Dynamics of nodulation,nitrogen fixation,nitrogen use and biomass yield over time in pot-grown Leucaena leucocephala (Lam.) de Wit

The dynamics of nodulation, N₂-fixation and N use in Leucaena leucocephala cv. K28 over time was investigated in a screenhouse at 4, 8, 12 and 16 months after planting (MAP) using the ¹⁵N-labelling method. Leucaena had a consistently increasing pattern of nodulation, dry biomass and nitrogen yield. A sharp rise in nodulation was observed between 12 and 16 MAP, whereas for biomass, N accumulation and N₂-fixation, and N₂-fixation, an upward surge occurred between 4 and 12 months. Nodulation, N accumulation, N₂-fixation and biomass yield all peaked at 16 MAP. Along with the steady increase in N₂-fixation throughout the 16-month growth period, the % N derived from the atmosphere rose from 17.9% to 61.5%, 70.1% and 74%, equivalent to 191, 1623, 2395 and 3385 mg N₂ fixed plant^-1 at 4, 8, 12 and 16 MAP, respectively. Nitrogen assimilation from soil and fertilizer decreased inversely to the increase in symbiotic nitrogen fixation with time.     

Effects of NPK fertilizer combinations on yield and nitrogen balance in sorghum or pigeonpea on a vertisol in the semi-arid tropics

A long-term experiment was carried out on a Vertisol from 1986 to 1992 to examine the combined effects of NPK fertilizers on yield using sorghum (Sorghum bicolor L. Moench cv. CSH 5) and short-duration pigeonpea (Cajanus cajan L. Millsp. cv. ICPL 87). The fertilizer treatments were as follows: 0 (no fertilization), N (150 kg N ha^-1 ), P (65.5 kg P₂O₅ ha^-1), K (124.5 kg K₂O ha^-1), and all possible combinations (NP, NK, PK, and NPK). In this study we continued this experiment during the period 1993 to 1994 and analyzed the crop yield response to fertilizers and the N balance. The amount of N derived from the atmosphere and fertilizer was estimated by the ¹⁵N natural abundance method and ^l5N isotope dilution method, respectively. A combined application of Nand P fertilizers gave the highest grain yield for the two crops under the 8th and 9th continuous croppings, unlike the application of K fertilizer. The values of total N for the two crops were significantly higher in the NP and NPK plots. These crops took up N mainly from soil. There was a significant positive relationship between the uptake of N_dff and N_dfs by each crop. Pigeonpea or sorghum took up more N from the soil in the N fertilizer plots than in the plots without N, suggesting that soil N fertility was enhanced and the amount of N supplied from soil increased in the plots with consecutive application of N fertilizer for 7 y. Even pigeonpea, which fixes atmospheric N inherently, needed N fertilizer to achieve high grain yield, suggesting that N fixation by the nodules was not always sufficient to meet the N requirements of the crop under these conditions. Although fertilizer N exerted a beneficial effect on plant growth and yield in the two crops, the values of fertilizer N recovery (FNR) by the two crops were considerably low. Therefore, it is suggested that the development of N fertilizer management which could maximize FNR of each crop should be promoted.     

Real-time imaging of nitrogen fixation in an intact soybean plant with nodules using 13N-labeled nitrogen gas

