Assessing post-anthesis nitrogen uptake, distribution and utilisation in grain protein synthesis in barley (Hordeum vulgare L.) using 15N fertiliser and 15N proteinogenic and non-proteinogenic amino acids

We investigated the effects of nitrogen (N) availability during the vegetative phase on (a) post‐anthesis N uptake and (b) its translocation into ears in barley plants grown in a greenhouse at two levels of N: low (50 mg N kg^?1 sand) and optimal N supply (150 mg N kg^?1 sand). Plants in the two N treatments were fertilised with the same amount of labelled ¹⁵N [50 mg ¹⁵N kg^?1 sand at 10% ¹⁵N_exc (N_excess, i.e. N_exc, is defined as the abundance of enriched stable isotope minus the natural abundance of the isotope) applied as ¹⁵NH₄¹⁵NO₃] 10 days after anthesis (daa). In a separate experiment, the uptake and transport into ears of proteinogenic and non‐proteinogenic amino acids were studied to determine whether a relationship exists between amino acid transport into ears and their proteinogenic nature. Plants were fed with either ¹⁵N‐α‐alanine, a proteinogenic amino acid, or ¹⁵N‐α‐aminoisobutyric acid, a non‐proteinogenic amino acid. Both these amino acids were labelled at 95.6% ¹⁵N_exc. Results showed that N accumulations in stems, leaves and especially in ears were correlated with their dry matter (dm) weights. The application of 150 mg N kg^?1 sand significantly increased plant dm weight and total N accumulation in plants. During their filling period, ears absorbed N from both external (growth substrate) and internal (stored N in plants) sources. Nitrogen concentration in ears was higher in optimal N‐fed plants than in low N‐fed plants until 10 daa, but from 21 to 35 daa, differences were not detected. Conversely, ¹⁵N_exc in ears, leaves and stems was higher in low N‐fed plants than in optimal N‐fed plants. Ears acted as strong sink organ for the post‐anthesis N taken up from the soil independently of pre‐anthesis N nutrition: on average, 87% of the N taken up from the soil after anthesis was translocated and accumulated in ears. Low N‐fed plants continued to take up N from the post‐anthesis N fertiliser during the later grain‐filling period. The increase of pre‐anthesis N supply rate led to a decrease in the contribution of nitrogen derived from post‐anthesis ¹⁵N‐labelled fertiliser (N_dff) to total N in all aboveground organs, especially in ears where 44% and 22% of total N originated from post‐anthesis N uptake in low N‐fed and optimal N‐fed plants, respectively. The experiment with labelled amino acids showed that there was greater transport of proteinogenic amino acid into the ear (50% of total ¹⁵N) than non‐proteinogenic amino acid (39%). However, this transport of the non‐proteinogenic amino acids into ear suggested that the transport of N compounds from source (leaves) to sink organs (ear) might not be intrinsically regulated by their ability to be incorporated into storage protein of ears.     

Nitrogen incorporation and remobilization in different shoot components of field-grown winter oilseed rape (Brassica napus L.) as affected by rate of nitrogen application and irrigation

