest site vegetative characteristics between pasture types, but there were differences between available nesting cover in grazed and nongrazed pastures. Random plots in grazed pastures had less grass cover (P ≤ 0.001) and more bare ground (P = 0.002). Because of cattle impacts on average grass height and availability, grazing would likely have the highest impact on nesting in riparian zones due to turkey use of grass as nesting cover. An appropriate grazing plan to promote Rio Grande turkey nesting habitat would include grazing upland zones in the spring, when it likely has little impact on nesting-site selection, and grazing riparian zones following breeding season completion. Grazing at light to moderate intensities with periods of rest did not affect male turkey pasture use and may have continued to maintain open areas used by male turkeys for displaying purposes.     

Nitrogen release from eucalypt leaves and legume residues as influenced by their biochemical quality and degree of contact with soil

Large areas of short-rotation eucalypt plantations are being established in south-western Australia on land previously used for agriculture. Options for maintaining soil N supply include retention of harvest residues and legume inter-cropping. We evaluated the effects of adding the residues of five legume species and Eucalyptus globulusleaves on inorganic N dynamics in two soils (a Rhodic Ferralsol or red earth and a Haplic Podzol or grey sand) using two modes of residue application in a laboratory incubation experiment (519 days). The time course of net N immobilisation and mineralisation in both soils was strongly influenced by the type and mode of application. Eucalypt leaves caused strong N immobilisation (–7 mg N g^–1 residue-C) over the entire 519-day incubation, whereas for the legume species, N that was eventually immobilised at the start of the incubation, remineralised later to different degrees. Amongst the legumes, largest amounts of N were released from lupin residues (18 mg N g^–1 residue-C) and lowest amounts from field pea (2 mg N g^–1 residue-C). However, initial residue quality parameters were not significantly (P > 0.05) correlated with N release from the residues. Grinding and incorporating of the residues caused a much greater immobilisation of N than when residues were cut and surface applied. When ground residues were incorporated, immobilisation of N was more severe and endured for longer in the finer textured red earth than in the coarse textured grey sand. Where residues were surface applied, N dynamics were similar for both soil types. The results of this study suggest that legumes used as a mulch in eucalypt plantations are a readily available source of N for trees, and that the benefits from retention of harvest residues are more likely in maintaining soil N fertility on the long-term.     

Nitrogen inputs and losses from New Zealand dairy farmlets, as affected by nitrogen fertilizer application: year one

Inputs and losses of nitrogen (N) were determined in dairy cow farmlets receiving 0, 225 or 360 kg N ha^-1 (in split applications as urea) in the first year of a large grazing experiment near Hamilton, New Zealand. Cows grazed perennial ryegrass/white clover pastures all year round on a free-draining soil. N₂ fixation was estimated (using ¹⁵N dilution) to be 212, 165 and 74 kg N ha^-1 yr^-1 in the 0, 225 and 360 N treatments, respectively. The intermediate N rate had little effect on clover growth during spring but favoured more total pasture cover in summer and autumn, thereby reducing overgrazing and resulting in 140% more clover growth during the latter period.Removal of N in milk was 76,89 and 92 kg N ha^-1 in the 0, 225 and 360 N treatments, respectively. Denitrification losses were low (7–14 kg N ha^-1 yr^-1), increased with N application, and occurred predominantly during winter. Ammonia volatilization was estimated by micrometeorological mass balance at 15, 45 and 63 kg N ha^-1 yr^-1 in the 0, 225 and 360 N treatments, respectively. Most of the increase in ammonia loss was attributed to direct loss after application of the urea fertilizer.Leaching of nitrate was estimated (using ceramic cup samplers at 1 m soil depth, in conjunction with lysimeters) to be 13, 18 and 31 kg N ha^-1 yr^-1 in a year of relatively low rainfall (990 mm yr^-1) and drainage (170–210 mm yr^-1). Drainage was lower in the N fertilized treatments and this was attributed to enhanced evapotranspiration associated with increased grass growth.Nitrate-N concentrations in leachates increased gradually over time to 30 mg L^-1 in the 360 N treatment whereas there was little temporal variation evident in the 0 (mean 6.4 mg L^-1) and 225 (mean 10.1 mg L^-1) N treatments. Thus, the 360 N treatment had a major effect by greatly reducing N₂ fixation and increasing N losses, whereas the 225 N treatment had little effect on N₂ fixation or on nitrate leaching. However, these results refer to the first year of the experiment and further measurements over time will determine the longer-term effects of these treatments on N inputs, transformations and losses.     

Estimates of seasonal nitrogen fixation of annual subterranean clover-based pastures using the15N natural abundance technique

Annual pasture legumes play a key role in ley farming systems of southern Australia, providing biologically fixed nitrogen (N) to drive the production of the pastures as well as subsequent crops grown in rotation. Seasonal inputs of biologically fixed N in shoot biomass of the subterranean clover (Trifolium subterraneum) component of grazed annual pastures were assessed using the¹⁵N natural abundance technique and appropriately timed sampling of herbage dry matter (DM) for N accumulation. At three study sites spanning a gradient across the Western Australian wheatbelt from 300 to 600 mm annual rainfall the performance of the clover and non-legume herbs and grasses was examined as paired comparisons involving two management treatments expected to give contrasting effects on pasture productivity, botanical composition and N₂ fixation. The proportion of clover N derived from atmospheric N₂ fixation (%Ndfa) ranged from 65 to 95% across sites, treatments and sampling times. Amounts of fixed N accumulated in clover shoot biomass ranged from 50 to 125 kg ha⁻¹, and paralleled trends in clover production. Substantial increases in pasture production in high yielding treatments generally occurred without decrease in %Ndfa, suggesting that N₂ fixation was essentially non-limiting to performance of the clover component. Seasonal profiles for accumulation of fixed N were skewed towards the late winter and spring period, particularly in low plant density pastures following a cereal crop. There were seasonal, site and treatment-specific effects on the proportion of clover and non-legume pasture components and consequently clover yield and N₂ fixation were variably affected by competition from non-legume species.     