Abstract

Real-time images of nitrogen fixation in an intact nodule of hydroponically cultured soybean (Glycine max [L] Merr.) were obtained. In the present study, we developed a rapid method to produce and purify ¹³N-labeled radioactive nitrogen gas (half life: 9.97?min). ¹³N was produced from a ¹⁶O (p, α) ¹³N nuclear reaction. The target chamber was filled with CO₂ and irradiated for 10?min with protons at an energy of 18.3?MeV and an electric current of 5?μA, which was delivered from a cyclotron. All CO₂ in the collected gas was absorbed and removed with powdered soda-lime in a syringe and replaced with helium gas. The resulting gas was injected into gas chromatography and separated and a 35?mL fraction, including the peak of [¹³N]-nitrogen gas, was collected by monitoring the chromatogram. The obtained gas was mixed with 10?mL of O₂ and 5?mL of N₂ and used in the tracer experiment. The tracer gas was fed into the underground part of intact nodulated soybean plants and serial images of the distribution of ¹³N were obtained non-invasively using a positron-emitting tracer imaging system (PETIS). The rates of nitrogen fixation of the six test plants were estimated to be 0.17?±?0.10?μmol N₂?h^?1 from the PETIS image data. The decreasing rates of assimilated nitrogen were also estimated to be 0.012?±?0.011?μmol?N₂?h^?1. In conclusion, we successfully observed nitrogen fixation in soybean plants with nodules non-invasively and quantitatively using [¹³N]N₂ and PETIS.     

Contribution of red alder to soil nitrogen input in a silvopastoral system

Studies on Alnus rubra Bong. (red alder) were carried out to assess its potential for use as a component of a silvopastoral system. Comparison was between two treatments: red alder at 400 stems ha⁻¹ (silvopasture) and red alder at 2,500 stems ha⁻¹ (forestry control). δ¹⁵N values close to zero were recorded in all red alder plant parts except for root nodules, indicating that a large proportion of N in red alder was fixed from the atmosphere. Overall, it was estimated that there was 63.45 kg N ha⁻¹ fixed N accumulated in red alder trees, and the rate of N fixation was estimated at 30.95 kg ha⁻¹ year⁻¹ in the silvopasture treatment. The total amount of N that could potentially be added to the soil in the silvopasture treatment as a result of decomposition of senescent leaves, roots, and dead nodules was estimated at 40.56 kg ha⁻¹ year⁻¹. Of the total N added to the soil, 27.1 kg ha⁻¹ year⁻¹ was due to N fixation from the atmosphere. These results show that red alder has a potential to improve and maintain soil fertility in a silvopastoral system.     

Estimating N2 fixation by field-grown lupins (Lupinus angustifolius L.) using soil and plant 15N enrichment

Summary Biological N₂ fixation was estimated in a field experiment following the addition of NH₄Cl or KNO₃ to unconfined microplots (1.5 m²) at 2.5 g N m^-2 (10 atom% ¹⁵N). A model of total N and ¹⁵N accumulation in lupins and decreasing ¹⁵N enrichment in the KCl-extractable soil-N pool (0–0.15 m depth) was used to estimate the proportion of N in lupins derived from biological N₂ fixation. Estimates of N₂ fixation derived from the model were compared with ¹⁵N isotope-dilution estimates obtained using canola, annual ryegrass, and wheat as nonfixing reference plants. Biomass, total N accumulation, or ¹⁵N enrichment in the lupin and reference crops did not differ whether NH _inf4 ^sup+ or NO _inf3 ^sup- was added as the labelled inorganic-N source. The decrease in soil ¹⁵N enrichment was described by first-order kinetics, whereas total N and ¹⁵N accumulation in the lupins were described by logistical equations. Using these equations, the uptake of soil N by lupins was estimated and was then used to calculate fixed N₂. Estimates of N₂ fixation derived from the model increased from 0 at 50 days after sowing to a maximum of 0.79 at 190 days after sowing. Those based on the ¹⁵N enrichment of the NO _inf3 ^sup- pool were 10% higher than those based on the mineral-N pool. ¹⁵N isotope-dilution estimates of N₂ fixation ranged from 0.37 to 0.55 at 68 days after sowing and from 0.71 to 0.77 at 190 days after sowing. Reference plant-derived values of N₂ fixation were all higher than modelled estimates during the early states of growth, but were similar to modelled estimates at physiological maturity. The use of the model to estimate N₂ derived from the atmosphere has the intrinsic advantage that the need for a non-fixing reference plant is avoided.     

