15N-Labelled Ammonium and Nitrate Uptake by the Grass Calamagrostis villosa

Abstract

Calamagrostis villosa dominates the understory vegetation in declining spruce forests at higher elevations of the Central European mountain areas which show symptoms of needle yellowing and associated magnesium (Mg) deficiency. It was hypothesized that grasses would preferentially take up nitrate-nitrogen (NO₃-N) over ammonium-nitrogen (NH₄-N) which would support the cation balance in Mg deficient soils. In order to test this hypothesis, growth experiments were carried out in the greenhouse using plants which were cultivated in sand for nine weeks with full nutrient solution containing 0.2 or 2 mmol of N with different NH₄ ⁺ to NO₃ ^? ratios (1:0, 0.5:0.5, 0:1). In a short term experiment with labelled ¹⁵NH₄ ⁺ and ¹⁵NO₃ ^?, uptake of NH₄ ⁺ and NO₃ ^? was measured. When NO₃ ^? was the only N source it was taken up at similar rate per g dry mass as in the experiment in which NH₄ ⁺ was the only N source. However, at high supply pure NO₃ ^? nutrition resulted in higher biomass. In contrast, supply of only NH₄ ⁺ caused accumulation of N in the roots but growth remained restricted. If NH₄ ⁺ and NO₃ ^? were supplied at equal amounts, NH₄ ⁺ was the preferred form for N uptake. Biomass of the plants with mixed supply did not differ from the plants with pure NO₃ ^? nutrition.

The results point to an interesting interaction of carbon and nitrogen relation, but they do not support the initial hypothesis that grasses may prefer NO₃ ^? over NH₄ ⁺.     

The use of stable isotopes in ecosystem research. First results of a field study with 15N

Terrestrial ecosystems, e.g. forest ecosystems, are characterized by a complex and sensitive network of biotic and abiotic factors and their interactions. By using stable isotopes (e.g. labelled nitrogen compounds), very small addition rates of highly enriched compounds can be applied, which do not change or disturb the investigated system, but provide information about single processes, their interactions and especially about their dynamics.

First results of a field study in the Fichtelgebirge, Northeast-Bavaria, Germany, are presented. The distribution of labelled nitrogen (as ¹⁵N-NH₄ ⁺ and ¹⁵N[sbnd]NO₃ ^?) within a spruce ecosystem (Picea abies (L.) Karst. in competition with understory vegetation of Vaccinium myrtillus, Calluna vulgaris and Deschampsia flexuosa) showed maximum ¹⁵N concentrations in tissues of the understory vegetation. During the first six weeks after the ¹⁵N application, the nitrogen uptake of all investigated species was higher after the ¹⁵N[sbnd]NO₃ ^? treatment than after the ¹⁵N[sbnd]NH₄ ⁺ treatment.     

Distribution of Externally Applied 15NH4 + or 15NO3 − in White Lupins at Different Developmental Stages

White lupins (Lupinus albus L., var. Kievsky mutant) were grown in Mitscherlich pots. ¹⁵NH₄NO₃ or NH₄ ¹⁵NO₃ (10 or 95 at% ¹⁵N exc.) was applied at the late vegetative stage (14–16 leaves) or at the generative stage (end of bloom), respectively. Dry weight increase (ΔDW) of the plants as well as the ¹⁵N distribution in different organs and N fractions (NO_{3 ?}-N, NH₂-N, protein N) were investigated after 5 days and 50 days (maturity). The following results were obtained:

1. In comparison with the ¹⁵N derived from NO₃ ^?, the ¹⁵N derived from NH₄ ⁺ was more strongly retained in the roots. When transported into the shoot, it was more strongly translocated into the tip.

2. More ¹⁵N derived from NH⁴ ₊ was incorporated into proteins and reduced soluble N compounds than ¹⁵N derived from NO₄ ^?. Even in the above-ground plant tissues, a considerable proportion of the latter was found as NO₃ ^?.

3. In the long-term experiment (harvest at maturity 50 days after application), no difference in translocation and incorporation could be found between the two offered ¹⁵N sources.     

