Changes in the Activity of Antioxidant Enzymes in Wheat Leaves and Roots as a Function of Nitrogen Source and Supply

The activity of enzymes participating in the systems of antioxidant protection was assayed in the second leaf and roots of 21-day-old wheat seedlings (Triticum aestivum L.) grown in a medium with nitrate (NO^– ₃ treatment), ammonium (NH⁺ ₄ treatment), or without nitrogen added (N-deficiency treatment). The activities of superoxide dismutase (SOD), peroxidase, ascorbate peroxidase, glutathione reductase, and catalase in the leaves and roots of the NH⁺ ₄ plants was significantly higher than in the plants grown in the nitrate medium. The activity of SOD decreased and ascorbate peroxidase markedly increased in leaves, whereas the activity of ascorbate peroxidase increased in the roots of N-deficient plants, as compared to the plants grown in nitrate and ammonium. Low-temperature incubation (5°, 12 h) differentially affected the antioxidant activity of the studied plants. Whereas leaf enzyme activities did not change in the NH⁺ ₄ plants, the activities of SOD, peroxidase, ascorbate peroxidase, and catalase markedly increased in the NO^– ₃ plants. In leaves of the N-deficient plant, the activity of SOD decreased; however, the activity of other enzymes increased. In response to temperature decrease, catalase activity increased in the roots of NO^– ₃ and NH⁺ ₄-plants, whereas in the N-deficient plants, the activity of peroxidase increased. Thus, in wheat, both nitrogen form and nitrogen deficiency changed the time-course of antioxidant enzyme activities in response to low temperature.     

Changes in amino acid content of an algal feed species (Navicula sp.) and their effect on growth and survival of juvenile abalone (Haliotis rubra)

The growth and survival of juvenile Haliotis rubra, when fed with the diatom Navicula sp. cultured in f/2 medium containing combined nitrogen at 24.71 mg NO₃-N L^–1 (high), 12.35 mg NO₃-N L^–1 (standard) or 2.47 mg NO₃-N L^–1 (low), were compared in a 33-day trial. The alga in the low nitrogen medium contained 37% less total amino acid than that in the high and standard nitrogen media. There was a slightly greater reduction in essential amino acids (40%) compared to non-essential amino acids (35%). Juvenile abalone feeding on Navicula grown in medium with low nitrate and lower total amino acid content grew more slowly than when fed on the same species grown in standard or higher nitrogen medium with a higher amino acid content. The growth rate of juveniles was highest (43 m d^–1) in the high nitrate treatment followed (40 m d^–1) by the standard nitrate treatment and lowest (31 m d^–1) in the low nitrate treatment. The survival of the juveniles was also effected by the diet. Survival was better in the high and standard nitrogen media (88%) than the low nitrogen medium (75%). The results suggest that in order to achieve uniformity in nutritional quality of diatoms and good growth of abalone juveniles in commercial abalone nurseries, the nitrogen concentration in tanks should be monitored and additional nitrate added to provide an optimum concentration of between 2 and 12 mg NO₃-N L^–1.     

Growth, water use efficiency, and proline content of hydroponically grown tomato plants as affected by nitrogen source and nutrient concentration

Tomato plants (Lycopersicon esculentum Mill. cv. Counter) were grown in 12-L polyethylene containers in aerated and CaCO₃-buffered nutrient solutions containing different concentrations of complete stock solutions with either nitrate (stock solution N) or ammonium (stock solution A) as the only nitrogen source (X1 = standard concentration with 5 mM NO₃ ^–-N or NH₄ ⁺-N, and X3, X5.5, X8 and X11 = 3, 5.5, 8, 11 times the standard concentration), or a mixture of both stock solutions (N:A ratio = 100:0, 75:25, 50:50, 25:75, 0:100) at moderate nutrient concentration (X3). Total dry matter production and fruit dry weight were only slightly affected by increasing nutrient concentration if nitrate was supplied as the sole nitrogen source. Compared to nitrate, ammonium nitrogen caused a decrease in total dry weight (32–86% between X1 and X11), but led to an increase in both total dry weight and fruit dry weight (11% and 30%) at low concentration if supplied in addition to nitrate nitrogen (N:A ratio = 75:25). Dry matter partitioning in plants was affected by the strength of the nutrient solution, but even more by ammonium nitrogen. Fruits accumulated relatively less dry matter than did the vegetative parts of tomato plants when supplied with nutrient solutions containing ammonium as the only nitrogen source (fruit dry weight to total dry weight ratio 0.37 and 0.15 at low and high nutrient concentration), while nitrate nitrogen rather supported an increase in dry matter accumulation in the reproductive organs (fruit dry weight to total dry weight ratio 0.39–0.46). The water use efficiency (WUE) was only slightly affected by the strength of the nutrient solutions containing nitrate nitrogen (2.9–3.4 g DW (kg H₂O)^–1), while ammonium nitrogen led to a decrease in WUE from 2.4 to 1.3 g DW (kg H₂O)^–1at low (X1) and high (X11) nutrient concentration, respectively. The proline content of leaves fluctuated (0.1–5.0 mol (g fresh weight)^–1) according to nutrient concentration and global radiation, and reflected enhanced sensitivity of plants to these potential stress factors if ammonium was the predominant N source supplied. It was concluded, that proline is a reliable indicator of the environmental stress imposed on hydroponically grown tomato plants.     