Influence of biochemical quality on C and N mineralisation from a broad variety of plant materials in soil

We studied C and N mineralisation patterns from a large number of plant materials (76 samples, covering 37 species and several plant parts), and quantified how these patterns related to biological origin and selected indicators of chemical composition. We determined C and N contents of whole plant material, in water soluble material and in fractions (neutral detergent soluble material, cellulose, hemicellulose and lignin) obtained by stepwise chemical digestion (modified van Soest method). Plant materials were incubated in a sandy soil under standardised conditions (15 °C, optimal water content, no N limitation) for 217days, and CO₂ evolution and soil mineral N contents were monitored regularly. The chemical composition of the plant materials was very diverse, as indicated by total N ranging from 2 to 59 mg N g^–1, (i.e. C/N-ratios between 7 and 227). Few materials were lignified (median lignin=4% of total C). A large proportion of plant N was found in the neutral detergent soluble (NDS) fraction (average 84%) but less of the plant C (average 46%). Over the entire incubation period, holocellulose C content was the single factor that best explained the variability of C mineralisation (r=–0.73 to –0.82). Overall, lignin C explained only a small proportion of the variability in C mineralisation (r=–0.44 to –0.51), but the higher the lignin content, the narrower the range of cumulative C mineralisation. Initial net N mineralisation rate was most closely correlated (r=0.76) to water soluble N content of the plant materials, but from Day 22, net N mineralisation was most closely correlated to total plant N and NDS-N contents (r varied between 0.90 and 0.94). The NDS-N content could thus be used to roughly categorise the net N mineralisation patterns into (i) sustained net N immobilisation for several months; (ii) initial net N immobilisation, followed by some re-mineralisation; and (iii) initially rapid and substantial net N mineralisation. Contrary to other studies, we did not find plant residue C/N or lignin/N-ratio to be closely correlated to decomposition and N mineralisation.     

Biological nitrogen fixation in grass–clover affected by animal excreta

Aiming at estimating the average N2-fixation in a pasture, ap preciating the great variability due to patchy urine and dung deposition, the in fluence of dairy cow excreta on biological N2-fixation in a perennial ryegrass–white clover mixture was studied using natural urine and dung. Application of urine as well as dung affected the N2-fixation by promoting the growth of grass and thereby the proportion of clover was significantly reduced. Also the proportion of clover-N derived from the atmosphere (pNdfa) was significantly reduced. In control plots clover dry matter constituted between 40 and 50% of the total dry matter production and the pNdfa ranged between 0.8 and 0.9. Addition of urine caused a significant increase in the grass growth rates, which was the primary reason for a decrease in proportion of clover. At the same time pNdfa decreased to 0.2–0.4 followed by an increase resulting in a total reduction of 45% in the N2-fixation in urine affected areas over a period of four months. The dung only affected the N2-fixation for a distance of up to 10 cm from the edge of the dung pats. In this border area the pNdfa decreased from 0.85 to 0.75 during one month after application followed by an increase, so that after three months there was no difference between pNdfa at 0–10 and 10–20 cm distance from the dung hill. The proportion of clover was lower in the 0–10 cm than in the 10–20 cm distance, which totally resulted in a total reduction of 20% in the N2-fixation over a period of four months in the 0–10 cm area around the dung pats. Considering the proportion of a pasture which may by affected by excreta at a stocking density of 4–6 cows ha-1, the length of the grazing period, the frequency of excretion and the area covered by individual patches, it was estimated that the N2-fixation in a grass-clover pasture would be reduced by 10–15% compared to the N2-fixation in a grass-clover sward not exposed to animal excreta.     

Effects of restoration with cattle grazing on plant species composition and richness of semi-natural grasslands

Restoration of semi-natural grasslands by cattle grazing is among the most practical options for reversing the decline of northern European floristic diversity, but no studies on this subject are available. In this work the success of restoration of abandoned, privately owned mesic semi-natural grasslands by farmers receiving support from the EU agri-environmental support scheme was studied in southwestern Finland. Three kinds of grasslands were compared: old (continuously cattle grazed), new (cattle grazing restarted 3–8 years ago) and abandoned pastures (grazing terminated >10 years ago). Plant species composition of the three pasture types was floristically different in multivariate analyses (non-metric multidimensional scaling). Total species richness, richness of grassland plants, indicator plants and rare plants were highest in old and lowest in abandoned pastures in all studied spatial scales (0.25–0.8 ha, 1 and 0.01 m²). The results were congruent with different scales and species list definitions, suggesting that species density scale (1 m²) can be used as a partial surrogate for large scale species richness. Species richness of new pastures was 20% higher on 0.25–0.8 ha, 40–50% higher on 1 m² and 30% higher on the 0.01 m² scale compared to abandoned grasslands. Rare species showed insignificant response to resumed grazing. Despite problems in management quality, this study showed promising results of restoration of abandoned grasslands by cattle grazing on private farms. However, populations of several rare grassland plants may not recover with present cattle grazing regimes. Management regulations in the agri-environmental support scheme need to be defined more precisely for successful restoration.