The Influence of Ammonium on Nitrate Uptake and Assimilation in 2-Year-Old Ash and Oak Trees - A Tracer-Study with 15N

Abstract

Interactions between ammonium and nitrate as competitive N sources depend on various biotic and abiotic factors. The preference for one of these N sources and the influence of ammonium on nitrate uptake and nitrate reductase activity was investigated in a ¹⁵N labelling experiment using 2-year-old potted plants of ash (Fraxinus excelsior L.) and oak (Quercus robur L.) under greenhouse conditions.

Seedlings of both tree species use ammonium and nitrate in equal amounts when both N forms are supplied in a 1:1 ratio (1.5 mM NH₄ ⁺ + 1.5 mM NO₃ ^?), although there is a slight tendency that ammonium is preferred. In both species total N uptake is higher if ammonium and nitrate are supplied simultaneously when compared with uptake of nitrate alone (3 mM nitrate). If nitrate is the sole N source N uptake is only half as high as if ammonium and nitrate are supplied in a ratio of 1:1.

The distribution of nitrate reductase between shoot and roots is not influenced by the N-form: nitrate reductase activity is always highest in the roots of both species under the conditions of this experiment.

Xylem sap analyses showed that both species transport higher concentrations of amino acids than of nitrate from the roots to the shoot. The amino acid composition is independent of the type of N source. Furthermore, ash trees contain more nitrate in the xylem sap than oak trees, reflecting the higher N uptake and the higher nitrate reductase activity in the leaves of this species.     

Synthese und Qualitätskontrolle 15N-markierter Stickoxide mit hoher 15N-Häufigkeit für umweltrelevante Tracer-Untersuchungen

Für die Untersuchung ausgewählter Probleme des Verhaltens und der Wirkung der Stickoxide NO _x (NO + NO₂) in Ökosystemen, z.B. die Aufnahme und Freisetzung von NOx durch das System “Boden-Pflanze”, bietet sich der Einsatz ¹⁵N-markierter Stickoxide an. Die dazu benötigten ¹⁵N-markierten Gasgemische hoher Reinheit werden aus eigens dafür synthetisiertem [¹⁵N]Stickstoffmonoxid oder [¹⁵N]Stickstoffdioxid mit hoher ¹⁵N-Häufigkeit hergestellt. Beide Synthesen gehen jeweils von der kostengünstig kommerziell erhältlichen [¹⁵N]Salpetersäure aus.

Im Falle des [¹⁵N]Stickstoffdioxids erfolgt die Herstellung über die Präparation von Bleinitrat und dessen thermische Zersetzung. Die Ausbeute liegt bei 70–75% bezogen auf eingesetzte [¹⁵N]Salpetersäure.

Die Herstellung von [¹⁵N]Stickstoffmonoxid erfolgt durch Reduktion von [¹⁵N]Salpetersäure mit Eisen-II-sulfat in stark saurer Lösung. Die Ausbeute beträgt 60–70%, bezogen auf eingesetzte Salpetersäure.

The application of ¹⁵N is very useful for the investigation of the behavior and the effect of the nitrogen oxides NO _x (nitric oxide + nitrogen dioxide) in ecosystems, e.g. the uptake and release of NO _x by the soil-plant system. The ¹⁵N labelled gas mixture needed for that purpose has to be prepared from synthesized highly enriched [¹⁵N]nitric oxide and [¹⁵N]nitrogen dioxide. These two syntheses both use the commercially available and reasonable [¹⁵N]nitric acid.

In the case of [¹⁵N]nitrogen dioxide the synthesis is carried out via [¹⁵N]lead nitrate and its decomposition with increasing temperature. The yield is 70–75% related to the [¹⁵N]nitric acid input. The preparation of [¹⁵N]nitric oxide is done by reduction of [¹⁵N]nitric acid by means of FeSO₄ in strong acid solution. The yield amounts to 60–70%.     

A Dual 13C and 15N Long Term Labelling Technique to Investigate Uptake and Translocation of C and N in Beech (Fagus sylvatica L.)

Abstract

A continuous dual ¹³CO₂ and ¹⁵NH₄ ¹⁵NO₃ labelling experimental set-up is presented that was used to investigate the C and N uptake and allocation within 3-year old beech (Fagus sylvatica L.) during one growing season. The C and N allocation pattern was determined after six, twelve and eighteen weeks of growth. The carbon uptake was distinctly different in the three phases examined: The first six weeks after budbreak were dedicated to leaf growth with a R/S (root to shoot) ratio of 0.14 for the new carbon. The second growth phase showed a balanced R/S ratio of C allocation and after week 13, the root compartment was the main carbon sink (R/S = 6.97).

