Change in uptake, transport and accumulation of ions in Nerium oleander (rosebay) as affected by different nitrogen sources and salinity

Background and Aims

The source of nitrogen plays an important role in salt tolerance of plants. In this study, the effects of NaCl on net uptake, accumulation and transport of ions were investigated in Nerium oleander with ammonium or nitrate as the nitrogen source in order to analyse differences in uptake and cycling of ions within plants.

Methods

Plants were grown in a greenhouse in hydroponics under different salt treatments (control vs. 100 mm NaCl) with ammonium or nitrate as the nitrogen source, and changes in ion concentration in plants, xylem sap exuded from roots and stems, and phloem sap were determined.

Key Results

Plant weight, leaf area and photosynthetic rate showed a higher salt tolerance of nitrate-fed plants compared with that of ammonium-fed plants. The total amount of Na⁺ transported in the xylem in roots, accumulated in the shoot and retranslocated in the phloem of ammonium-fed plants under salt treatment was 1·8, 1·9 and 2·7 times more, respectively, than that of nitrate-treated plants. However, the amount of Na⁺ accumulated in roots in nitrate-fed plants was about 1·5 times higher than that in ammonium-fed plants. Similarly, Cl⁻ transport via the xylem to the shoot and its retranslocation via the phloem (Cl⁻ cycling) were far greater with ammonium treatment than with nitrate treatment under conditions of salinity. The uptake and accumulation of K⁺ in shoots decreased more due to salinity in ammonium-fed plants compared with nitrate-fed plants. In contrast, K⁺ cycling in shoots increased due to salinity, with higher rates in the ammonium-treated plants.

Conclusions

The faster growth of nitrate-fed plants under conditions of salinity was associated with a lower transport and accumulation of Na⁺ and Cl⁻ in the shoot, whereas in ammonium-fed plants accumulation and cycling of Na⁺ and Cl⁻ in shoots probably caused harmful effects and reduced growth of plants.Key words: Mineral cycling, Nerium oleander, nitrogen source, salinity, xylem and phloem transport     

The uptake and flow of C, N and ions between roots and shoots in Ricinus communis L. III. Long-distance transport of abscisic acid depending on nitrogen nutrition and salt stress

Seedlings of Ricinus communis L. were cultivated in quartz sandand supplied with media which contained either different concentrationsof nitrate or ammonium nitrogen and were treated with a lowsalt stress. The concentration of ABA was determined in tissuesand in xylem and phloem saps. Between 41 and 51 day after sowing,abscisic acid (ABA) flows between roots and shoots were modelled.Long-distance transport of ABA was not stimulated under conditionsof nitrate deficiency (0.2 mol m^–3). However, when ammoniumwas given as the only N source (1.0 mol m^–3), ABA transportin both xylem and phloem was increased significantly. Mild saltstress (40 mol m^–3 NaCl) increased ABA transport in nitrate-fedplants, but not in ammonium-fed plants. The leaf conductancewas lowered by salt treatment with both nitrogen sources, butit was always lower in ammonium-fed compared to nitrate-fedplants. A negative correlation of leaf conductance to ABA levelsin leaves or flow in xylem was found only in comparison of ammonium-fedto nitrate-fed plants. Key words: Abscisic acid, ammonium, Ricinus communis, phloem, xylem, transport, nitrate, nitrogen nutrition     

Morphophysiological Indices of the Source Leaf in Wheat Plants Acclimated to Conditions of Nitrogen Nutrition