The seasonal course of nitrogen uptake, incorporation and remobilization in different shoot components of winter oilseed rape (Brassica napus L.) was studied under field conditions including three rates of ¹⁵N labelled nitrogen application (0, 100 or 200 kg N ha^-1) and two irrigation treatments (rainfed or watered at a deficit of 20 mm). The total amount of irrigation water applied was 260 mm, split over 13 occasions in a 7-week-period ranging from 1 week before onset of flowering until 4 weeks after flowering.Nitrogen application and irrigation increased plant growth and nitrogen accumulation. Irrespective of N and irrigation treatment more than 50% of total shoot N was present in the stem when flowering started. At the end of flowering, pod walls were the main N store containing about 30–40% of shoot N. The quantities of N remobilized from stems and pod walls amounted in all treatments to about 70% of the N present in these organs at mid-flowering. At harvest, stem and pod walls each contained about 10% of total shoot N, the remaining 80% being incorporated into seeds. The main component contributing to the response of seed N accumulation to nitrogen application and irrigation was pods in axillary racemes. Up to 20 kg N ha^-1, corresponding to about 10% of final shoot N content, was lost from the plants by leaf drop.Irrigation increased the recovery at harvest of applied N from 30% to about 50%, while the level of N application did not affect the N recovery. ¹⁵N labelled (fertilizer derived) nitrogen constituted a greater proportion of the N content in old leaves than in young leaves and increased with age in the former, but not in the latter. Relative to soil N, fertilizer derived N also contributed more to the N content of vegetative than to that of reproductive shoot components. Small net changes in shoot N content after flowering reflected a balance between N import and export, leading to continuous dilution of ¹⁵N labelled N with unlabelled N.     

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

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

The uptake and distribution of 15N2 into the various organs of Typha latifolia L.

Direct evidence of heterotrophic dinitrogen fixation associated with the emergent aquatic angiosperm, Typha latifolia L., was obtained through the exposure of actively growing plants to ¹⁵N₂ gas for 7 days in a gas-tight exposure vessel. Highest enrichments of ¹⁵N were found in roots/rhizomes and leaf bases. Slight enrichments were also found in the leaves due to translocation from the roots, rhizomes and leaf bases. Total fixed ¹⁵N values were 71.8 μg for the plant and 49.1 μg for the soil.Plants growing in silica sand, which received a nutrient solution containing combined nitrogen, exhibited higher enrichments and fixed 86% more ¹⁵N after exposure to ¹⁵N₂ gas than plants which received a nutrient solution lacking combined nitrogen. It is hypothesized that the concentration of combined nitrogen added was insufficient to repress nitrogen fixation and resulted in an increase in nitrogen fixation by associated microorganisms.Propane was used to trace the loss and movement of gases from the ¹⁵N₂ vessel and between the upper leaf chamber and the lower root chamber. Gas was rapidly exchanged between the upper and lower chambers through the leaves and roots of T. latifolia. Further investigations showed that propane moved at a rate of 1223 μmol day⁻¹ from the leaves to the roots and 2652 μmol day⁻¹ from the roots to the leaves. These data demonstrated that gases diffuse rapidly through the plant body of T. latifolia.     

Nitrogen metabolism and seed composition as influenced by foliar boron application in soybean

The physiological effects of foliar boron application (FB) on nitrogen metabolism and seed composition have not been well established in soybean [(Glycine max(L.)Merr.)]. Therefore, the effect of FB on nitrogen metabolism and seed composition was investigated. Nitrate assimilation was evaluated by measuring nitrate reductase activity (NRA) and nitrogen fixation was evaluated by measuring nitrogenase activity and natural abundance of ¹⁵N/¹⁴N. NRA were significantly (P?≤?0.05) higher in plants that received FB than the control plants. Higher rate of FB (One application of four times of commercial rate) inhibited nitrogen fixation as measured by natural abundance of ¹⁵N/¹⁴N ratio, but increased NRA. The higher activities of NR and nitrogenase by FB were accompanied with a higher B concentration in leaves. The significant (P?<?0.0001) enrichment of ¹⁵N/¹⁴N, accompanied with a higher rate of FB, suggested a possible mechanism where nitrate assimilation may compensate for the decrease in nitrogen fixation. FB increased seed protein by 13.7% and oleic acid by 30.9% compared to the control plants. This alteration was accompanied by a higher B concentration in leaves and seed. The results suggest that FB affects nitrogen metabolism and alters seed compositions, especially protein and unsaturated fatty acids.     