Nitrogen allocation was more basipetal as compared to carbon. In the second growth phase, R/S of N_new was 5.57 but fell to 3.54 for the third growth phase probably due to formation of reserves in buds and stem.     

Incorporation of 15NO2 Nitrogen into Individual Amino Acids by Sunflowers Using Gc-C-Irms

Abstract

The nitrogen isotopic composition of individual amino acids in sunflower leaves after exposures to ¹⁵NO₂ in the range of ambient NO₂ concentrations (5–37 ppb) was analysed by Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry (GC-C-IRMS). Amino acids as well as the amides glutamine and asparagine were converted with MTBSTFA (N-methyl-N-(tert.-butyldimethylsilyl)-tri-fluoroacetamid) in pyridine to their corresponding TBDMS derivatives (N, O-tert.-butyldimethylsilyl) in a simple one-step silylation reaction. The derivatized amino acids were separated by gas chromatography, combusted on-line, and the products were sent continuously to an isotope ratio mass spectrometer. Accurate measurements were obtained, when more than 7 nmol N₂ were introduced into the ion source of the mass spectrometer per gas chromatographically separated and combusted compound. No interferences of the silicate and fluor containing derivatization agents on the performance of the system were observed.

In the range of ambient NO₂ concentrations sunflower leaves predominately incorporate the nitrogen derived from atmospheric NO₂ into soluble amino acids. The highest δ¹⁵N values were measured for alanine. The ¹⁵N enrichments of the detectable amino acids increased with increasing ¹⁵NO₂ concentration.     

Measuring of the Integral Airborne Nitrogen-Input into a Soil-Plant System by the 15N-Isotope Dilution Method

Abstract

Airborne nitrogen-inputs so far have only been measured in single fractions (deposition on plant surfaces or on soil and direct absorption of nitrogen containing gases by plants) by intensive exposition experiments in gas chambers, measurement of wet and dry N-deposition in the field and very expensive micrometereological field measurements. It is very difficult to estimate any overall N-input with practical relevance from these single N-component measurements. In this introduced field experiment an isolated measuring system is labelled with a ¹⁵N-tracer since it is not possible to label the nitrogen compounds of the atmosphere (e.g. NH₃, NO_x) which are to be absorbed. Through the dilution of this ¹⁵N-tracer by nitrogen derived from the atmosphere the total input of airborne nitrogen is determined. As soil resembling substrate sand was used and summer wheat was planted. With the regular and automated irrigation of nitrogen-free nutrient solution and the collection of precipitation surpluses this system measured the total input of airborne nitrogen for one vegetation period.

The first application of the described system gave an airborne nitrogen input into the soil-plant system during a vegetation period of 10 weeks (April-June) of about 32 kg ha^?1.     

The Relationship of Nitrate Reductase Activity to Uptake and Assimilation of Atmospheric 15NO2-Nitrogen in Needles of Norway Spruce (Picea abies [L.] Karst.)

Abstract

Using ¹⁵NO₂, relations between nitrate reductase activity and stomatal conductance, ¹⁵N-uptake and ¹⁵N-glutamate were studied in the two youngest needles flushes of potted Norway spruce (Picea abies [L.] Karst.). There were linear correlations between the stomatal conductance and the ¹⁵N-uptake and between the ¹⁵N-uptake and nitrate reductase (E.C. 1.6.6.1/1.6.6.2) activity. The ¹⁵N labelling of free glutamate shows the assimilation of NO₂ from the atmosphere in addition to the nitrogen from the soil. The portion of glutamate originating from ¹⁵NO₂ was linearly related to nitrate reductase activity in spring experiments. This indicates that this enzyme activity reflected the rate of NO₂-assimilation.     