Growth, leaf and cell morphology, and the chemical composition of the second leaf were studied in wheat (Triticum aestivumL., cv. Inna) plants grown on the medium containing nitrate, ammonium, or no nitrogen at all. Independent of the nitrogen nutrition, the second leaf of the 21-day-old plants matures and functions as a source of assimilates. Both ammonium nutrition and nitrogen deficiency decreased the fresh weight, area, and cell size in the leaf; however, the conditions of nitrogen nutrition did not affect the dry weight of the leaf. Nitrogen starvation increased and ammonium nutrition decreased the relative content of the cell walls in the dry weight. In the nitrate-fed plants, the leaf content of sucrose increased, and the contents of reduced nitrogen (N_red) and protein were lower than in the ammonium treatment. Reciprocally, the contents of reduced nitrogen and protein were highest in the ammonium treatment, the content of sucrose was lowest, with starch practically absent from the leaf. The nitrogen-starved leaf accumulated a large amount of starch, the N_redcontent was two times lower than in the ammonium-fed plants, and the protein content was similar to that in the nitrate-fed plants. Thus, leaf and cell morphology and the content of N_red, protein, and carbohydrate changes in different ways during wheat acclimation to the condition of nitrogen nutrition. By assessing the cell wall weight, the authors established that, depending on nitrogen nutrition, this cell compartment accepts a variable flow of carbon.     

Oxalate Accumulation as Regulated by Nitrogen Forms and Its Relationship to Photosynthesis in Rice (Oryza sativa L.)

Four-leaf rice seedlings (Oryza sativa L.), which had been cultivated in Kimura B complete nutrient solution, were treated with two nitrogen forms by replacing the nitrogen element in the complete solution with sole nitrate or ammonium (2.86 mmol/L). Nitrate-N nutrition tended to increase oxalate content in all parts of the plant, including the leaves, stems, roots, and root exudates, whereas ammonium had the opposite effect. Consequently, marked differences in oxalate content were observed between the two treatments throughout the time tested (0--12 d), with maximal differences of approximately 12-fold at 6 d after treatment. Photosynthetic/respiratory parameters were examined over time simultaneously with changes in oxalate content. Net photosynthetic rate, chlorophyll fluorescence parameters (i.e. maximal photochemical efficiency (Fv/Fm) and photochemical quantum yields of photosystem (PS)Ⅱ (φ PSⅡ)), and respiratory rate were not significantly different between plants treated with the two nitrogen forms, although ammonium-fed plants had apparently higher leaf chlorophyll content than nitrate-fed plants. Leaf glucose content was altered little, but the content of fructose, sucrose, and total soluble sugar was significantly higher in the leaves of ammonium-fed plants than nitrate-fed plants, The results indicate that nitrate/ammonium may serve as efficient regulators of oxalate accumulation owing to regulation of metabolism in rice leaves rather than oxalate downward transfer and root excretion, and that photosynthetic metabolism is not directly correlated with the regulation of oxalate accumulation in rice plants.     

Nitrogen uptake and metabolism in Populus x canescens as affected by salinity

External salinization can affect different steps of nitrogen (N) metabolism (ion uptake, N assimilation, and amino acid and protein synthesis) depending on the inorganic N source. Here, we assessed the net uptake of N supplied as nitrate or ammonium and N assimilation (combining metabolite analyses with molecular biological approaches) in grey poplar (Populus x canescens) plants grown under saline (75 mM NaCl) and control conditions. The specific (micromol N g(-1) dry weight fine roots h(-1)) and total plant (micromol N per plant h(-1)) N net uptake rates, total plant N content, total plant biomass and total leaf protein concentration were reduced under saline conditions when plants were supplied with ammonium. In both nutritional groups, salt treatment caused pronounced accumulation of soluble N compounds in the leaves. The mRNAs of genes coding for enzymes catalyzing rate-limiting steps of both proline synthesis and degradation (delta-1-pyrroline-5-carboxylate synthase and proline dehydrogenase) as well as for NADH-dependent glutamate synthase were accumulated under saline conditions. Whereas under control conditions the plant N status seemed to be superior when ammonium was supplied, the N balance of ammonium-fed plants was more severely affected by salt stress than that of plants supplied with nitrate. Possible metabolic implications of stress-related accumulation of particular amino acids are discussed.     

