Rhizospheric NO interacts with the acquisition of reduced N sources by the roots of European beech (Fagus sylvatica L.)

The gas phase of the soil plays an important role in plant growth and development. We investigated the effect of rhizospheric NO as a signalling compound for N uptake of beech roots. Following exposure to NO, ammonium and glutamine uptake into roots were determined using ¹⁵N-labelling, and gene expression of selected transporters was analysed by quantitative real-time PCR. Uptake of both N sources increased significantly with elevated NO concentration. However, with one exception, this increase was not reflected in up-regulation of expression of the respective transporters.     

Competition for nitrogen sources between European beech (Fagus sylvatica) and sycamore maple (Acer pseudoplatanus) seedlings

To investigate the short‐term consequences of direct competition between beech and sycamore maple on root N uptake and N composition, mycorrhizal seedlings of both tree species were incubated for 4 days (i.e. beech only, sycamore maple only or both together) in an artificial nutrient solution with low N availability. On the fourth day, N uptake experiments were conducted to study the effects of competition on inorganic and organic N uptake. For this purpose, multiple N sources were applied with a single label. Furthermore, fine roots were sampled and analysed for total amino acids, soluble protein, total nitrogen, nitrate and ammonium content. Our results clearly show that both tree species were able to use inorganic and organic N sources. Uptake of inorganic and organic N by beech roots was negatively affected in the presence of the competing tree species. In contrast, the presence of beech stimulated inorganic N uptake by sycamore maple roots. Both the negative effect of sycamore maple on N uptake of beech and the positive effect of beech on N uptake of sycamore maple led to an increase in root soluble protein in beech, despite an overall decrease in total N concentration. Thus, beech compensated for the negative effects of the tree competitor on N uptake by incorporating less N into structural N components, but otherwise exhibited the same strategy as the competitor, namely, enhancing soluble protein levels in roots when grown under competition. It is speculated that enhanced enzyme activities of so far unknown nature are required in beech as a defence response to inter‐specific competition.     

Rapid decline in nitrate uptake and respiration with age in fine lateral roots of grape: implications for root efficiency and competitive effectiveness

Changes in function as an individual root ages has important implications for understanding resource acquisition, competitive ability and optimal lifespan. Both nitrate uptake and respiration rates of differently aged fine roots of grape (Vitis rupestris x V. riparia cv. 3309 C) were measured. The resulting data were then used to simulate nitrate uptake efficiency and nutrient depletion as a function of root age. Both nitrate uptake and root respiration declined remarkably quickly with increasing root age. The decline in both N uptake and root respiration corresponded with a strong decline in root N concentration, suggesting translocation of nitrogen out of the roots. For simulations where no nutrient depletion occurs at the root surface, daily uptake efficiency was maximal at root birth and lifetime nitrate uptake efficiency slowly increased as the roots aged. Simulations of growth of roots into unoccupied soil using a solute transport model indicated the advantage of high uptake capacity in new roots under competitive conditions where nitrate availability is very transitory.     

Transformations in soil and availability to plants of15N applied as inorganic fertilizer and legume residues

Summary A pot experiment was conducted to study the transformations of organic and inorganic N in soil and its availability to maize plants. Inorganic N was in the form of¹⁵N labelled ammonium sulphate (As) and¹⁵N labelledSesbania aculeata (Sa), a legume, was used as organic N source. Plants utilized 20% of the N applied as As; presence of Sa reduced the uptake to 14%. Only 5% of the Sa-N was taken up by the plants and As had no effect on the availability of N from Sa. Losses of N from As were found to be 40% which were reduced to 20% in presence of Sa. Losses of N were also observed from Sa which increased in the presence of As. Application of As had no effect on the availability of soil or Sa-N. However, more As-N was transported into microbial biomass and humus components in the presence of Sa.Plants derived almost equal amounts of N from different sourcesi.e., soil, Sa and As. However, more As-N was transported into the shoots whereas the major portion of nitrogen in the roots was derived from Sa.     