Comparison of 15N natural abundance and difference methods for estimating N2 fixation of soybeans grown in a tropical soil

Abstract

A study was carried out to compare the difference or N-yield method with the ¹⁵N natural abundance method for the estimation of the fractional contribution of biological N₂ fixation in the different plant parts of nodulating and non-nodulating isolines of soybeans. The results indicated that the δ¹⁵N values of most plant parts of soybeans were significantly lower (p<0.05) in the nodulating than in the non-nodulating isoline. However, in the case of the root+nodule component, the δ¹⁵N value was higher in the nodulating than in the non-nodulating isoline possibly due to isotopic discrimination of ¹⁵N over ¹⁴N which may have occurred in the nodules. Inoculation of soybeans with the Bradyrhizobium japonicum strain CB 1809 increased significantly (p<0.05) the δ¹⁵N value of the root+nodule component implying that the effectiveness of the soybean-rhizobium symbiosis had increased by inoculation.

Percentage of plant N derived from atmospheric N₂ fixation (%Ndfa) estimated by the ¹⁵N natural abundance method was highly correlated (r=0.762, p<0.01) with that by the difference or N-yield method and the differences between the two methods were not statistically significant. The agreement between the two methods was closer at maturity than at the early reproductive stage.

The %Ndfa obtained by the difference method ranged from 48.4 to 92.6% whereas the %Ndfa obtained by the ¹⁵N natural abundance method ranged from 43.2 to 92.4% in the different plant parts. Based on the ¹⁵N natural abundance method, approximately 15% of the N in pod, shoot, grain, and shell was derived from the soil but in the case of stover, this fraction was about 55%.     

Nitrogen fixation in six forage legumes in Mediterranean central Chile

Abstract

The contribution of biological nitrogen fixation (BNF) to the N nutrition of six annual forage legumes, subterranean clover (Trifolium subterraneum), burr medic (Medicago polymorpha), balansa clover (Trifolium michelianum), Persian clover (Trifolium resupinatum), yellow serradela (Ornithopus compressus), and pink serradela (Ornithopus sativus) was evaluated by the ¹⁵N natural abundance technique, using four grass species (Briza máxima, Bromus mollis, Hordeum berteroanum, Avena barbata) and two composite species (Leontodon leysseri and Hedipnois cretica) as reference plants. An additional objective was to determine whether alternative legume species to those in common use (T. subterraneum and M. polymorpha) in the area, could improve BNF. The field studies were conducted in two edaphic conditions, granitic (Entisol) and clay (Vertisol) soil, located in Cauquenes, VII Region, in the sub-humid Mediterranean zone of Chile. In the granitic soil the percentages of N derived from fixation were high in all species (74 to 94%); yellow serradela cv. Tauro presented the greatest N content in dry matter and N fixation, equivalent to 91 kg N ha^?1. In contrast, pink serradela cv. Cádiz and subterranean clover cv. Gosse presented the lowest N fixation. In the clay soil, under periodically waterlogged conditions, balansa clover cv. Paradana and persian clover cv. Prolific had high percentage values of BNF (>95%) and fixed more N (100.2 and 82.5 kg N ha^?1, respectively) than burr medic and subterranean clover cv. Gosse. The present study allowed the identification of new germplasm of high capacity of N fixation which is an additional criterion for selecting species for infertile and waterlogged soil conditions in the Mediterranean area of Chile.     

Field evaluation of nitrogen fixation and use of nitrogen fertilizer by sorghum/pigeonpea intercropping on an Alfisol in the Indian semi-arid tropics