Intra‐trophic isotopic discrimination of 15N/14N for amino acids in autotrophs: Implications for nitrogen dynamics in ecological studies

The differential discrimination of nitrogen isotopes (¹⁵N/¹⁴N) within amino acids in consumers and their diets has been routinely used to estimate organismal tropic position (TP). Analogous isotopic discrimination can occur within plants, particularly in organs lacking chloroplasts. Such discrimination likely arises from the catabolic deamination of amino acids, resulting in a numerical elevation of estimated TP, within newly synthesized biomass. To investigate this phenomenon, we examined the ¹⁵N/¹⁴N of amino acids (δ¹⁵N_AA) in spring leaves and flowers from eight deciduous and two annual plants. These plants were classified on the basis of their time of bloom, plants that bloomed when their leaves were absent (Type I) versus plants that bloomed while leaves were already present (Type II). Based on the δ¹⁵N_AA values from leaves, both plant types occupied comparable and ecologically realistic mean TPs (=1.0 ± 0.1, mean ± 1σ). However, the estimated TPs of flowers varied significantly (Type I: 2.2 ± 0.2; Type II: 1.0 ± 0.1). We hypothesize that these results can be interpreted by the following sequence of events: (1) Type I floral biomass is synthesized in absence of active photosynthesis; (2) the catabolic deamination of amino acids in particular, leaves behind ¹⁵N in the residual pool of amino acids; and (3) the incorporation of these ¹⁵N‐enriched amino acids within the biomass of Type I flowers results in the numerical elevation of the TPs. In contrast, the actively photosynthesizing Type II leaves energetically sustain the synthesis of Type II flower biomass, precluding any reliance on catabolic deamination of amino acids. Amino acids within Type II flowers are therefore isotopically comparable to the Type II leaves. These findings demonstrate the idiosyncratic nature of the δ¹⁵N_AA values within autotrophic organs and have implications for interpreting trophic hierarchies using primary producers and their consumers.     

The nitrogen supply from soils and insects during growth of the pitcher plants Nepenthes mirabilis, Cephalotus follicularis and Darlingtonia californica

This study investigated the nitrogen (N) acquisition from soil and insect capture during the growth of three species of pitcher plants, Nepenthes mirabilis, Cephalotus follicularis and Darlingtonia californica. ¹⁵N/¹⁴N natural abundance ratios (δ¹⁵N) of plants and pitchers of different age, non-carnivorous reference plants, and insect prey were used to estimate proportional contributions of insects to the N content of leaves and whole plants. Young Nepenthes leaves (phyllodes) carrying closed pitchers comprised major sinks for N and developed mainly from insect N captured elsewhere on the plant. Their δ¹⁵N values of up to 7.2‰ were higher than the average δ¹⁵N value of captured insects (mean δ¹⁵N value = 5.3‰). In leaves carrying old pitchers that are acting as a N source, the δ¹⁵N decreased to 3.0‰ indicating either an increasing contribution of soil N to those plant parts which in fact captured the insects or N gain from N₂ fixation by microorganisms which may exist in old pitchers. The δ¹⁵N value of N in water collected from old pitchers was 1.2‰ and contained free amino acids. The fraction of insect N in young and old pitchers and their associated leaves decreased from 1.0 to 0.3 mg g⁻¹. This fraction decreased further with the size of the investigated tiller. Nepenthes contained on average 61.5 ± 7.6% (mean ± SD, range 50–71%) insect N based on the N content of a whole tiller. In the absence of suitable non-carnivorous reference plants for Cephalotus, δ¹⁵N values were assessed across a developmental sequence from young plants lacking pitchers to large adults with up to 38 pitchers. The data indicated dependence on soil N until 4 pitchers had opened. Beyond that stage, plant size increased with the number of catching pitchers but the fraction of soil N remained high. Large Cephalotus plants were estimated to derive 26 ± 5.9% (mean ± SD of the three largest plants; range: 19–30%) of the N from insects. In Cephalotus we observed an increased δ¹⁵N value in sink versus source pitchers of about 1.2‰ on average. Source and sink pitchers of Darlingtonia had a similar δ¹⁵N value, but plant N in this species showed δ¹⁵N signals closer to that of insect N than in either Cephalotus or Nepenthes. Insect N contributed 76.4 ± 8.4% (range 57–90%) to total pitcher N content. The data suggest complex patterns of partitioning of insect and soil-derived N between source and sink regions in pitcher plants and possibly higher dependence on insect N than recorded elsewhere for Drosera species. Received: 14 April 1997 / Accepted: 18 August 1997     