Aufnahme von 15NO2 und Einbau des 15NO2-Stickstoffs in verschiedene Stickstofffraktionen bei Sonnenblumen (Uptake of 15NO2 and Metabolic Transfer of the 15NO2 Nitrogen to Various Nitrogen Fractions of Sunflowers)

Abstract

Sunflowers were exposed to ¹⁵NO₂ at a range between 4,9 and 42,0 ppb. The ¹⁵NO₂ uptake was quantified and the fate of the ¹⁵N in different parts and pools of nitrogen was investigated. The very high δ¹⁵N-values of the free amino acid pool can't give an answer to the question whether the NO₂-nitrogen is incorporated by a different pathway, compared to the nitrogen which derives from root uptake. Or does a compartmentation of the nitrate pool in the plant cell cause a dilution of the ¹⁵N enrichment in the mineral nitrogen pool during sample preparation?     

大气等离子体中氮氧化物粒子行为的数值模拟

利用一个空间零维大气等离子体模型对其中的氮氧化物在不同电离度情况下的变化规律进行了数值模拟,得到了放电后不同初始电子密度下的氮氧化物(包括NO,NO⁺,NO₂,NO₂⁺,N₂O,N₂O⁺,NO₃和N₂O₅)及影响其产消的主要反应物N和O₃的密度随时间的演化规律.结果表明,电子初始密度n_e0=10⁹ cm^-3时,NO和NO₂的去除率较高,氮氧化物总密度较小,最适合消除氮氧化物污染.同时,还对N和O₃随电离度变化的行为进行了分析. 关键词： 大气等离子体 氮氧化物 电离度 数值模拟     

Dinitrogen Fixation of Microbe-Plant Associations as Affected by Nitrate and Ammonium Supply

Abstract

The dinitrogen fixation activity of Azospirillum sp., and Pantoea agglomerans strains was determined by ¹⁵N₂ incorporation after incubation with ¹⁵N₂ labeled air or/and by acetylene reduction. These bacterial strains were able to fix N₂ both in pure culture and in association with wheat plants in hydroponics. Nitrogenase activity of Azospirillum sp., in pure culture was more rapidly inhibited by the addition of NH₄ ⁺ than NO₃ ^?. The N₂ fixation of P. agglomerans decreased only by NH₄ ⁺ -addition, but was stimulated by NO₃ ^?. Nitrogen fixation in association with wheat plants remained unaffected by both N compounds. However, nitrogen derived from the atmosphere (N_dfa) contributed only very little to the overall nitrogen nutrition of the plants.     

Inkorporation von 15N-markiertem Harnstoff in Organogene Dünge- und Abfallstoffe während der Rotte

Abstract

Eight organic fertilizers and wastes enriched with ¹⁵N labelled urea were stored at 25°C for one year. At the end of the experiment the ¹⁵N recovery rate ranged between 24 and 100%. The distributions of inorganic nitrogen (NH₄- and NO₃-N) and organic nitrogen (fulvic acids, humic acids and non extractable substances) are ascertained. Between the test materials, there are great differences in incorporation parameters.     

Lysimeter Investigations on the Effect of Winter Catch Crops and Weeded Fallow on the N-Dynamics in a Sandy Treposol Soil of Northeast Germany

Abstract

Lysimeter experiments (soil: sandy treposol, from the region “Havelländisches Luch”, Brandenburg, Germany) with application of ¹⁵N labelled fertilizer (80 kg N per ha as ¹⁵NH₄ ¹⁵NO₃, 10 at.-%¹⁵N exc.; for simulating mineralization in the early autumn period) were carried out to determine to what extent the amount of mineral- N was temporary conserved by winter catch crops, taken up subsequently in the vegetation periods by following crops, taken by subsequently in the vegetation periods by following crops, or percolated in the leaching water, respectively. The results were as follows:

1) Until winter or spring respectively, the catch crop uptake rates of applied mineral-N were 32% for phacelia (Phacelia tanacetifolia BENTH.), 25% for winter rape (Brassica napus L. cv. ‘AKELA’), and 16% for white mustard (Sinapis alba L.).

2) In the year after, following maize incorporated from 2.1 to 4.5% of the fertilizerborne N. The following plant community of fallow took up from 0.2 to 0.5% N originating from the fertilizer-N.

3) In comparison with the catch crops, N-leaching losses under fallow conditions were highest and equivalent to 17% of the applied fertilizer-N amount. In contrast to 3% of white mustard, phacelia and winter rape reduced N-leaching losses to 0.2 and 0.3% of the applied fertilizer-N amount.

4) In spring of the first year after the beginning of investigations, N-leaching losses were highest under fallow conditions and white mustard cultivation. Thus, the amounts of nitrate losses would exceed the EU limit for drinking water.