A field experiment was conducted to obtain the N balance sheet for sole crops and intercrops of sorghum [Sorghum bicolor (L.) Moench] and pigeonpeas [Cajanus cajan (L.) Millsp.]. Intercropping gave a significant advantage over sole cropping in terms of dry matter production and grain yield, as calculated on the basis of the land equivalent ratio and area-time equivalent ratio. The N fertilizer use efficiency and atmospheric N₂ fixation by pigeonpea were estimated using ¹⁵N-labeling and natural abundance methods. The N fertilizer use efficiency of sorghum was unaltered by the cropping system, while that of the pigeonpea was greatly reduced by intercropping. Although intercropping increased the fractional contribution of fixed N to the pigeonpeas, no significant difference was observed between the cropping systems in total symbiotically fixed N. There was no evidence of a significant transfer of N from the pigeonpea to the sorghum. This study showed that use of soil N and fertilizer N by pigeonpeas was almost the same as that by sorghum in sole cropping, indicating the potential competence of pigeonpeas to exploit soil N. However, when N was exhausted by a companion crop in intercropping, the pigeonpea crop increased its dependency on atmospheric N₂ fixation. We conclude that knowledge of how N from different sources is shared by companion crops is a prerequisite to establishing strategies to increase N use, and consequently land productivity, in intercropping systems.     

Quantification of the contribution of biological nitrogen fixation to Cratylia mollis using the N natural abundance technique in the semi-arid Caatinga region of Brazil

This study was performed to investigate the capacity of the woody perennial Cratylia mollis, a legume endemic to the semi-arid region of the North-East of Brazil, to nodulate, and obtain N from BNF using the ¹⁵N natural abundance technique. To estimate the ¹⁵N abundance of the N-derived from soil, the leaves of several (4-6) non-legume and non-nodulating legume species growing in close proximity to the Cratylia were analysed for δ¹⁵N. The δ¹⁵N values of these reference plants were high (from +9 to +16‰) and showed relatively small differences between species at each site/sampling time. At the irrigated site at both samplings, and at the non-irrigated site sampled in the rainy season, the ¹⁵N abundance of the Cratylia mollis leaves was far lower (+1 to +5‰) strongly suggesting that the legume obtained large proportional contributions from BNF. As was to be expected, no nodules were found on the Cratylia plants at the non-irrigated site in the dry season, and the ¹⁵N abundance of the Cratylia mollis plants were very similar to that of the reference plants, consistent and nodulation and BNF being limited by water deficit at this time.     

Soil N dynamics and N yield of barley grown on Breton loam using N from biological fixation or fertilizer

Summary Soil N dynamics and barley yields (Hordeum vulgare L.) were compared in pot experiments using surface samples from a Gray Luvisol under three cropping systems at Breton, Alberta: (1) an agroecological 8-year rotation including cereals, forage, and fababeans (Vicia faba L.) as green manure, from wich two plots were selected, one following fababeans, and the second following 3 years of forage; (2) a continuous grain system, with fertilizer N at 90 kg ha^-1 year^-1; and (3) a classical Breton 5-year rotation [following oats (Avena sativa L.)] involving forage and cereals, without returning crop residues to the land, selecting one plot with PKS treatment and a second as control. The fertilizer N equivalent for the cropping system; A_N value and A value (analogous to A_N), but in fertilizer ¹⁵N units, soil biomass, and C and N mineralization were monitored. In the first agroecological plot (after fababeans), grain and total plant biomass production were 116% greater than from the continuous grain treatment. Barley plants in the two agroecological plots derived 48.5% and 37.8%, respectively, of their N requirement from non-labelled soil N sources not present in the continuous grain plot. At crop maturity, the recovery of ¹⁵N microbial biomass was 1.5 times higher in soil from the first agroecological plot than from the continuous grain plot. The fertilizer N equivalent was 2670 mg pot^-1 (485 kg ha^-1) for the first and 1850 mg pot^-1 for the second agroecological treatment. Fertilizer N equivalent values exceed net amounts of N mineralized by a factor of 4. Recovery by the barley crop of ¹⁵N added at 55 mg pot^-1 was more efficient in the agroecological treatments (45%–51%) than in the continuous grain or classical Breton treatments (35%–37%). It was concluded (1) that past soil history may be associated more with the ability of barley plants to compete for available N, and hence the use of N, than with net soil N mineralization; and (2) an increased supply of N to crops following the incorporation of fababean residues, manure application, and the soil N-conserving effect of growing legumes were all partly responsible for the observed differences in soil fertility.     