Oxalate Accumulation as Regulated by Nitrogen Forms and Its Relationship to Photosynthesis in Rice （Oryza sativa L.）

Four-leaf rice seedlings (Oryza sativa L.), which had been cultivated in Kimura B complete nutrient solution, were treated with two nitrogen forms by replacing the nitrogen element in the complete solution with sole nitrate or ammonium (2.86 mmol/L). Nitrate-N nutrition tended to increase oxalate content in all parts of the plant, including the leaves, stems, roots, and root exudates, whereas ammonium had the opposite effect. Consequently, marked differences in oxalate content were observed between the two treatments throughout the time tested (0-12 d), with maximal differences of approximately 12-fold at 6d after treatment. Photosynthetic/respiratory parameters were examined over time simultaneously with changes in oxalate content. Net photosynthetic rate, chlorophyll fluorescence parameters (i.e. maximal photochemical efficiency (Fv/Fm) and photochemical quantum yields of photosystem (PS)II (ΦPSⅡ)), and respiratory rate were not significantly different between plants treated with the two nitrogen forms, although ammonium-fed plants had apparently higher leaf chlorophyll content than nitrate-fed plants. Leaf glucose content was altered little, but the content of fructose, sucrose, and total soluble sugar was significantly higher in the leaves of ammonium-fed plants than nitrate-fed plants. The results indicate that nitrate/ammonium may serve as efficient regulators of oxalate accumulation owing to regulation of metabolism in rice leaves rather than oxalate downward transfer and root excretion, and that photosynthetic metabolism is not directly correlated with the regulation of oxalate accumulation in rice plants.     

Ammonium enhances the uptake, bioaccumulation, and tolerance of phenanthrene in cucumber seedlings

Aims

Phenanthrene is one of the ubiquitous, persistent organic pollutants commonly found in soil and sediments. The study will provide insight regarding the feasibility of nitrogen-assisted phytoremediation.

Methods

To study the effects of various nitrogen forms on cucumber seedling phenanthrene tolerance, hydroponic experiments were conducted in a greenhouse.

Results

Under phenanthrene stress, decreases in plant growth and biomass were more pronounced with a nitrate supply than with ammonium. In addition, phenanthrene concentrations in plants fed with ammonium were higher than those fed with nitrate. The reduction in plant protein and sugar, increases in nitrogen and phosphate concentrations, and increased activity of antioxidative enzymes may contribute to the phenanthrene stress response and adaptation. Higher peroxidase, superoxide dismutase, and catalase activities were found in ammonium-fed plants as compared to nitrate-fed plants under phenanthrene stress. Moreover, the reduction in soluble protein content and increases in phenanthrene transport and accumulation in non-photosynthetic organs may enable ammonium-fed plants to adapt more effectively to adverse conditions.

Conclusions

Overall, these results suggest that ammonium nutrition could provide a useful tool to improve the growth and adaption of plants under phenanthrene stress.     

Effects of low and high levels of magnesium on the response of sunflower plants grown with ammonium and nitrate

The effect of the nitrogen source (ammonium and nitrate) and its interaction with magnesium on various physiological processes was studied in sunflower plants (Helianthus annuusL.). Plants were grown in hydroponic culture with nitrate (5 mM) or ammonium (5 mM) and four concentrations of magnesium (0.1, 0.8, 5 and 10 mM). After 2 weeks, growth, gas exchange and fluorescence parameters, soluble carbohydrates, free amino acids, soluble protein and mineral elements were determined. Ammonium nutrition resulted in a reduction of dry matter accumulation, as well as in a decrease in the CO₂ assimilation. Moreover, ammonium-fed plants showed a greater content of free amino acids, soluble protein, Rubisco and anions, and a lower cation content, mostly Mg²⁺. The presence of high levels of Mg²⁺ in the nutrient solution containing NH₄ ⁺ resulted in a stimulation of growth and CO₂ assimilation to the levels observed in nitrate-fed plants. The lower photosynthetic rate of ammonium-fed plants grown with low level of magnesium does not seem to be due to a lower photosynthetic pigment content, or a deficiency in Photosystem II activity, or to lower Rubisco content. Hence, Rubisco activity or other enzymes involved in CO₂ fixation could have been affected in ammonium-fed plants. This revised version was published online in June 2006 with corrections to the Cover Date.     