Interactions in the uptake of amino acids, ammonium and nitrate ions in the Arctic salt-marsh grass, Puccinellia phryganodes

The uptake of amino acids and inorganic nitrogen by roots of Puccinellia phryganodes was examined to assess the potential contribution of soluble organic nitrogen to plant nitrogen uptake in Arctic coastal marshes, where free amino acids constitute a substantial fraction of the soil‐soluble N pool. Short‐term excised root uptake experiments were performed using tillers grown hydroponically under controlled conditions in the field. The percentage reductions in ammonium uptake at moderate salinity (150 mm NaCl) compared with uptake at low salinity (50 mm NaCl) were double those of glycine, but glycine uptake was more adversely affected than ammonium uptake by low temperatures. Glycine uptake was higher at pH 5·7 than at pH 7·0 or 8·2. The glycine uptake was up‐regulated in response to glycine, whereas ammonium uptake was up‐regulated in response to ammonium starvation. Nitrate uptake was strongly down‐regulated when tillers were grown on either ammonium or glycine. In contrast to N‐starved roots, which absorbed ammonium ions more rapidly than glycine, the roots grown on glycine, ammonium and nitrate and not N‐starved prior to uptake absorbed glycine as rapidly as ammonium and nitrate ions combined. Overall, the results indicate that amino acids are probably an important source of nitrogen for P. phryganodes in Arctic coastal marshes.     

13C AND 15N UPTAKE BY MARINE PHYTOPLANKTON. I. INFLUENCE OF NITROGEN SOURCE AND CONCENTRATION IN LABORATORY CULTURES OF DIATOMS1

Two species of marine diatoms, Skeletonema costatum (Grev.) Cleve and Phaeodactylum tricornutum Bohlin were grown in batch and continuous cultures on four different nitrogen compounds (nitrate, nitrite, ammonium, urea). Carbon and nitrogen uptake were measured simultaneously with the stable isotopes ¹³C and ¹⁵N. Nitrogen uptake generally increased with N concentration in the medium, but no clear difference existed between the N sources. Carbon fixation was decreased for up to 5 h following the addition of the N compound. Nitrite generally had the greatest inhibitory effect on C uptake. Carbon-to-nitrogen uptake ratios decreased with increasing dissolved N concentration, becoming lower than one in nutrient-limited cultures. In contrast, batch cultures exhibited C:N uptake ratios greater than one. These effects are essentially short-term and differ from long-term influences of the N source on the cellular chemical composition.     

Influence of ammonium on fine root development and rhizosphere pH of Douglas-fir seedlings in sand

Ammonium sulphate is a major component of the air pollutants deposited on forests in the Netherlands. Different amounts of NH₄ ⁺ were added to Douglas-fir seedlings grown in tall containers of sand, to study the influence of high concentrations of NH₄ ⁺ in the soil on the development of fine roots and the effects of nitrogen uptake on rhizosphere pH. At the end of this eight-month experiment part of the ammonium appeared to have nitrified into nitrate. High doses of ammonium negatively affected root length and root length per unit of dry matter (specific root length). Although Douglas fir shows a preferential ammonium uptake in nutrient solutions the increases in the pH of the rhizosphere in this experiment indicate that nitrogen was mostly taken up as nitrate. When the ammonium concentration in the soil is low, it cannot be taken up readily because of its low mobility in soil. Shoot growth was stimulated by high availability of nitrogen. The possible effects of high doses of ammonium on long-term forest vitality are discussed.     

Waterlogging and fire impacts on nitrogen availability and utilization in a subtropical wet heathland (wallum)

Protein, amino acids and ammonium were the main forms of soluble soil nitrogen in the soil solution of a subtropical heathland (wallum). After fire, soil ammonium and nitrate increased 90- and 60-fold, respectively. Despite this increase in nitrate availability after fire, wallum species exhibited uniformly low nitrate reductase activities and low leaf and xylem nitrate. During waterlogging soil amino acids increased, particularly γ-aminobutyric acid (GABA) which accounted for over 50% of amino nitrogen. Non-mycorrhizal wallum species were significantly (P < 0.05) ¹⁵N-enriched (0.3–4.3‰) compared to species with mycorrhizal associations (ericoid-type, ecto-, va-mycorrhizal) which were strongly depleted in ¹⁵N (-6.3 to -1.8‰). Lignotubers and roots had δ¹⁵N signatures similar to that of the leaves of respective species. The exceptions were fine roots of ecto-, ecto/va-, and ericoid type mycorrhizal species which were enriched in ¹⁵N (0.1–2.4‰). The 5¹⁵N signatures of δ¹⁵N_{total soil N} and δ¹⁵N_{soil NH4+} were in the range 3.7–4.5‰, whereas δ¹⁵N_{soil NO3?} was significantly (P < 0.05) more enriched in ¹⁵N (9.2–9.8‰). It is proposed that there is discrimination against ¹⁵N during transfer of nitrogen from fungal to plant partner. Roots of selected species incorporated nitrogen sources in the order of preference: ammonium > glycine > nitrate. The exception were proteoid roots of Hakea (Proteaceae) which incorporated equal amounts of glycine and ammonium.     