玉米/大豆、玉米/花生间作对作物氮素吸收及结瘤固氮的影响

禾本科与豆科作物间作具有显著的增氮作用。为探明玉米/大豆、玉米/花生间作模式的氮素吸收、氮营养竞争能力及豆科结瘤特性的变化,解释玉米与豆科间作体系的增氮效应,通过田间试验,设置玉米单作（MM）、大豆单作（SS）、玉米/大豆间作（MS）、花生单作（PP）、玉米/花生间作（MP）等5种种植模式,研究不同种植模式对作物氮素积累、氮营养竞争强弱及豆科结瘤固氮特性的调控作用。结果表明,与单作相比,间作显著降低玉米和大豆的氮素积累量,对花生的氮素积累量影响不显著。5种模式系统氮素积累总量表现为MS > SS > MP,PP和MM处理最低且差异不显著,MS处理比MP处理显著高21.8%。与MM处理相比,MS和MP处理的玉米氮素积累量分别降低20.5%和11.7%,其中MP处理籽粒、叶片和茎秆氮素积累量比MS处理高8.9%、21.2%和14.3%。与SS处理相比,MS处理的大豆氮素积累量降低28.5%,其中,中行、边行分别降低10.1%、15.4%。玉米相对大豆氮营养竞争比率表现为强（CR_ms>1）,相对花生则表现为弱（CR_mp<1）。与SS处理相比,五叶期MS处理的大豆根瘤数量显著增加,根瘤鲜重无显著差异,盛花期后根瘤数量和鲜重均显著降低;MS处理的大豆根瘤固氮酶活性均降低,且中行降低幅度更大。与PP处理相比,开花期MP处理的花生根瘤数量和鲜重均显著增加,下针期后均显著降低;MP处理的花生根瘤固氮酶活性均降低,且边行降低幅度更大。各间作模式作物的氮素积累量虽然降低,但间作模式的系统氮素积累量却显著高于各单作模式,两种间作模式中MS处理的氮素积累总量最高。     

Quantitative analysis of the initial transport of fixed nitrogen in nodulated soybean plants using 15N as a tracer

The quantitative analysis of the initial transport of fixed isotope 15-nitrogen (¹⁵N) in intact nodulated soybean plants (Glycine max [L.] Merr. cv. Williams) was investigated at the vegetative stage (36 days after planting, DAP) and pod-filling stage (91 DAP) by the ¹⁵N pulse-chase experiment. The nodulated roots were exposed to N₂ gas labeled with a stable isotope ¹⁵N for 1 h, followed by 0, 1, 3 and 7 h of exposure with normal air. Plant roots and shoots were separated into three sections (basal, middle and distal parts) with the same length of the main stem or primary root. Approximately 80 and 92% of fixed N was distributed in the basal part of the nodulated roots at the vegetative and pod-filling stages by the end of 1 h of ¹⁵N₂ exposure, respectively. In addition, about 90% of fixed ¹⁵N was retained in the nodules and 10% was exported to root and shoot after 1 h of ¹⁵N₂ exposure at 91 DAP. The percentage distribution of ¹⁵N in the nodules at the pod-filling stage decreased from 90% to 7% during the 7 h of the chase period, and increased in the roots (14%), stems (54%), leaves (12%), pods (10%) and seeds (4%). The ¹⁵N distribution was negligible in the distal root segment, suggesting that N fixation activity was negligible and recycling fixed N from the shoot to the roots was very low in the initially short time of the experiment.     