Influence of NO3 - and NH4 + nutrition on fermentation,nitrate reductase activity and adenylate energy charge of roots of Carex pseudocyperus L. and Carex sylvatica Huds. exposed to anaerobic nutrient solutions

The adenylate energy charge, production of ethanol and lactate, and nitrate reductase activity were determined in order to study the influence of different nitrogen sources on the metabolic responses of roots of Carex pseudocyperus L. and Carex sylvatica HUDS. exposed to anaerobic nutrient solutions. Determination of adenylates was carried out by means of a modified HPLC technique. Total quantity of adenylates was higher in Carex pseudocyperus than in Carex sylvatica under all conditions. In contrast, the adenylate energy charge was only slightly different between the species and decreased more or less in relation to the applied nitrogen source under oxygen deficiency. The adenylate energy charge in roots of plants under nitrate nutrition showed a smaller decrease under anaerobic environmental conditions than plants grown with ammonium or nitrate/ammonium. Roots of nitrate-fed plants showed a lower ethanol and lactate production than ammonium/nitrate- and ammonium-fed plants. Ethanol production was higher in C. pseudocyperus, formation of lactate was lower compared to that in Carex sylvatica. The activity of enzymes involved in fermentation processes (ADH, LDH and PDC) was enhanced significantly after 24 hours of exposure to anaerobic nutrient solutions in roots of both species. The induction of these enzymes was only slightly influenced by different nitrogen supply. In vivo nitrate reductase activity increased almost 3-fold compared to the aerobic treatment in both species and overcompensated loss of NADH reoxidation capacity caused by decrease of ethanol and lactate development. Induction of in vitro nitrate reductase activity was enhanced 313% in C. pseudocyperus and 349% in C. sylvatica under anaerobic environmental conditions and nitrate supply. These results indicate that nitrate may serve as an alternative electron acceptor in anaerobic plant root metabolism and that the nitrate-supported energy charge may be due to an accelerated glycolytic flux resulting from a more effective NADH reoxidation capacity by nitrate reduction plus fermentation than by fermentation alone.Abbreviations ADH alcohol dehydrogenase - AEC adenylate energy charge - DMSO dimethyl sulfoxide - EDTA ethylen diamine tetraacetic acid - HPLC high performance liquid chromatography - LDH lactate dehydrogenase - NRA nitrate reductase activity - PCA perchloric acid - PDC pyruvate decarboxylase - PVP polyvinylpyrrolidone - PVPP polyvinylpolypyrrolidone - TCA trichloroacetic acid, Tris-tris(hydroxymethyl)aminomethane     

Chlorate Toxicity and Nitrate Reductase Activity in Tomato Plants

Chlorate damage was studied in tomato plants ( Lycopersicum esculentum cv. Moneymaker) that were supplied with a nitrogen-free nutrient solution or with a nutrient solution, containing either nitrate or ammonium as a nitrogen source. Damage was low in ammonium-fed plants and high in nitrate-fed plants and in nitrogen-less plants. Nitrate reductase activity could be detected in all treatments, although the activity was highest in the nitrate-fed plants.
The hypothesis that chlorate can be used as a substrate by the enzyme nitrate reductase in higher plants, was studied and proved to be true for the tomato plants, as was found earlier for Escherichia and Chlorella . The affinity of the enzyme for chlorate was lower than for nitrate, the K _m being 4 m M and 0.15 m M respectively. Induction of the enzyme by chlorate could not be detected. The enzyme activity was lowered in leaf discs after a 7 h treatment with chlorate and the inhibition was proportional to the chlorate concentration of the medium.
The results were discussed in terms of competition between nitrate and chlorate at the uptake and the enzyme site and with regard to a possible influence of chlorate on synthesis and breakdown of the enzyme.     

