Field evaluation of lentil cultivars inoculated with Rhizobium leguminosarum bv. viciae strains for nitrogen fixation using nitrogen-15 isotope dilution

 A ¹⁵N isotope dilution technique was applied to quantify the extent of N₂ fixation in lentil (Lens culinaris Medik.) cultivars as influenced by Rhizobium leguminosarum bv. viciae strains in a field experiment in Pakistan. The experiment was conducted on a soil with a very small indigenous rhizobial population and where N was a limiting factor for crop production. Significant variations in number of nodules, dry weight of nodules, biomass yield, grain yield, total N yield, proportion of plant N derived from N₂ fixation (P_fix) and amount of N derived from the atmosphere (N_dfa) were observed among combined treatments of four rhizobial strains and six lentil varieties. In a field previously labelled with ¹⁵N, to which a basal dose of 75 kg P₂O₅ ha^–1 was applied as single super phosphate, N_dfa ranged from 15 to 24 kg N ha^–1 when calculated according to rhizobial strain and from 4 to 38 kg N ha^–1 when calculated according to lentil variety. Lc 26 was the most effective strain and fixed 243% more N than the indigenous population in the uninoculated control. In treatments with the lentil variety PL-406, N_dfa was 38 kg N ha^–1, which was 850% higher than with the lentil variety Precoz/F6-20-1×M-85. Generally, the varieties with greater P_fix produced a higher dry matter yield. Received: 26 May 1999     

Effect of climate, soil type and earthworm activity on nitrogen transfer from a nitrogen-15-labelled decomposing material under field conditions

 N transfer from ¹⁵N-labelled decomposing material into the microbial biomass and inorganic N forms was studied for more than 2 years at three experimental sites differing in climatic conditions and earthworm abundance. The ¹⁵N-labelled decomposing material was mixed with low-elevation soil (LES), mid-elevation soil (MES) and high-elevation soil (HES). The amended soils were put into two kinds of plastic cylinders closed on both sides with nets preventing (0.1 cm mesh) and allowing (0.5 cm mesh) access by earthworms, and were buried in soil (20 cm depth) to monitor the transfer of N from the ¹⁵N-labelled decomposing material. Climate and soil type play an important role in the release of N from decomposing material. LES transplanted to more humid sites (mid- and high-elevation sites) showed an increase in most of its biological activities (N atom % excess, and microbial biomass C and N). Furthermore, LES was a sandy soil in which the ¹⁵N-labelled decomposing material was less bound than in MES and HES, which contained more silt and clay. This resulted in faster organic matter turnover when climatic conditions were favourable. The presence of earthworms greatly increased the quantity of inorganic N (mainly NH₄ ⁺) in the soils and enhanced the release of N from the ¹⁵N-labelled decomposing material and the native organic matter, compared to soil without earthworms. Received: 21 January 1999     

The use of nitrogen-15 natural abundance in white clover (Trifolium repens L.) to determine nitrogen fixation under different management practices

 The ¹⁵N natural abundance (δ¹⁵N) of white clover (Trifolium repens L.) grown in pasture under different management practices was determined. Plants were split into leaflets, petioles and stolons and the ¹⁵N signature of each tissue was measured. The δ¹⁵N of leaflet tissue from plants of two non-N₂-fixing species (Lolium perenne L. and Ranunculus repens L.), growing in close proximity to the sampled T. repens, was also measured. By using T. repens plants grown in the absence of mineral N to provide reference material, the proportion of N derived from N₂ fixation (%Ndfa) in pasture plants was calculated. Within a plot, variation was present in the δ¹⁵N between the tissues of T. repens. Variation was also present between the same tissues under different management practices. The %Ndfa in the leaf material of T. repens varied from 34% to 100% between the plots. The use of different reference species did not affect the estimate of %Ndfa. Received: 14 December 1998     

采用15N同位素稀释法研究不同层次土壤氮素总转化速率

采用15N同位素稀释方法,开展短期(7天)室内培养实验,估算了一水稻土0~20、20~60和60~90 cm土层土壤主要N素转化过程的总转化速率,结果表明,标记N溶液加入后2 h内各土层土壤的总矿化、硝化、固定速率显著高于其他时间段(p<0.01)。2 h后,矿化速率在小范围内起伏。0~20 cm土层土壤N素的硝化速率随培养时间延长而降低,另外两层土壤则基本保持稳定,硝化速率的变化与硝化作用底物NH4+-N浓度的变化呈显著正相关。值得注意的是,外源无机N溶液加入后2 h内,大量NH4+-N和NO3--N被固定,并认为N素的非生物固定起主导作用。2 h后,出现了N素在固定与再矿化间反复转换的现象。实验结果表明,与净转化速率相比总转化速率能更好地描述单个N素转化过程,但由于外源N加入对N素转化的影响、再矿化作用以及忽略了N素转化过程中的气体损失、DNRA(硝态氮异化还原为铵)过程等,本研究结果与真实值间存在一定差异。     