Nitrogen isotope fractionation during nitrogen transport from ectomycorrhizal fungi,Suillus granulatus,to the host plant,Pinus densiflora

The ¹⁵N natural abundance in Pinus densiflora Sieb. et Zucco that had been inoculated and not inoculated with ectomycorrhizal fungi (Suillus granulatus (L.:Fr.) O. Kuntze) was compared. The inoculated pine needles showed a lower δ¹⁵N value, while the uninoculated ones showed a higher δ¹⁵N value. Higher δ¹⁵N values in the mycelial mat of the ectomycorrhizal fungi compared to those of the inoculated pine needles were also observed. These facts indicate that nitrogen isotope fractionation occurred during the nitrogen transport from mycorrhizal fungi to the host plants.     

Priming effects of 15N-labelled ammonium nitrate on uptake of soil N by wheat (Triticum aestivum L.) under field conditions

Summary The uptake of labelled and unlabelled N by wheat was measured in a field experiment using ¹⁵N-labelled ammonium nitrate fertilizer. The dry matter yield and N yields were significantly increased with fertilizer N application compared to those from unfertilized soil. The uptake of applied N by wheat ranged between 25 and 34%. Fertilizer N application increased the uptake of unlabelled soil N which was attributed to a positive priming effect or added N interaction. The added N interaction observed by applying 20, 60, and 120 kg fertilizer N was 11.4, 19.1, and 27.9 kg, corresponding to 26, 44 and 64%, respectively of the N taken up from unfertilized soil. The A values did not alter with the increase in fertilizer N application. The observed added N interaction may have been the result of pool substitution whereby added labelled fertilizer N stood proxy for unlabelled soil N. A significant correlation coefficient (r=0.996^**) between the uptake of soil N and the dry matter yield showed that soil N was more important than fertilizer N in wheat production.     

Dynamics of 15N-labeled ammonium sulfate in various inorganic and organic soil fractions of wetland rice soils

Summary The dynamics of basally applied ¹⁵N-labeled ammonium sulfate in inorganic and organic soil fractions of five wetland rice soils of the Philippines was studied in a greenhouse experiment. Soil and plant samples were collected and analyzed for ¹⁵N at various growth stages. Exchangeable NH₄ ⁺ depletion continued after 40 days after transplanting (DAT) and corresponded with increased nitrogen uptake by rice plants. Part of the applied fertilizer was fixed by 2:1 clay minerals, especially in Maligaya silty clay loam, which contained beidellite as the dominant clay mineral. After the initial fixation, nonexchangeable ¹⁵N was released from 20 DAT in Maligaya silty clay loam, but fixation delayed fertilizer N uptake from the soil. Part of the applied N was immobilized into the organic fraction. In Guadalupe clay and Maligaya silty clay loam, immobilization increased with time while the three other soils showed significant release of fertilizer N from the organic fraction during crop growth. Most of the immobilized fertilizer N was recovered in the nondistillable acid soluble (alpha-amino acid + hydrolyzable unknown-N) fraction at crop maturity. Between 61% and 66% of applied N was recovered from the plant in four soils while 52% of fertilizer N was recovered from the plant in Maligaya silty loam. Only 20% – 30% of the total N uptake at maturity was derived from fertilizer N. Nmin (mineral N) content of the soil before transplanting significantly correlated with N uptake. Twenty-two to 34% of applied N was unaccounted for possibly due to denitrification and ammonia volatilization.     