Changes in the Morphology of Roots and Leaves of Carob Seedlings Induced by Nitrogen Source and Atmospheric Carbon Dioxide

Carob seedlings were grown hydroponically for 9 weeks under360 and 800 µl l^-1CO₂. One of two nitrogen sources, nitrateor ammonium, was added to the nutrient medium at concentrationsof 3 mol m^-3. Root systems of the developing plants suppliedwith nitrate compared to those supplied with ammonium were characterizedby:(a)more biomass on the lower part of the root;(b)fewer lateralroots of first and second order;(c)longer roots;(d)higher specificroot length;(e)a smaller root diameter. The morphology of theroot systems of nitrate-fed plants changed in the presence ofelevated carbon dioxide concentrations, resembling, more closely,that of ammonium-fed plants. Total leaf area was higher in ammonium-than in nitrate-fed plants. Nitrate-fed plants had greater totalleaf area in the presence of high carbon dioxide than in normalCO₂, due to an increase in epidermal cell size that led to developmentof larger leaflets with lower stomatal frequency. The observedchanges in the morphology of roots and shoots agreed with theresults observed for total biomass production. Nitrate-fed plantsincreased their biomass production by 100% in the presence ofelevated CO₂compared to 15% in ammonium-fed plants, indicatingthat the response of carob to high CO₂concentrations is verydependent on the nitrogen source. Under elevated CO₂, nitrate-grownplants had a larger content of sucrose in both roots and shoots,while no significant difference was observed in the contentof sucrose in ammonium-grown plants, whether in ambient or enrichedcarbon dioxide. Hence, the differences in soluble carbohydratecontents can, at least partly, account for differences in rootand shoot morphology.Copyright 1997 Annals of Botany Company Ceratonia siliquaL.; carob; ammonium; carbohydrate; carbon dioxide; nitrate; morphology; sucrose     

Effect of salinity and different nitrogen sources on the activity of antioxidant enzymes and indole alkaloid content in Catharanthus roseus seedlings

The activities of antioxidant enzymes viz. glutathione reductase, GR; superoxide dismutase, SOD; peroxidase, POD; catalase, CAT and glutathione-S-transferase, GST and alkaloid accumulation were investigated in leaf pairs (apical, middle, basal) and in roots of Catharanthus roseus seedlings under the conditions of different nitrogen sources (20 mM KNO(3) and 2 mM NH(4)Cl) and salinity, in the absence (non-saline control) and in the presence of 100 mM NaCl in the nutrient solution. Salinity caused a reduction in plant biomass. The biomass production of ammonium-fed plants was lower than that of nitrate-fed plants. The antioxidant enzymes exhibited higher activity in saline-treated plants. Changes in antioxidant enzyme activity caused by different nitrogen sources differed in all leaf pairs, as well as in roots of C. roseus. Ammonium-fed plants showed higher CAT, GR and GST activity in leaf pairs as well as in roots, while POD and SOD activity were higher in nitrate-fed plants. Higher peroxidase activity concomitant with the increased accumulation of alkaloid was found in all leaf pairs, as well as in roots of C. roseus of NO(3)(-) fed plants as compared to NH(4)(+) fed plants.     

Interaction of nitrogen nutrition and salinity in Grey poplar (Populus tremula x alba)

Salinity represents an increasing environmental problem in managed ecosystems. Populus spp. is widely used for wood production by short-rotation forestry in fertilized plantations and can be grown on saline soil. Because N fertilization plays an important role in salt tolerance, we analysed Grey poplar (Populus tremula x alba, syn. Populus canescens) grown with either 1 mM nitrate or ammonium subjected to moderate 75 mM NaCl. The impact of N nutrition on amelioration of salt tolerance was analysed on different levels of N metabolism such as N uptake, assimilation and N (total N, proteins and amino compounds) accumulation. Na concentration increased in all tissues over time of salt exposure. The N nutrition-dependent effects of salt exposure were more intensive in roots than in leaves. Application of salt reduced root increment as well as stem height increase and, at the same time, increased the concentration of total amino compounds more intensively in roots of ammonium-fed plants. In leaves, salt treatment increased concentrations of total N more intensively in nitrate-fed plants and concentrations of amino compounds independently of N nutrition. The major changes in N metabolism of Grey poplar exposed to moderate salt concentrations were detected in the significant increase of amino acid concentrations. The present results indicate that N metabolism of Grey poplar exposed to salt performed better when the plants were fed with nitrate instead of ammonium as sole N source. Therefore, nitrate fertilization of poplar plantations grown on saline soil should be preferred.     