The use of 15N labelling to study the turnover and utilization of ruminant manure N

 An improved understanding of the cycling of animal manure N is a prerequisite for making better use of this N source. A sheep was fed ¹⁵N-labelled grass in order to study the fate of ¹⁵N-labelled ruminant manure N in the plant-soil system. The uniformity of labelling was found to be satisfactory when an appropriate feeding strategy was used. The mineralization of labelled faecal N was compared to the mineralization of labelled feed N and indigestible feed N by measuring residual labelled organic N in unplanted topsoil in the field. After 18 months, 61% of both faecal N and feed N was recovered in organic form in the topsoil, while 94% of the indigestible feed N was still present in the soil. The influence of slurry distribution in soil on the crop uptake of labelled faecal N in slurry was studied in a sandy and a sandy loam soil. The crop uptake of labelled faecal N was compared with the uptake of ¹⁵N-labelled mineral fertilizer in a reference treatment. The uptake was 28–32% of that of the reference treatment with simulated slurry injection, 13–25% with incorporated slurry and 18–19% with slurry on the soil surface. The mineralization of faecal N in the autumn after application in spring was low irrespective of the slurry distribution in soil. The results demonstrate that the contact between animal manure and the soil matrix significantly influences the short-term turnover and availability of faecal and ammonium N in slurry, especially in fine-textured soils. Received: 31 October 1997     

Genetic variability in dinitrogen fixation between cowpea [Vigna unguiculata (L.) Walp] cultivars determined using the nitrogen-15 isotope dilution technique

 N fixed in 16 cultivars of cowpea [Vigna unguiculata (L.) Walp] inoculated with effective Bradyrhizobium strains collected from the West African MIRCEN culture collection was measured by ¹⁵N isotope dilution technique. In all plant parts, significant differences in the percentage of N derived from the atmosphere (%Ndfa) and the amount of Ndfa occurred between the cultivars. Ndoute variety exhibited the highest %Ndfa (74.33% in shoots; 60.90% in roots) and accumulated more fixed N (960 mg N plant^–1 and 38 mg N plant^–1 in shoots and roots, respectively). Therefore this cultivar should be selected as the highest N-fixing cowpea cultivar. It also should be used in a breeding programme to contribute to the development of cultivars that could stimulate an intensive use of cowpea in many different cropping systems in Africa with a view to maintaining soil fertility. Received: 14 June 1999     

Estimation of simultaneous nitrification and denitrification in grassland soil associated with urea-N using 15N and nitrification inhibitor

 A low efficiency of use of N fertilisers has been observed in mid-Wales on permanent pasture grazed intensively by cattle. Earlier laboratories studies have suggested that heterogeneity in redox conditions at shallow soil depths may allow nitrification and denitrification to occur concurrently resulting in gaseous losses of N from both NH₄ ⁺ and NO₃ ^–. The objective of the investigation was to test the hypothesis that both nitrification and denitrification can occur simultaneously under simulated field capacity conditions (∼5 kPa matric potential). Intact soil cores were taken from grassland subjected to both grazing and amenity use. The fate of applied NH₄ ⁺ was examined during incubation. ¹⁵N was used as a tracer. Nitrapyrin was used as a nitrification inhibitor and acetylene was used to block N₂O reductase. More than 50% of N applied as NH₄ ⁺ disappeared over a period of 42 days from the soil mineral-N pool. Some of this N was evolved as N₂O. Accumulation of NO₃ ^––N in the surface 0–2.5 cm indicated active nitrification. Addition of nitrapyrin increased N recovery by 26% and inhibited both the accumulation of NO₃–N and emission of N₂O. When intact field cores were incubated after addition of ¹⁵N-urea, all of the N₂O evolved was derived from added urea-N. It was concluded that nitrification and denitrification do occur simultaneously in the top 7.5 cm or so, of the silty clay loam grassland topsoils of mid-Wales at moisture contents typical of field capacity. The quantitative importance of these concurrent processes to N loss from grassland systems has not yet been assessed. Received: 15 December 1998     

Uptake of 15N by wetland rice in response to application of 15N-labelled Sesbania rostrata and urea