Comparative effects of applying leguminous and non-leguminous green manures and inorganic N on biomass yield and nitrogen uptake in flooded rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The beneficial role of green manures in rice production is generally ascribed to their potential of supplying plant nutrients, particularly nitrogen (N). However, the mechanisms through which green manures enhance the crop productivity are poorly understood. Pot experiments were conducted using a ¹⁵N-tracer technique: (1) to compare the biomass production potential of sesbania (Sesbania aculeata Pers.) and maize (Zea mays L.) as green manuring crops for lowland rice and (2) to compare the effect of the two types of green manure and inorganic N on the dry matter accumulation and N uptake by two rice (Oryza sativa L.) cultivars, viz. IR-6 and Bas-370. Although maize produced three times higher shoot biomass compared with sesbania, the latter showed higher N concentration; and thus the total N yield was similar in the two types of plants. Applying the shoot material of the two plants to flooded rice significantly enhanced the dry matter yield and N uptake by the two rice cultivars, the positive effects generally being more pronounced with sesbania than with maize amendment. The difference in the growth-promoting potential of the two plant residues was related more to an increased uptake of the native soil N rather than to their direct role as a source of plant-available N. A positive added nitrogen interaction (ANI) was observed due to both plant residues, the effect was much more pronounced with the application of sesbania than with maize residues. In both rice cultivars, inorganic N also caused a substantial ANI, particularly at higher application rate. Losses from the applied N were 2–3 times lower from sesbania, compared with maize treatment. Green manuring with sesbania also caused much lower N losses than the inorganic N applied at equivalent or higher rates. The overall benefit of green manuring to rice plants was higher than inorganic N applied at comparable rates. The two rice cultivars differed in their response to green manuring, IR-6 generally being more responsive than Bas-370.     

Effects of 15N-labelled ammonium nitrate and urea on soil nitrogen during growth of wheat (Triticum aestivum L.) under field conditions

We studied the effects of ¹⁵N-labelled ammonium nitrate and urea on the yield and uptake of labelled and unlabelled N by wheat (Triticum aestivum L., cv. Mexi-Pak-65) in a field experiment. The dry matter and N yields were significantly increased with fertilizer N application compared to those from unfertilized soil. The wheat crop used 33.6–51.5 and 30.5–40.9% of the N from ammonium nitrate and urea, respectively. Splitting the fertilizer N application had a significant effect on the uptake of fertilizer N by the wheat. The fertilizer N uptake showed that ammonium nitrate was a more available source of N for wheat than urea. The effective use of fertilizer N (ratio of fertilizer N in grain to fertilizer N in whole plant) was statistically similar for the two N fertilizers. The application of fertilizer N increased the uptake of unlabelled soil N by wheat, a result attributed to a positive added N interaction, which varied according to the fertilizer N split; six split applications gave the highest added N interaction compared to a single application or two split applications for both fertilizers. Ammonium nitrate gave 90.5, 33.5, and 48.5% more added N interaction than urea with one, two, and six split N applications. A values were not significantly correlated with the added N interaction (r=0.557). The observed added N interaction may have been the result of pool substitution, whereby added labelled fertilizer N replaced unlabelled soil N.     

Soil inorganic nitrogen composition and plant functional type determine forage crops nitrogen uptake preference in the temperate cultivated grassland,Inner Mongolia

ABSTRACT

Plant nitrogen (N)-acquisition strategy affects soil N availability, community structure, and vegetation productivity. Cultivated grasslands are widely established to improve degraded pastures, but little information is available to evaluate the link between N uptake preference and forage crop biomass. Here an in-situ ¹⁵N labeling experiment was conducted in the four cultivated grasslands of Inner Mongolia, including two dicots (Medicago sativa and Brassica campestris) and two monocots (Bromus inermis and Leymus chinensis). Plant N uptake rate, shoot- and root biomass, and concentrations of soil inorganic-N and microbial biomass-N were measured. The results showed that the root/shoot ratios of the dicots were 2.6 to 16.4 fold those of the monocots. The shoot N concentrations of the dicots or legumes were 40.6% to 165% higher than those of the monocots or non-legumes. The four forage crops in the cultivated grassland preferred to uptake more NO₃^?-N than NH₄⁺-N regardless of growth stages, and the NH₄⁺/NO₃^? uptake ratios were significantly lower in the non-legumes than in the legumes (p < 0.05). Significant differences in the NH₄⁺-N rather than NO₃^?-N uptake rate were observed among the four forages, related to plant functional types and growth stages. The NH₄⁺ uptake rate in the perennial forages exponentially decreased with the increases in shoot-, root biomass, and root/shoot ratio. Also, the plant NH₄⁺/NO₃^? uptake ratio was positively correlated with soil NH₄⁺/NO₃^? ratio. Our results suggest that the major forage crops prefer to absorb soil NO₃^?-N, depending on soil inorganic N composition and belowground C allocation. The preferential uptake of NO₃^?-N by forages indicates that nitrate-N fertilizer could have a higher promotion on productivity than ammonium-N fertilizer in the semi-arid cultivated grassland.     