Alternative pathway respiration is associated with ammonium ion sensitivity in spinach and pea plants

Spinach and pea plants were grown in hydroponic culture with nitrate orammonium salts as the nitrogen source. Dry matter accumulation andphotosynthetic rate declined in spinach plants fed with ammonium salts, whereasthey did not change in pea plants compared with nitrate-fed plants. Measurementof organic nitrogen and free amino acid content showed that ammonium ions wereassimilated in shoots in spinach plants and in roots in pea plants. Ammoniumionnutrition led to a decline in starch content in both species. Organic acidsincreased in roots of pea plants fed with ammonium ions whereas they declinedinspinach plants. In both species ammonium ions increased root respiration ratebut the contribution of both routes (cytochromic and alternative pathway) tothis increase was different depending on the species. In spinach plants,ammonium ions increased the cytochromic path and decreased the alternativepathway, whereas in pea plants both routes were stimulated mainly through thealternative pathway. The differences in the sensitivity to ammonium ionsbetweenboth species are discussed in terms of differences in the availability of Cskeletons and energy, which could be due in part to differences in the capacityto stimulate the alternative pathway.     

Ammonium Nutrition as a Strategy for Cadmium Mobilisation in the Rhizosphere of Sunflower

Ammonium nutrition of higher plants results in rhizosphere acidification due to proton excretion by root cells. The acidification induced by ammonium-fed plants can be exploited to promote a localised metal mobilisation in neutral to alkaline polluted soils and therefore to improve phytoextraction. The effects of ammonium uptake by sunflower (Helianthus annuus L.) plants on the external medium pH, aerial and root growth and tolerance to soluble Cd were studied in hydroponic culture. The ammonium-fed sunflowers induced a strong acidification of the solution and, compared to the nitrate-fed sunflowers, a small modification in mineral nutrition and a different Cd partitioning between root and shoot. Moreover, ammonium nutrition was found to induce a great mobilisation of a sparingly soluble form of cadmium (CdCO₃). A pot experiment studied the ability of different ammonium-based fertilisers (ammonium sulphate, ammonium thiosulphate, urea) to modify bulk and rhizo-soil pH, compared to the effect of calcium nitrate and to the unfertilised soil. Furthermore, in order to promote the persistence of ammonium in soil, a combined treatment of ammonium sulphate and DMPP, a nitrification inhibitor, was tested. Soil pH was strongly modified by chemical and biological processes involved in fertiliser transformations. In particular, due to nitrification, all ammonium-based treatments showed a bulk soil acidification of over 1.5 pH units and a relative increase in rhizo-soil pH as a consequence of nitrate uptake. The treatment with DMPP showed an opposite trend with a lower pH in rhizo-soil than in bulk soil. The ability of ammonium-fed plants to mobilise heavy metals from the non-labile pool was studied in another pot experiment using three soils with different properties and at different degree and type of heavy metal contamination. Whatever the soil, the metal concentrations in shoots were higher in plants fed with ammonium (ammonium sulphate plus DMPP treatment). Our results support the hypothesis that ammonium nutrition with nitrification inhibitors is a viable strategy to improve heavy metals phytoextraction while protecting bulk soil from acidification and presumably from metal leaching. An erratum to this article is available at .     

Inhibition of endogenous urease activity by NBPT application reveals differential N metabolism responses to ammonium or nitrate nutrition in pea plants: a physiological study

Background and aims

Urea is the predominant form of N applied as fertilizer to crops, but it is also a significant N metabolite of plants themselves. As such, an understanding of urea metabolism in plants may contribute significantly to subsequent N fertilizer management. It currently appears that arginase is the only plant enzyme that can generate urea in vivo. The aim of this work was, therefore, to gain a more in-depth understanding of the significance of the inhibition of endogenous urease activity and its role in N metabolism depending on the N source supplied.