 Pot experiments were carried out to evaluate the response of rice to Sesbania rostrata green manure N as compared to urea fertilizer N under flooded conditions. After growing S. rostrata for 21 days with a ¹⁵N-labelled N source, the labelled Sesbania was applied to wetland rice as a green manure and the uptake of ¹⁵N from this substrate was compared to that from labelled urea. Rice was cultivated twice in the same pots. The rice was grown for a period of 49 days in each case, separated by a period of 21 days when the soil was allowed to dry. The ¹⁵N content of the soil and shoots and roots of rice was determined and ¹⁵N balances established. The total N content of the shoots and roots of rice was determined by a non-tracer method. The percentage recovery of ¹⁵N from shoot material which was derived from urea N was more than twice that from S. rostrata. The recovery of ¹⁵N from the pots receiving both green manure and urea was low, and not significantly different from that recovered from the green manure treatment. As much as 64.5–73.5% and 40.1–41% of the ¹⁵N remained in the soil which had received green manure or urea, respectively. The overall recoveries of ¹⁵N varied between 86.5% and 94.4%. At the second harvest, the oven-dry weight of shoots was significantly (P<0.05) higher in green-manure treated pots, but the total N content did not differ significantly. Labelled N remaining in the soil after amendment with the green manure was much more available to the rice crop than that remaining after the addition of urea-N. The total recovery of labelled N (shoots plus roots) amounted to 65.5% and 74%, respectively of the residual labelled N in the two S. rostrata treatments (i.e. 19.55 mg ¹⁵N pot^–1 and 39.10 mg ¹⁵N pot^–1) and 23.2% and 23.2% of the residual labelled N in the two urea treatments (i.e. 19.55 mg ¹⁵N pot and 39.10 mg ¹⁵N pot^–1), respectively. Received: 8 December 1997     

Transformations of nitrogen-15-labelled urea in a flooded soil as affected by floodwater algae and green manure in a growth chamber

 The effects of floodwater algae and green manure on transformations of ¹⁵N-urea were studied in columns of a sandy loam soil in a growth chamber. The columns were flooded and either kept in the light, to allow algal growth, or in the dark (control) for 17 days before adding the labelled urea. Changes in urea-, NO₃ ^–- and NH₄ ⁺-N levels and the pH of the floodwater were measured over the subsequent 41-day period, during which the control column remained in the dark and those containing algae were maintained either in the dark to cause the death of the algae or in the light. Volatilized NH₃ was monitored, and on termination of the experiment the distribution of ¹⁵N between NO₃ ^–, NH₄ ⁺ and organic forms was measured in the soil. Urea hydrolysis was most rapid in the presence of both living algae and green manure, followed by dead algae, and was slowest in the control. The concentration of NH₄ ⁺-N in the floodwater was, however, reduced in the presence of algae due to assimilation and NH₃ volatilization owing to the raised day-time pH in the floodwater. NH₃ volatilization for the first 10 days was rather high in the columns kept in the light compared to those in the dark. Total volatilization plus denitrification losses were greatest where dead algae were present, owing to the absence of live algae which assimilated more than half of the applied N. Algal growth in floodwater increased the depth of the aerobic soil layer present at the soil-water interface. Subsequently, under dark conditions, stimulated algal growth reduced the depth of the aerobic layer causing less nitrification, which resulted in lower losses of N due to denitrification, i.e. 17% of the applied urea-N as compared to 39% in the light treatments. Although the presence of green manure caused a marked increase in the rate of hydrolysis, algal assimilation prevented excessive N losses via volatilization, indicating that the retention of higher quantities of NH₄ ⁺-N may have increased fertilizer-N use efficiency. Received: 22 January 1999     

Soil organic matter transformations induced by Hieracium pilosella L. in tussock grassland of New Zealand

 To study the effect of Hieracium pilosella L. invasion on the transformations of soil organic matter of New Zealand tussock grassland soils (Ustochrepts), plant material and soils underneath Hieracium, the surrounding halo, and the adjacent herbfield (depleted tussock grassland) were examined for their chemical composition. An attempt was made to reveal possible changes in chemical composition of the soil organic matter induced by H. pilosella invasion. Small differences were detected by solid-state ¹³C nuclear magnetic resonance (NMR) spectroscopy in the composition of the plant and soil materials from these zones. Most of the differences in soil organic matter occurred due to differences in the amount and quality of plant-residue inputs. Comparable amounts of phenolic C were detected in the solid-state ¹³C NMR spectra of H. pilosella and herbfield vegetation, while alkaline CuO oxidation yielded considerable lower lignin oxidation products for H. pilosella. A slightly higher proportion of these compounds in H. pilosella soil revealed an accumulation and a low degradation rate of lignin compounds under H. pilosella. The HCl hydrolysis and solid-state ¹⁵N NMR spectroscopy showed similar chemical compositions of the N fractions of the three different soils. The absence of ¹⁵N NMR signal intensity assignable to aniline derivatives or aromatic heterocyclic N indicates that the condensation of phenolic compounds with N groups plays a minor role in N sequestration in these soils. Received: 6 September 1999     

The use of 15N natural abundance variation to examine plant and soil organic fractions in pasture under different management practises