5) Three years after the investigations had been started, 10% (white mustard) and 20% (fallow) of the applied fertilizer-N was still found in th lysimeter soil.     

Using the natural 15N abundance to assess the main nitrogen inputs into the sand dune area of the North-Western Negev desert (ISRAEL)

The variation of the natural ¹⁵N abundance is often used to evaluate the origin of nitrogen or the pathways of N input into ecosystems. We tried to use this approach to assess the main input pathways of nitrogen into the sand dune area of the north-western Negev Desert (Israel). The following two pathways are the main sources for nitrogen input into the system:

1. Biological fixation of atmospheric nitrogen by cyanobacteria present in biological crusts and by N₂-fixing vascular plants (e.g. the shrub Retama raetam);

2. Atmospheric input of nitrogen by wet deposition with rainfall, dry deposition of dust containing N compounds, and gaseous deposition.

Samples were taken from selected environmental compartments such as biological crusts, sand underneath these crusts (down to a depth of 90?cm), N₂-fixing and non-N₂-fixing plants, atmospheric bulk deposition as well as soil from arable land north of the sandy area in three field campaigns in March 1998, 1999 and 2000. The δ¹⁵N values measured were in the following ranges: grass ?2.5‰ to +1.5‰; R. reatam: +0.5‰ to +4.5‰; non-N₂-fixing shrubs +1‰ to +7‰; sand beneath the biological crusts +4‰ to +20‰ (soil depth 2–90?cm); and arable land to the north up to 10‰. Thus, the natural ¹⁵N abundance of the different N pools varies significantly. Accordingly, it should be feasible to assess different input pathways from the various ¹⁵N abundances of nitrogen. For example, the biological N fixation rates of the Fabaceae shrub R. reatam from the ¹⁵N abundances measured were calculated to be 46–86% of biomass N derived from the atmosphere. The biological crusts themselves generally show slight negative ¹⁵N values (?3‰ to ?0.5‰), which can be explained by biological N fixation. However, areas with a high share of lichens, which are unable to fix atmospheric nitrogen, show very negative values down to ?10‰. The atmospheric N bulk deposition, which amounts to 1.9–3.8?kg?N/ha?yr, has a ¹⁵N abundance between 4.4‰ and 11.6‰ and is likely to be caused by dust from the arable land to the north. Thus, it cannot be responsible for the very negative values of lichens measured either. There must be an additional N input from the atmosphere with negative δ¹⁵N values, e.g. gaseous N forms (NO _x , NH₃). To explain these conflicting findings, detailed information is still needed on the wet, particulate and gaseous atmospheric deposition of nitrogen.     

Split-root labelling to investigate 15N rhizodeposition by Pinus sylvestris and Picea abies*

We investigated the transfer of ¹⁵N into the soil via ¹⁵N uptake and release by tree roots, which involves the principles of the split-root technique. One half of the root system received an injection of (¹⁵NH₄)₂SO₄ and the other half equivalent amounts of (NH₄)₂SO₄ at ¹⁵N natural abundance level. ¹⁵N was transferred from one side of the root system (¹⁵N side) to the other side (¹⁴N side) and released into the soil. The method was conducted with Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies [L.] Karst). Two concentration levels of (NH₄)₂SO₄ were used, corresponding with annual N deposition in the Netherlands (30 kg N ha^–1) and a twelfth of that (2.5 kg N ha^?1). Samples were taken 3 and 6 weeks after labelling and divided into needles?+?stem, roots, rhizosphere and bulk soil. Already 3 weeks after labelling, Scots pine took up 23.7?% of the low and 9.1?% of the high amounts of ¹⁵N, while Norway spruce took up 21.5 and 32.1?%, respectively. Both species transported proportions of ¹⁵N to the rhizosphere (0.1–0.2?%) and bulk soil (0.3–0.9?%). The method is a useful tool to investigate the fate of root-derived N in soils, for example, for the formation of stable forms of soil organic matter.     