Methods

Pea (Pisum sativum cv. Snap-pea) plants were grown with either ammonium or nitrate as the sole N source in the presence or absence of the urease inhibitor NBPT.

Results

When supplied, NBPT is absorbed by plants and translocated from the roots to the leaves, where it reduces endogenous urease activity. Different N metabolic responses in terms of N-assimilatory enzymes and N-containing compounds indicate a different degree of arginine catabolism activation in ammonium- and nitrate-fed plants.

Conclusions

The arginine catabolism is more highly activated in ammonium-fed plants than in nitrate-fed plants, probably due to the higher turnover of substrates by enzymes playing a key role in N recycling and remobilization during catabolism and in early flowering and senescence processes, usually observed under ammonium nutrition.     

Ammonium and nitrate nutrition inPlantago lanceolata L. andPlantago major L. ssp.major

With the aims (1) to test whether the different natural occurrence of twoPlantago species in grasslands is explained by a different preference of the species for nitrate or ammonium; (2) to test whether the different occurrence is explained by differences in the flexibility of the species towards changes in the nitrogen form; (3) to find suitable parameters as a tool to study ammonium and nitrate utilization of these species at the natural sites in grasslands, plants ofPlantago lanceolata andP. major ssp.major were grown with an abundant supply of nitrate, ammonium or nitrate+ammonium as the nitrogen source (0.5 mM). The combination of ammonium and nitrate gave a slightly higher final plant weight than nitrate or ammonium alone. Ammonium lowered the shoot to root ratio inP. major. Uptake of nitrate per g root was faster than that of ammonium, but from the mixed source ammonium and nitrate were taken up at the same rate. In vivo nitrate reductase activity (NRA) was present in both shoot and roots of plants receiving nitrate. When ammonium was applied in addition to nitrate, NRA of the shoot was not affected, but in the root the activity decreased. Thus, a larger proportion of total NRA was present in the shoot than with nitrate alone. In vitro glutamate dehydrogenase activity (GDHA) was enhanced by ammonium, both in the shoot and in the roots.In vitro glutamine synthetase activity (GSA) was highest in roots of plants receiving ammonium. Both GDHA and GSA were higher inP. lanceolata than inP. major. The concentration of ammonium in the roots increased with ammonium, but it did not accumulate in the shoot. The concentration of amino acids in the roots was also enhanced by ammonium. Protein concentration was not affected by the form of nitrogen. Nitrate accumulated in both the shoot and the roots of nitrate grown plants. When nitrate in the solution was replaced by ammonium, the nitrate concentration in the roots decreased rapidly. It also decreased in the shoot, but slowly. It is concluded that the nitrogen metabolism of the twoPlantago species shows a similar response to a change in the form of the nitrogen source, and that differences in natural occurrence of these species are not related to a differential adaptation of nitrogen metabolism towards the nitrogen form. Suitable parameters for establishing the nitrogen source in the field are thein vivo NRA, nitrate concentrations in tissues and xylem exudate, and the fraction of total reduced nitrogen in the roots that is in the soluble form, and to some extent thein vitro GDHA and GSA of the roots. Grassland Species Research Group. Publ. no 118.     

Interaction of sulfur and nitrogen nutrition in tobacco (Nicotiana tabacum) plants: significance of nitrogen source and root nitrate reductase