Estimation of symbiotic dinitrogen fixation in alder forest by the method based on natural15N abundance

Annual N₂-fixation in virgin forest ecosystems has been measured using a¹⁵N natural abundance (¹⁵N) procedure. This method was compared to a¹⁵N labelled fertilizer isotopic dilution method. For young alders (5–6 years old), ¹⁵N of leaves gave results in good agreement with the isotopic dilution of fertilizer method. Since ¹⁵N variability was expected according to plant physiology, for alder trees, leaves were collected at various heights after the end of the growing season, and, to take account of isotopic variations coming from derived inputs, ¹⁵N of leaves of a large number of other plants in the same are were measured to give control values. Following this procedure, the ¹⁵N method gave reliable evaluation of the nitrogen supply, by through N₂-fixation, to alders, which were found to maintain high nitrogen fixing capacity in a sequence ranging from first stage of establishment of climactic formation. Moreover, the same method is reported to discriminate various origins ofAlnus glutinosa grown in natural conditions, possibly in relation to the genetic diversity of this species.     

Is photosynthetic acclimation to free-air CO<Subscript>2</Subscript> enrichment (FACE) related to a strong competition for the assimilatory power between carbon assimilation and nitrogen assimilation in rice leaf?

Net photosynthetic rate (P _N) of leaves grown under free-air CO₂ enriched condition (FACE, about 200 μmol mol⁻¹ above ambient air) was significantly lower than P _N of leaves grown at ambient CO₂ concentration (AC) when measured at CO₂ concentration of 580 μmol mol⁻¹. This difference was found in rice plants grown at normal nitrogen supply (25 g m⁻²; NN-plants) but not in plants grown at low nitrogen supply (15 g m⁻²; LN-plants). Namely, photosynthetic acclimation to FACE was observed in NN-plants but not in LN-plants. Different from the above results measured in a period of continuous sunny days, such photosynthetic acclimation occurred in NN-plants, however, it was also observed in LN-plants when P _N was measured before noon of the first sunny day after rain. Hence strong competition for the assimilatory power between nitrogen (N) and carbon (C) assimilations induced by an excessive N supply may lead to the photosynthetic acclimation to FACE in NN-plants. The hypothesis is supported by the following facts: FACE induced significant decrease in both apparent photosynthetic quantum yield (Φ_c) and ribulose-1,5-bisphosphate (RuBP) content in NN-plants but not in LN-plants.     

Experimental evidence for diel δ15N‐patterns in different tissues,xylem and phloem saps of castor bean (Ricinus communis L.)

Nitrogen isotope signatures in plants might give insights in the metabolism and allocation of nitrogen. To obtain a deeper understanding of the modifications of the nitrogen isotope signatures, we determined δ¹⁵N in transport saps and in different fractions of leaves, axes and roots during a diel course along the plant axis. The most significant diel variations were observed in xylem and phloem saps where δ¹⁵N was significantly higher during the day compared with during the night. However in xylem saps, this was observed only in the canopy, but not at the hypocotyl positions. In the canopy, δ¹⁵N was correlated fairly well between phloem and xylem saps. These variations in δ¹⁵N in transport saps can be attributed to nitrate reduction in leaves during the photoperiod as well as to ¹⁵N‐enriched glutamine acting as transport form of N. δ¹⁵N of the water soluble fraction of roots and leaves partially affected δ¹⁵N of phloem and xylems saps. δ¹⁵N patterns are likely the result of a complex set of interactions and N‐fluxes between plant organs. Furthermore, the natural nitrogen isotope abundance in plant tissue is not constant during the diel course – a fact that needs to be taken into account when sampling for isotopic studies.     