The use of mean pool abundances to interpret 15N tracer experiments

A pulse dilution ¹⁵N technique was used in the field to determine the effect of the ammonium to nitrate ratio in a fertilizer application on the uptake of ammonium and nitrate by ryegrass and on gross rates of mineralization and nitrification. Two experiments were performed, corresponding approximately to the first and second cuts of grass. Where no substantial recent immobilization of inorganic nitrogen had occurred, mineralization was insensitive to the form of nitrogen applied, ranging from 2.1–2.6 kg N ha^-1 d^-1. The immobilization of ammonium increased as the proportion of ammonium in the application increased. In the second experiment there was evidence that high rates of immobilization in the first experiment were associated with high rates of mineralization in the second. The implication was that some nitrogen immobilized in the first experiment was re-mineralized during the second. Whether this was nitrogen taken up, stored in roots and released following defoliation was not clear. Nitrification rates in this soil were low (0.1–0.63 kg N ha^-1 d^-1), and as a result, varying the ratio of ammonium to nitrate applied markedly altered the relative uptake of ammonium and nitrate. In the first experiment, where temperatures were low, preferential uptake of ammonium occurred, but where >90% of the uptake was as ammonium, a reduction in yield and nitrogen uptake was observed. In the second experiment, where temperatures and growth rates were higher, the proportion of ammonium to nitrate taken up had no effect on yield or nitrogen uptake.     

The fate of spring applied fertilizer N during the autumn-winter period: comparison between winter-fallow and green manure cropped soil

A field experiment was carried out at a pilot plot that was cropped with oilseed rape, and then left partly fallow and partly cropped with a green manure (mustard) during the autumn after harvest of the oilseed rape. The rape residues were incorporated in the soil. Methods used to quantify the N fluxes from harvest until sowing of the next crop were (1) ¹⁵N balance method, (2) total mineral N analysis and (3) NO emission measurements. Losses of spring applied fertilizer N were negligible in cropped plots and minimal in fallow plots during the following autumn-winter period. Most of the plant-N residues was retained by the organic N pool of the upper 30-cm soil layer. The green manure contributed slightly to soil available N at sowing of the next crop. However, the incorporation of plant material resulted in a nitrate flux that was at risk of leaching on the fallow plots, and on the green manure plots after incorporation of the green manure. This nitrate was largely derived from soil organic N, not from unused fertilizer applied in spring or from immobilized fertilizer. The NO emissions from the green manure plots were significantly higher than emissions from the fallow plots. The plants had a stimulating effect on the NO emission. A relationship between the NO emission and the soil nitrate concentration could not be established. No emissions were measured after green manure incorporation due to the low temperatures at the pilot plot. However, a greenhouse experiment showed an increased emission after incorporation. The NO emissions seemed to be related with the soil ammonium concentration.     

Nitrogen Use Efficiency of Crop Plants: Physiological Constraints upon Nitrogen Absorption

Current global nitrogen fertilizer use has reached approximately one hundred billion kg per annum. In many agricultural systems, a very substantial portion of this applied nitrogen fertilizer is lost from soil to groundwaters, rivers and oceans. While soil physicochemical properties play a significant part in these losses, there are several characteristic features of plant nitrogen transporter function that facilitate N losses. Nitrate and ammonium efflux from roots result in a reduction of net nitrogen uptake. As external nitrate and ammonium concentrations, respectively, are increased, particularly into the range of concentrations that are typical of agricultural soils, elevated rates of nitrate and ammonium efflux result. The rapid down-regulation of high-affinity influx as plants become nitrogen replete further reduces the root's capacity to acquire external nitrogen; only nitrogen-starved roots absorb with both high capacity and high affinity. The results of studies using molecular biology methods demonstrate that genes encoding nitrate and ammonium transporters are rapidly down-regulated when nitrogen is resupplied to nitrogen-starved plants. Provision of ammonium to roots of plants actively absorbing nitrate imposes a block on nitrate uptake, the extent of which depends on the ammonium concentration, thus further reducing the efficient utilization of soil nitrate. During the daily variation of incoming light and during periods of low incident irradiation (i.e. heavy cloud cover) the expression levels of genes encoding nitrate and ammonium transporters, and rates of nitrate and ammonium uptake, are substantially reduced. Low temperatures reduce growth and nitrogen demand, and appear to discriminate against high-affinity nitrogen influx. In sum, these several factors conspire to limit rates of plant nitrogen uptake to values that are well below capacity. These characteristics of the plant's nitrogen uptake systems facilitate nitrogen losses from soils.     