 Pasture systems lack the complexity of natural grasslands and have undisturbed soil profiles relative to arable monocultures. With controlled nitrogen (N) fertilizer inputs and measurable outputs (e.g. grazing and leaching), they can be used to investigate ¹⁵N natural abundance variation as a tool for the study of soil-N processes. In the present study, four pastures of different sward composition and N inputs were examined. Plant shoots and a range of soil fractions, categorized by size, were sampled in May prior to any major N additions, and again in July after initial N inputs had of been made. Samples were analyzed for ¹⁵N natural abundance (δ¹⁵N) and total N (εN). In the May sample plant and soil fractions varied in both ¹⁵N and εN between treatments. The 0.5 mm and 0.2 mm soil factions were comparable within treatments, as were the silt and clay fractions. Between May and July changes were apparent in the δ¹⁵N and εN of shoots and some soil fractions within each plot these corresponded to N inputs or sward type. Changes in silt-N especially, were similar to those occurring in the shoots. No comparable changes were seen in the larger fractions. Not all measured variation was explicable in this study. The inadequacies of the approach are highlighted and suggested improvements discussed. Received: 9 February 1998     

Soil N dynamics in a natural calcareous grassland under a changing climate

 This paper reports the results from a medium-term field scale investigation into the effects of simulated climate change on soil N mineralisation in a semi-natural calcareous grassland in southern England. The experiment utilised soil warming cables, automatic rainshelters and a watering system to examine two climate change scenarios: warmer winters with summer drought and warmer winters with enhanced summer rainfall. Gross N mineralisation rates in treated plots were determined, using ¹⁵N pool dilution techniques, at 6-weekly intervals over a 3-year period. Results from control plots showed a strong seasonality of mineralisation with highest rates in autumn and winter and lowest rates in summer. They suggest that water availability is the main constraint on microbial processes and plant growth. Unexpectedly, additional summer rainfall had no direct effect on N mineralisation at the time of application (summer). The treatment did, however, significantly (<0.05%) reduce rates in subsequent autumn and winter months. In contrast, summer drought significantly increased N mineralisation rates in autumn and winter. Winter warming similarly had no direct effect on N mineralisation in winter but decreased rates in spring. We hypothesise that the observed treatment effects result from changes in organic C and N input, in plant litter, resulting from the direct impact of climatic manipulation on perennial plant growth, death and senescence. This paper compares and contrasts the response to climate manipulation in the grassland system with results from other ecosystem types such as northern forests. Received: 1 December 1997     

Comparison of different methodologies for field measurement of net nitrogen mineralization in pasture soils under different soil conditions

 Net mineralization was measured in free-draining and poorly drained pasture soils using three different field incubation methodologies. Two involved the use of enclosed incubation vessels (jar or box) containing C₂H₂ as a nitrification inhibitor. The third method confined soil cores in situ in an open tube in the ground, with an anion-exchange resin at the base to retain leached NO₃ ^– (resin-core technique, RCT). Measurements were made on three occasions on three free-draining pastures of different ages and contrasting organic matter contents. In general, rates of net mineralization increased with pasture age and organic matter content (range: 0.5–1.5 kg N ha^–1 day^–1) and similar rates were obtained between the three techniques for a particular pasture. Coefficients of variation (CVs) were generally high (range: 10.4–98.5%), but the enclosed incubation methods were rather less variable than the RCT and were considered overall to be the more reliable. The RCT did not include C₂H₂ and, therefore, newly formed NO₃ ^– may have been lost through denitrification. In a poorly drained pasture soil, there were discrepancies between the two enclosed methods, especially when the soil water content approached field capacity. The interpretation of the incubation measurements in relation to the flux of N through the soil inorganic N pool is discussed and the drawbacks of the various methodologies are evaluated. Received: 18 November 1999     

Land use effects on labile N extracted with permanganate and the nitrogen management index in the Cerrado region of Brazil

 The effect of land use on the availability of nitrogen (N) was studied by separating total N into a labile and a stable fraction by oxidation and extraction of labile N with KMnO₄. The nitrogen management index (NMI) was calculated following the method of Blair et al. (1995) for the carbon management index. In all systems, labile N released by KMnO₄ was a better indicator of N availability than total and stable N. The NMI was a good indicator of N availability but gave no information about the total amount of N. In land use system analysis, total N and labile N can be used together as a simple and rapid way to evaluate the N status of the soil. Legume-based pastures specifically increased the amount of labile N. Although soybeans had a dominant role in the continuous cropping systems studied, the total N contents of these systems were lower than those of the natural Cerrado. The availability of N under legume-based pastures and legume-based pasture-crop rotations was higher than under the natural Cerrado and the continuous cropping systems. Received: 11 June 1997