Misconceptions and practical problems in the use of 15N soil enrichment techniques for estimating N2 fixation

The ¹⁵N methods are potentially accurate for measuring N₂ fixation in plants. The only problem with those methods is, how to ensure that the ¹⁵N/¹⁴N ratio in the plant accurately reflects the integrated ¹⁵N/¹⁴N ratio (R) in soil which is variable in time and with soil depth. However, the consequences of using an inappropriate reference plant vary with the level of N₂ fixation and the conditions under which the study was made. For example, the errors introduced into the values of N₂ fixation are higher at low levels of fixation, and decrease with increasing rates of fixation. At very high N₂ fixation rates, the errors are often insignificant. Also, the magnitude of error is proportional to the rate of decline of the ¹⁵N/¹⁴N ratio with time. Since N₂ fixation in most plants would be expected to below 60%, the question of how to select a good reference plant is still pertinent. In this paper, we have discussed some of the criteria to adopt in selecting reference plants, e.g. how to ensure that the reference plant is not fixing N₂, is absorbing most of its N from the same zone as the fixing plant, and in the same pattern with time, etc. In addition, we have discussed ¹⁵N labelling materials and methods that are likely to minimize any errors even when the fixing and reference plants don't match well in certain important criteria. The use of slow release ¹⁵N fertilizer or ¹⁵N labelled plant materials results in slow changes in the ¹⁵N/¹⁴N ratio of soil, and is strongly recommended. Where ¹⁵N inorganic fertilizers are used, the application of the fertilizer in small splits at various intervals is recommended over a one-time application. The problem with the reference crop, which has sometimes discouraged potential users of the ¹⁵N methods, is surmountable, as discussed in this paper.     

The Effect of Reduced Light Intensity and Sub-optimal Potassium Supply on N2 Fixation and N Turnover in Rhizobium Infected Lucerne

The effect of low light intensity and suboptimal potassium supply on the fixation of molecular nitrogen by root nodules and the nitrogen turnover in the host plant was studied in Medicago sativa using ¹⁵N labelled molecular nitrogen. For the application of ¹⁵N₂ labelled gas a special box was used. Both low light intensity and a low potassium supply resulted in a substantial growth depression. In particular the protein content of tops, roots and nodules was lower in the plants of the low light intensity treatment as compared with the control plants. Decreasing potassium supply had a similar but less-pronounced effect on protein content. The low protein content was not a consequence of a lack of soluble amino nitrogen or NH₃, since these fractions were influenced to a lesser degree by the reduced light intensity and by the low potassium supply. This observation is supported by the data obtained with ¹⁵N. N₂ fixation and NH₃ assimilation were affected by both low light intensity and low K application to the same degree as the overall metabolism, thus showing no particular response to the treatments applied.     