Involvement of Bradyrhizobium japonicum denitrification in symbiotic nitrogen fixation by soybean plants subjected to flooding

Denitrification by Bradyrhizobium japonicum bacteroids contributes to nitric oxide (NO) production within soybean nodules in response to flooding conditions. However, the physiological relevance of NO production by denitrification in B. japonicum-Glycine max symbiosis is still unclear. In this work, soybean plants were inoculated with B. japonicum strains lacking the nirK or norC genes which encode the copper-containing nitrite reductase and the c-type nitric oxide reductase enzymes, respectively. 14 days flooding increased nodule number of plants inoculated with the WT and norC strains, but not of plants inoculated with the nirK mutant. However, nodule dry weight was not affected by 14 days flooding regardless of the strain used for inoculation. Supporting this observation, individual nodule growth was significantly higher in plants inoculated with nirK than those inoculated with WT or norC after 14 days flooding. Nodule functioning was strongly inhibited by flooding since leghemoglobin content of the nodules induced by any of the strains was significantly decreased after 7 or 14 days flooding compared to control plants. However, this effect was more relevant in nodules of plants inoculated with the WT or norC mutant than in those inoculated with the nirK mutant. Nitrogen fixation was also estimated by analyzing nitrogen content derived from biological nitrogen fixation in shoots, using the ¹⁵N isotope dilution technique. By using this approach, we observed that the negative effect of 14 days flooding on nitrogen fixation was more pronounced in plants inoculated with the norC mutant. However, nitrogen fixation of plants inoculated with nirK showed the highest tolerance to 14 days flooding. These findings allowed us to demonstrate the previously proposed hypothesis which suggests that NO formed by copper-containing nitrite reductase in soybean nodules, in response to flooding, has a negative effect on nitrogenase activity. We propose that inoculation of soybeans with a B. japonicum nirK mutant, which does not produce NO from nitrate, increases the tolerance of symbiotic nitrogen fixation to flooding.     

Time-course of nitrogen fixation in two bambara groundnut (Vigna subterranea L. Verdc.) cultivars

TheA-value method, involving the application of a higher¹⁵N rate to a reference non-N₂-fixing plant, was used to assess the magnitude of N₂ fixation in two bambara groundnut cultivars at four growth stages [vegetative, 0–47 days after planting (DAP); early pod-filling, 47–99 DAP; mid-pod-filling, 99–120 DAP; physiological maturity, 120–148 DAP). The cultivars were Ex-Ada, a bunchy type, and CS-88-11, a slightly spreading type. They were grown on a loamy sand. Uninoculated Ex-Ada and CS-88-11 were used as reference plants to measure the N₂ fixed in the inoculated bambara groundnuts. In this greenhouse study, soil was the major source of N in bambara groundnuts during vegetative growth, and during this period it accounted for over 80% of the N accumulaed in the plants. However, N₂ fixation became the major source of plant N during reproductive growth. There were significant differences between the two cultivars in the ability to fix N₂, and at physiological maturity, almost 75% of the N in CS-88-11 was derived from the atmosphere compared to 55% in Ex-Ada. Also, the total N fixed in CS-88-11 at physiological maturity was almost double that in Ex-Ada. Our data indicate that the higher N₂ fixation in CS-88-11 was due to two factors, a higher intensity of N₂ fixation and a longer active period of N₂ fixation. The results also suggest that bambara groundnut genotypes could be selected for higher N₂ fixation in farining systems.