Abstract: The significance of root nitrate reductase for sulfur assimilation was studied in tobacco (Nicotiana tabacum) plants. For this purpose, uptake, assimilation, and long-distance transport of sulfur were compared between wild-type tobacco and transformants lacking root nitrate reductase, cultivated either with nitrate or with ammonium nitrate. A recently developed empirical model of plant internal nitrogen cycling was adapted to sulfur and applied to characterise whole plant sulfur relations in wild-type tobacco and the transformant. Both transformation and nitrogen nutrition strongly affected sulfur pools and sulfur fluxes. Transformation decreased the rate of sulfate uptake in nitrate-grown plants and root sulfate and total sulfur contents in root biomass, irrespective of N nutrition. Nevertheless, glutathione levels were enhanced in the roots of transformed plants. This may be a consequence of enhanced APR activity in the leaves that also resulted in enhanced organic sulfur content in the leaves of the tranformants. The lack of nitrate reductase in the roots in the transformants caused regulatory changes in sulfur metabolism that resembled those observed under nitrogen deficiency. Nitrate nutrition reduced total sulfur content and all the major fractions analysed in the leaves, but not in the roots, compared to ammonium nitrate supply. The enhanced organic sulfur and glutathione levels in ammonium nitrate-fed plants corresponded well to elevated APR activity. But foliar sulfate contents also increased due to decreased re-allocation of sulfate into the phloem of ammonium nitrate-fed plants. Further studies will elucidate whether this decrease is achieved by downregulation of a specific sulfate transporter in vascular tissues.     

Osmotically Induced Proline Accumulation in <Emphasis Type="Italic">Lotus Corniculatus</Emphasis> Leaves is Affected by Light and Nitrogen Source

Proline accumulation in osmotically stressed leaves of Lotus corniculatus was stimulated by increasing light intensity (photon fluence density, PFD). Treatment with propanil limited proline accumulation in response to light and osmotic stress, indicating a dependence of proline synthesis on photosynthetic NADPH. Drought stress induced proline accumulation in L. corniculatus both in nitrate-fed plant (NFP) and ammonium-fed plants (AFP), although higher proline concentration was observed in AFP than in NFP after 24 h of drought stress. Changes in proline accumulation induced by drought stress in plants grown under different nitrogen regimes could not be explained by changes of either total protein or amino acids, consistent with specifically altered regulation of proline synthesis. Under control conditions, alanine, aspartate and glutamate were the predominant amino acids in NFP; conversely, in AFP, arginine and ornithine were the predominant amino acids. Only the NFP regime showed changes in the concentrations of specific amino acids under drought stress a decrease in alanine, aspartate and glutamate and increased gama-aminobutyric acid. In AFP and especially NFP, proline accumulation under osmotic stress was associated with increased ornithine amino transferase activity. An increase of both activity and protein of ferredoxin-dependent glutamate synthase was observed in osmotic-stressed NFP; inversely both decreased in drought-stressed AFP. PFD and nitrogen source are therefore shown to be regulators of proline accumulation in L. corniculatus osmotically stressed plants.     

Nitrogen assimilation and transport in carob plants

Most of the nitrate reductase activity (80%;) in carob ( Ceratonia siliqua L. cv. Mulata) is localised in the roots. The nitrate concentration in the leaves is relatively low compared to that in the roots, suggesting that nitrate influx into the leaf may be a major factor limiting the levels of nitrate reductase in the shoot. Transport of nitrate from root to shoot appears limited by the entrance of nitrate into the xylem. In order to study this problem, we determined the nitrate concentrations and nitrate reductase activities along the roots of nitrate-grown plants, as well as the composition of the xylem sap and the nitrate levels in the leaves. Some of the the bypocotyl, in order to bypass the loading of nitrate into the xylem of the roots. The results show that the loading of nitrate into the xylem is a limiting step.
The cation and anion concentrations of nitrate- and ammonium-fed plants were similar, showing almost no production of organic anions. In both nitrate- and ammonium-fed plants, the transport of nitrogen from root to shoot was in the form of organic nitrogen compounds. The nitrate reductase activity in the roots was more than sufficient to explain all the efflux of OH⁻ into the root medium of nitrate-fed plants. In carob plants the K-shuttle may thus be operative to a limited extent only, corresponding to between 11 and 27%; of the nitrate taken up. Potassium seems to be the cation accompanying stored nitrate in the roots of carob seedlings, since they accumulate nearly stoichiometric amounts of K⁺ and NO⁻₃.