干旱胁迫下胡杨实生幼苗氮素吸收分配与利用

胡杨(Populus euphratica)是塔里木河流域荒漠河岸林的建群种,水分和氮素是限制胡杨幼苗的存活及早期生长的主要因子。利用~(15)N同位素示踪技术分析水和氮素的交互作用对胡杨幼苗不同生长阶段氮素的吸收分配利用及幼苗生长的影响,进一步探究氮素对胡杨实生苗早期形态建成的作用及对干旱胁迫的缓解效应,以期提高幼苗的存活率。实验以一年生胡杨实生幼苗为研究对象,采用温室内盆栽实验,设置4个干旱处理(D_1、D_2、D_3、D_4,土壤相对含水量为:20%—25%、40%—45%、60%—65%、80%—85%)和3种氮素水平(N_0、N_1、N_2:0、3、6 g/盆)测定胡杨幼苗的生长指标和各部分的Ndff、分配率及利用率。结果表明:胡杨幼苗在土壤相对含水量60%—65%(D_3)、氮素添加量3 g/盆(N_1)时,其生长表现为最佳状态;干旱胁迫下,不同氮素添加量对胡杨幼苗各部分的Ndff值存在显著差异,N_2低于N_1;随干旱胁迫减弱(D_3、D_4),植株在生长早期(25 d)根部吸收的~(15)N优先向地上部分转运,生长后期(75 d)植株Ndff最高,其中以根系中Ndff最高;不同生长期幼苗各部分的~(15)N分配存在显著差异,根系~(15)N分配率较高,但不同氮量处理间差异不显著;随生长期的推移,植株对~(15)NH_4~(15)NO_3的利用率表现为粗根最大,各处理中D_3N_1处理均显著高于其他处理。结论:轻度干旱胁迫下添加适量的氮素能够增强植株对氮素的吸收征调能力,优化水资源获取以维持生存的重要机制。     

Bi-directional transfer of nitrogen between alfalfa and bromegrass: Short and long term evidence

Transfer of N from legumes to associated non-legumes has been demonstrated under a wide range of conditions. Because legumes are able to derive their N requirements from N₂ fixation, legumes can serve, through the transfer of N, as a source of N for accompanying non-legumes. Studies, therefore, are often limited to the transfer of N from the legume to the non-legume. However, legumes preferentially rely on available soil N as their source of N. To determine whether N can be transferred from a non-legume to a legume, two greenhouse experiments were conducted. In the short-term N-transfer experiment, a portion of the foliage of meadow bromegrass (Bromus riparius Rhem.) or alfalfa (Medicago sativa L.) was immersed in a highly labelled ¹⁵N-solution and following a 64 h incubation, the roots and leaves of the associated alfalfa and bromegrass were analyzed for ¹⁵N. In the long-term N transfer experiment, alfalfa and bromegrass were grown in an ¹⁵N-labelled nutrient solution and transplanted in pots with unlabelled bromegrass and alfalfa plants. Plants were harvested at 50 and 79 d after transplanting and analyzed for ¹⁵N content. Whether alfalfa or bromegrass were the donor plants in the short-term experiment, roots and leaves of all neighbouring alfalfa and bromegrass plants were enriched with ¹⁵N. Similarly, when alfalfa or bromegrass was labelled in the long-term experiment, the roots and shoots of neighbouring alfalfa and bromegrass plants became enriched with ¹⁵N. These two studies conclusively show that within a short period of time, N is transferred from both the N₂-fixing legume to the associated non-legume and also from the non-legume to the N₂-fixing legume. The occurrence of a bi-directional N transfer between N₂-fixing and non-N₂-fixing plants should be taken into consideration when the intensity of N cycling and the directional flow of N in pastures and natural ecosystems are investigated.     