Nitrate assimilatory properties of barley grown under long-term N limitation: Effects of local nitrate supply in split-root cultures

The effect of nitrate availability on characteristics of the nitrate assimilatory system was investigated in N-limited barley (Hordeum valgare L. cv. Golf), grown with the seminal root system split into initially equal-sized halves. The cultures were continuously supplied with nitrate-N at a relative addition rate (RA) of 0.09 day^?1, which resulted in relative growth rates (RG) that were ca 85% of those observed under surplus nitrate nutrition. The total N addition was divided between the subroots in ratios of 100:0, 80:20, 70:30, 60:40, and 50:50. For comparison, standard cultures were grown at RAs ranging from 0.03 to 0.18 day^?1. Initially, biomass and N partitioning to the subroots responded strongly and proportionally to the nitrate distribution ratio. After 12-14 days no further effect was observed. The V_max for net nitrate uptake and in vitro nitrate reductase (NR) activity were measured in acclimated plants, i.e., after > 14 days under a certain nitrate regime. In subroots fed from 20 to 100% of the total N addition, V_max for net nitrate uptake increased slightly, whereas NR activity was unaffected. Uptake and NR activities were insignificant in the 0%-subroot. Uneven nitrate supply to individual subroots had negligible effect on the whole-plant ability for nitrate uptake, and the relative V_max (unit N taken up per unit N in whole plant tissue and time) remained about 7-fold in excess of the demand set by growth. Balancing nitrate concentrations (the resulting external nitrate concentrations at a certain RA) generally ranged between 2 and 10 μM at growth-limiting RA, both when predicted from uptake kinetics and when actually measured. When comparing split root and standard cultures when acclimated, it appears that uptake and NR activities in roots respond more strongly to over-all nitrate availability than to nitrate availability to individual subroots.     

Nitrate transport and signalling

Physiological measurements of nitrate (NO(3)(-)) uptake by roots have defined two systems of high and low affinity uptake. In Arabidopsis, genes encoding both of these two uptake systems have been identified. Most is known about the high affinity transport system (HATS) and its regulation and yet measurements of soil NO(3)(-) show that it is more often available in the low affinity range above 1 mM concentration. Several different regulatory mechanisms have been identified for AtNRT2.1, one of the membrane transporters encoding HATS; these include feedback regulation of expression, a second component protein requirement for membrane targeting and phosphorylation, possibly leading to degradation of the protein. These various changes in the protein may be important for a second function in sensing NO(3)(-) availability at the surface of the root. Another transporter protein, AtNRT1.1 also has a role in NO(3)(-) sensing that, like AtNRT2.1, is independent of their transport function. From the range of concentrations present in the soil it is proposed that the NO(3)(-)-inducible part of HATS functions chiefly as a sensor for root NO(3)(-) availability. Two other key NO(3)(-) transport steps for efficient nitrogen use by crops, efflux across membranes and vacuolar storage and remobilization, are discussed. Genes encoding vacuolar transporters have been isolated and these are important for manipulating storage pools in crops, but the efflux system is yet to be identified. Consideration is given to how well our molecular and physiological knowledge can be integrated as well to some key questions and opportunities for the future.     

Amino acid uptake in deciduous and coniferous taiga ecosystems

We measured in situ uptake of amino acids and ammonium across deciduous and coniferous taiga forest ecosystems in interior Alaska to examine the idea that late successional (coniferous) forests rely more heavily on dissolved organic nitrogen (DON), than do early successional (deciduous) ecosystems. We traced ¹⁵N-NH₄⁺ and ¹³C-¹⁵N-amino acids from the soil solution into plant roots and soil pools over a 24 h period in stands of early successional willow and late successional black spruce. Late successional soils have much higher concentrations of amino acid in soil solution and a greater ratio of DON to dissolved inorganic N (DIN) (ammonium plus nitrate) than do early successional soils. Moreover, late successional coniferous forests exhibit higher rates of soil proteolytic activity, but lower rates of inorganic N turnover. Differences in ammonium and amino acid uptake by early successional willow stands were insignificant. By contrast, the in situ uptake of amino acid by late successional black spruce forests were approximately 4-fold greater than ammonium uptake. The relative difference in uptake of ammonium and amino acids in these forests was approximately proportional to the relative difference of these N forms in the soil solution. Thus, we suggest that differences in uptake of different N forms across succession in these boreal forests largely reflect edaphic variation in available soil N (composition), rather than any apparent physiological specialization to absorb particular forms of N. These finding are relevant to our understanding of how taiga ecosystems may respond to increases in temperature, fire frequency, N deposition, and other potential consequences of global change.     