15N Distribution in Wheat and Chemical Fractionation of Root-Borne 15N in the Soil

Abstract

Spring wheat plants were grown in split-root containers and labelled with ¹⁵N by fertilizing one compartment of the container with ¹⁵NH₄ ¹⁵NO₃ (95 at.-% ¹⁵N exc.). After the harvest, approx. 90% of the ¹⁵N incorporated by the plants were found in the shoots and 3% in the roots; approx. 7% had been released into the soil of the unlabelled compartment. The ¹⁵N in the soil of the unlabelled compartment was extracted with KCl and hydrolysed with HCl. Approx. 60% of the ¹⁵N was found in the hydrolysable organic N pool of the soil and 9% in the fraction of the soluble and exchangeable inorganic nitrogen.     

Laser Induced Energy Transfer Studies in Polyatomic Mixtures: CH3F-NO2

Abstract

The internal kinetics of NO₂ as well as the kinetics of crossover between CH₃F and NO₃ were studied in mixtures of these polyatomics and rare gases.

Subsequent to excitation of the CH₃F to v ₃=1 by the P(20) line of a 9.6μ Q-switch CO₂ laser, fluorescence of NO₂ at 6.12μ from v ₃=1 was monitored.

The equilibration rate of the stretching modes in NO₂ was determined to be 46 ± 10 msec^?1 torr^?1 while the rate of stretches-bend equilibration was measured as 15 ± 3 msec^?1 torr^?1. The rate of crossover from excited CH₃F to NO₂ in high rare gas dilution was measured as 90 ± 20 msec^?1 torr^?1 and the reverse rate as 285 ± 60 msec^?1 torr^?1.

Comparison to other triatomic systems as well as other experiments with mixtures of polyatomics are made and implications of laser “photochemistry” of NO₂ reactions are discussed.     

The Fate of [15N]Ammonium and [15N]Nitrate in the Soil of a 140-Year-Old Spruce Stand (Picea Abies) in the Fichtelgebirge (NE-Bavaria)

Abstract

A ¹⁵N tracer-experiment was carried out in a 140-year-old spruce stand (Picea abies (L.) Karst.) in the Fichtelgebirge (NE-Bavaria, Germany). Highly enriched (98 at%) [¹⁵N]ammonium and [¹⁵N]nitrate were applied as tracers by simulation of a deposition of 41.3 mol N ha^?1 with 11 water m^?2. To examine seasonal variations of uptake by spruce and understorey vegetation, different plots were labelled in spring, summer and autumn 1994.

One aim of the present study was to perfect a method of preparation of soil extracts for isotope ratio mass spectrometry (IRMS) measurements. Ammonium and nitrate from soil extracts were prepared for IRMS measurements by steam distillation and subsequent freeze drying. Additionally, tracer distribution and transformations in the soil nitrogen pools were examined. Ammonium, nitrate and total nitrogen were examined in the organic layer and the upper 10 cm of the mineral soil during 3 months after the first tracer application in spring 1994.

In July 1994, three months after tracer application, 40% of the [¹⁵N]ammonium label and 29% of the [¹⁵N]nitrate label, respectively, were recovered in the total N pool of the investigated soil horizons. In the organic layer the L/Of horizon retained most of the recovered tracers. Nitrification, immobilisation and mineralisation occurred even under the conditions of high soil acidity at the study site.     

Airborne Nitrogen Input at Four Locations in the German State of Saxony-Anhalt - Measurements using the 15N-Based ITNI-System

Abstract

The amount of atmospheric N deposition in Germany is actual rather uncertain. Estimates using standard methods indicate an N deposition of 30–35 kg N/ha × year. However, the results of long-term field experiments and newly by the ITNI (Integrated Total Nitrogen Input) system could prove a much higher N input of about 50–60 kg N/ha × year. The reason for this difference is that standard methods use wet-only or bulk collectors, which neglect gaseous and organic N deposition as well as direct N uptake by aerial plant parts. By contrast, the ITNI-system is able to measure the total atmospheric N input using the ¹⁵N isotope dilution method. The input of airborne N into a soil/plant system leads to a dilution of the abundance of a previously applied ¹⁵N tracer over a defined time period. The atmospheric N deposition can be calculated from this dilution.

To estimate the actual N input in Central Germany, ITNI measurements were carried out from autumn 1998 to autumn 2000 at four locations in the German state of Saxony-Anhalt. Atmospheric N depositions between 45 and 75 kg N/ha × year were determined depending on the location. These results closely match to N balances of longterm field experiments. Furthermore, a relationship was found between N deposition and the plant species used as well as plant development.