模拟氮沉降和接种外生菌根真菌对马尾松(Pinus massoniana)幼苗营养元素的影响

量化植物地上部和地下部元素含量对于理解和预测植物养分平衡如何响应大气氮沉降的变化至关重要。通过盆栽试验研究了氮沉降增加背景下外生菌根真菌对马尾松幼苗营养元素的影响。对马尾松幼苗进行了接种两种外生菌根真菌：（彩色豆马勃（Pisolithus tinctorius,Pt）与厚环乳牛肝菌（Suillus grevillei,Sg））以及4种氮素浓度添加：0 kg N hm^-2a^-1（N0）、正常氮沉降30 kg N hm^-2a^-1（N30）、中度氮沉降60 kg N hm^-2a^-1（N60）、重度氮沉降90 kg N hm^-2a^-1（N90）,共12个处理,测定了马尾松地上部和地下部大量元素和微量元素的含量。结果表明：施氮改变了营养元素在马尾松幼苗地上部和地下部的含量,马尾松幼苗磷（P）、钙（Ca）、铁（Fe）、锰（Mn）等元素均在N60时达到临界值,而当输入的量超过了马尾松对氮的需求时,氮沉降会使马尾松营养元素含量较最适浓度时降低,地上部碳（C）随施氮浓度的升高先升高后降低,N随施氮浓度的升高而升高,根系和叶片钾（K）、Ca、镁（Mg）均随施氮浓度的升高而降低,施氮也降低了根系C及微量元素的含量。但在同一施氮浓度下,接种外生菌根真菌（EMF）后能够提高大多数元素的含量,N90时接种厚环乳牛肝菌（Sg）和彩色豆马勃（Pt）的叶片N含量与对照相比分别提高112.6%和138.6%,根系N含量分别提高73.1%、71.6%;N60时接种Sg和Pt的植株叶片P含量比不施氮未接种对照分别提高了166.3%、132.9%,根系P含量分别提高了40.8%、38.5%。EMF能够维持植物养分平衡,从而降低高施氮量对植物的影响效果。这为未来气候变化情景中氮沉降增加下接种EMF可以调节植物元素含量,从而达到更适应环境的元素平衡来促进生长提供理论依据。     

Alder and lupine enhance nitrogen cycling in a degraded forest soil in Northern Sweden

Positive effects of legumes and actinorhizal plants on N-poor soils have been observed in many studies but few have been done at high latitudes, which was the location of our study. We measured N₂ fixation and several indices of soil N at a site near the Arctic Circle in northern Sweden. More than 20 years ago lupine (Lupinus nootkatensis Donn) and gray alder (Alnus incana L. Moench) were planted on this degraded forest site. We measured total soil N, net N mineralization and nitrification with a buried bag technique, and fluxes of NH⁺ ₄ and NO^– ₃ as collected on ion exchange membranes. We also estimated N₂ fixation activity of the N₂-fixing plants by the natural abundance of ¹⁵N of leaves with Betula pendula Roth. as reference species. Foliar nitrogen in the N₂-fixing plants was almost totally derived from N₂ fixation. Plots containing N₂-fixing species generally had significantly higher soil N and N availability than a control plot without N₂-fixing plants. Taken together, all measurements indicated that N₂-fixing plants can be used to effectively improve soil fertility at high latitudes in northern Sweden.     

Distribution of fixed-N and nitrate-N in cowpea during pod development

If the quality and quantity of yields from cowpea (Vigna unguiculata [L.] Walp.) are to be maximised, a complete understanding of the N nutrition of the plant must be achieved. The N requirement for developing pods of this species may come from mobilization of N in vegetative tissue, biological N fixation and uptake of N from soil. In this study, the fate of a pulse of fixed ¹⁵N₂ or of ¹⁵NO₃-given to different cowpea plants during pod development was determined. The plants were grown in vermiculite in plastic pots that were able to be sealed with silicone adhesive and equipped with a rubber septum so that ¹⁵N₂ gas could be injected into the air space above the vermiculite, and gas losses would be eliminated. Nineteen days after injection of ¹⁵N₂ the pods, leaves, nodules and roots contained 65%, 15%, 9%, and 4%, respectively of the quantity of ¹⁵N₂ fixed. When ¹⁵NO₃-¹⁵N was taken up by other plants during this period, these plant parts contained 40%, 26%, 3% and 19%, respectively, of the total plant ¹⁵N. The percentage ¹⁵N in roots was greater, and that of ¹⁵N in nodules was lower, when ¹⁵NO₃-¹⁵N was applied than when ¹⁵N₂ was utilised by plants. These results indicate that, while a high percentage of fixed-N or NO₃-N given to cowpea plants moved to the developing pods, other sinks were competing for this newly-aquired N.     