Forest fine-root production and nitrogen use under elevated CO2: contrasting responses in evergreen and deciduous trees explained by a common principle

Despite the importance of nitrogen (N) limitation of forest carbon (C) sequestration at rising atmospheric CO₂ concentration, the mechanisms responsible are not well understood. To elucidate the interactive effects of elevated CO₂ (eCO₂) and soil N availability on forest productivity and C allocation, we hypothesized that (1) trees maximize fitness by allocating N and C to maximize their net growth and (2) that N uptake is controlled by soil N availability and root exploration for soil N. We tested this model using data collected in Free‐Air CO₂ Enrichment sites dominated by evergreen (Pinus taeda; Duke Forest) and deciduous [Liquidambar styraciflua; Oak Ridge National Laboratory (ORNL)] trees. The model explained 80–95% of variation in productivity and N‐uptake data among eCO₂, N fertilization and control treatments over 6 years. The model explains why fine‐root production increased, and why N uptake increased despite reduced soil N availability under eCO₂ at ORNL and Duke. In agreement with observations at other sites, the model predicts that soil N availability reduced below a critical level diminishes all eCO₂ responses. At Duke, a negative feedback between reduced soil N availability and N uptake prevented progressive reduction in soil N availability at eCO₂. At ORNL, soil N availability progressively decreased because it did not trigger reductions in N uptake; N uptake was maintained at ORNL through a large increase in the production of fast turnover fine roots. This implies that species with fast root turnover could be more prone to progressive N limitation of carbon sequestration in woody biomass than species with slow root turnover, such as evergreens. However, longer term data are necessary for a thorough evaluation of this hypothesis. The success of the model suggests that the principle of maximization of net growth to control growth and allocation could serve as a basis for simplification and generalization of larger scale forest and ecosystem models, for example by removing the need to specify parameters for relative foliage/stem/root allocation.     

Effect of ammonium and nitrate application on the NO and N2O emission out of different soils

The effect of nitrate and ammonium application (0, 50, 100 and 150 mg N kg^-1 soil) was studied in an incubation experiment. Four Belgian soils, selected for different soil characteristics, were used. The application of both nitrate and ammonium caused an increase of the NO and N₂O emission. The NO production from nitrate and ammonium was found to be of the same order of magnitude. At low pH the NO production was found to be highest from nitrate, at higher pH values the production was found to be higher from ammonium. This seems to be the result of the negative effect of low pH on nitrification.The ANOVA analysis was carried out to separate the effect of the form of nitrogen, quantily of N applied and soil characteristics. The total production of NO was found to depend for 97% on the soil characteristics and for 3% on the quantity of N added. The total N₂O production depended for 100% on the soil characteristics.Stepwise regression analysis showed that the total NO production was best predicted by a combination of the factors CaCO₃ content and NH₄ ⁺ concentration in the soil. Total N₂O production was best described by a combination of CaCO₃, water soluble carbon (WSC) and sand-content.The N₂O/NO ratio was found to be highly variable, indicating that their productions react differently to changes in conditions, or are partly independent.It may be concluded that to NO and N₂O from soils both nitrification and denitrification may be equally important, their relative importance depending on local conditions such as substrate availability, water content of the soil etc. However, the NO production seems to be more nitrification dependent than the N₂O production. ei]{gnE}{fnMerckx}{edSection editor}     

Watershed liming effects on the forest floor N cycle

The forest floor was expected to play a major role in determining the total ecosystem response to watershed liming because of its high concentration of nutrients and its high level of activity. Net N mineralization and net nitrification were estimated in a field survey using the buried-bag approach. In a laboratory incubation experiment, forest floor humus was mixed with 6 doses of lime to determine the sensitivity of N mineralization and nitrification to lime dose. Forest floor microcosms with and without live tree roots were used to calculate a N budget for the system.The pH of the forest floor increased from 3.6 to 4.9 in the Oe and to 4.0 in the Oa two years after liming. The extractable ammonium pool in both the field survey and microcosm study was substantially smaller after liming and was probably a result of the 36% to 55% lower net N mineralization rate in limed plots than in reference plots. The laboratory incubation results agreed with the field survey results and further demonstrated that at higher lime doses (pH 5 to 6), N mineralization increased above controls. Net nitrification in limed humus in both the buried bags and laboratory incubation was as much as three times higher than controls, which could explain why nitrate leaching in limed microcosms was greater than in control microcosms. However, nitrate leaching from microcosms with live. roots was not affected by liming, suggesting that roots in the forest floor may prevent excess nitrate leaching. Reductions in N mineralization had no effect on N leaching or N uptake, but reduced the extractable ammonium pool.