Estimation of N2-fixation by sugarcane,Saccharum sp., and soybean,Glycine max,grown in soil with15N-labelled organic matter

Summary Two varieties of sugarcane, and nodulated and non-nodulated soybean isolines, were planted in a soil previously mixed with¹⁵N-labelled plant material. 45 days was allowed to elapse before planting, to permit initiation of organic matter mineralization. Plants were grown for 60 days, then harvested, dried, weighed and analysed for total N. Analysis of soil samples pre-incubated in the laboratory was carried out to evaluate ammonium and nitrate from added organic matter. Dry weights of the soybean isolines were similar, but total N was higher for the nodulated line. Both sugarcane varieties showed similar weight and total N. Nitrogen derived from applied organic matter (NdfOM) was higher in non-nodulated soybean than in all other plants. Although there is the possibility of different¹⁵N availabilities between species, nitrogen derived from fixation (Nfix) was calculated based on the¹⁵N enrichment of the non-nodulating soybean. Nfix was 72% for nodulating soybean and ranged from 19 to 39% for different parts of sugarcane plants, despite high levels of available-N. Nitrogen derived from soil was calculated by difference. NdfOM was lower in roots than in upper parts (leaves+stalks) of plants. Use of¹⁵N labelled organic matter seems a useful approach to the longer term measurement of N₂-fixation.IAEA Project BRA/5/009-CENA.     

Contribution of N2 fixing cyanobacteria to rice production: availability of nitrogen from 15N-labelled cyanobacteria and ammonium sulphate to rice

This study investigate the potential contribution of nitrogen fixation by indigenous cyanobacteria to rice production in the rice fields of Valencia (Spain). N₂-fixing cyanobacteria abundance and N₂ fixation decreased with increasing amounts of fertilizers. Grain yield increased with increasing amounts of fertilizers up to 70 kg N ha^-1. No further increase was observed with 140 kg N ha^-1. Soil N was the main source of N for rice, only 8–14% of the total N incorporated by plants derived from ¹⁵N fertilizer. Recovery of applied ¹⁵N-ammonium sulphate by the soil–plant system was lower than 50%. Losses were attributed to ammonia volatilization, since only 0.3–1% of applied N was lost by denitrification. Recovery of ¹⁵N from labeled cyanobacteria by the soil–plant system was higher than that from chemical fertilizers. Cyanobacterial N was available to rice plant even at the tillering stage, 20 days after N application. This revised version was published online in June 2006 with corrections to the Cover Date.     

Variation of 15N natural abundance in leaves and nodules of actinorhizal shrubs in Northwest Patagonia

This study was performed to assess the N₂-fixing capability of the native actinorhizal species Ochetophila trinervis (sin. Discaria trinervis) and Discaria chacaye (Rhamnaceae) in Northwest Patagonia. We measured the N concentration and ¹⁵N natural abundance in leaves and nodules of O. trinervis and D. chacaye, in leaves of associated non-actinorhizal vegetation, and in the soils under each sampled plant. O. trinervis and D. chacaye had foliar N concentrations that were about twice that of non-actinorhizal shrubs growing at the same sites, even though soils varied four-fold in total N across the sites. Leaves of both actinorhizal plants had a similar δ¹⁵N at any site and were close to atmospheric values. The foliar δ¹⁵N of non-actinorhizal plants and soil δ¹⁵N were strongly correlated across the sites. Nodules were depleted in δ¹⁵N relative to the foliage of the respective actinorhizal species. In conjunction with the uniformly high foliage N concentration of these actinorhizal plants and the universal presence of vesicles observed in root nodules, these data strongly suggest that O. trinervis and D. chacaye obtain a significant amount of their N from N₂ fixation. To calculate the proportion of N derived from atmosphere, theoretical B-values were estimated. In all cases where the δ¹⁵N of fixing and reference foliage were significantly different, O. trinervis and D. chacaye obtained almost all of their N from N₂ fixation. These results are the first to demonstrate N₂ fixation by O. trinervis and D. chacaye in the field and therefore suggest an important role for these actinorhizal plants in the N economy of ecosystems in northwest Patagonia as well as their potential use for restoration of degraded lands in this region.