Accumulation of Oxalate in Tissues of Sugar-beet, and the Effect of Nitrogen Supply

In field-grown sugar-beet concentration of insoluble oxalatewas low in roots and high (about 12 per cent of ethanol insolublematerial) in leaves, and for a particular leaf the concentrationincreased continuously during its life. Of the insoluble oxalate,15–30 per cent was present as the magnesium salt and theremainder as the calcium salt. Oxalate contents of plants grownin culture solutions with nitrate as nitrogen source were similarto those of plants grown in soil, but when nitrogen was suppliedas ammonium sulphate or ammonium nitrate both soluble and insolubleoxalate were low. Plants grown in soil with regular additionsof ammonium sulphate or ammonium nitrate also had very low concentrationsof soluble oxalate although insoluble oxalate was only slightlylower than with nitrate nitrogen. Disks of root or leaf tissuewashed for several days in distilled water lost insoluble oxalatebut when washed in tap water insoluble oxalate increased morethan twofold. Addition of calcium and nitrate to the distilledwater caused an increase of insoluble oxalate, while additionof potassium caused a decrease. Use of ¹⁴C labelled oxalateand washing experiments showed that oxalate can be metabolizedby tissue disks and so is not necessarily a final product ofmetabolism. The accumulation of oxalate appears to be connectedwith the assimilation of nitrate and the preservation of thecation-anion balance of the plant.     

Heterogeneity of Maize Root Mitochondria from Plants Grown in the Presence of Ammonium

Mitochondria isolated from root tissue of maize plants grown on a modified Knop solution containing 10.9 mM nitrate ± 7.2 mM ammonium were purified on the discontinuous Percoll density gradient with polyvinylpyrrolidone (PVP) added. The presence of PVP allowed separation of several mitochondrial fractions of a different density. Contrary to mitochondria isolated from plants grown in the presence of nitrate alone, revealing only two fractions, the mitochondria from NH₄ ⁺/NO₃ ^–-plants were distributed in four fractions. Total amount of mitochondria, as well as specific activities of some nitrogen metabolism enzymes and tricarboxylic acid (TCA) cycle enzymes of all mitochondrial fractions, and respiratory activities of two lower density fractions isolated from plants grown on mixed nitrogen were higher in comparison to mitochondria from nitrate-grown plants.     

Nitrate Assimilation during Vegetative Regrowth of Alfalfa

Dry matter accumulation, nitrate reductase activity of various organs, nitrate accumulation, nitrogen derived from nitrate, and nitrogen content were studied during 17 days of vegetative regrowth of harvested (detopped) alfalfa (Medicago sativa L.). Seedlings were grown in the glasshouse and treated with 0, 40, and 80 kilograms N per hectare applied as K¹⁵NO₃ to determine whether reduced nitrogenase activity after shoot harvest limited vegetative regrowth. The role of nodules in reducing NO₃⁻ during this period of low nitrogenase activity was also investigated.     

Response of Eggplant to Nitrogen Supply: Molybdenum-Nitrate Relationships

Greenhouse experiments were conducted in two years (1993–1994) with eggplants supplied with 1, 2 or 4 mM NH₄NO₃ as the N source in order to determine its influence on molybdenum (Mo) and nitrate (NO₃ ^–) content in leaf blades, petioles, and fruits as well as leaf nitrate reductase (NR) activity. The results reveal that 2 and 4 mM NH₄NO₃ altered shoot Mo distribution and thus affecting the NR activity.     

Persistence and competitiveness of threeBradyrhizobium japonicum inoculant strains in clay loam Nile valley soil

The nitrogen contribution from the shoot and root system of symbiotically grown leucaena was evaluated in a field experiment on an Alfisol at IITA in Southern Nigeria. Maize in plots that received prunings from inoculated leucaena contained more N and grain yield was increased by 1.9 t.ha.^–1. Large quantities of nitrogen were harvested with leucaena prunings (300 kg N ha^–1 in six months) but the efficiency of utilization of this nitrogen by maize was low compared to inorganic N fertilizer (ammonium sulphate) at 80 kg N ha^–1. Maize yield data indicated that nitrogen in leucaena prunigs was 34 and 45% as efficient as 80 kg N ha^–1 of (NH₄)₂SO₄ for uninoculated and inoculated plants with Rhizobium IRc 1045, respectively. In plots where the prunings were removed, the leaf litter and decaying roots and nodules contributed N equivalent of 32 kg ha^–1. Twenty-five kg ha^–1 was the inorganic N equivalent from nitrogen fixed symbiotically by leucaena when inoculated with Rhizobium strain IRc 1045. Application of prunings from inoculated leucaena resulted in higher soil ogranic C, total N, pH and available NO₃.     

The effect of Nitrogen on the size of spring barley root system as determined by means of its electric capacity

The size of the spring barley root system was studied on the basis of its electric capacity in plants grown in nutrient solutions either lacking or containing nitrogen in the form of nitrate or ammonium. Root electric capacity changed in dependence on nutrition from Day 12 after emergence, when F values increased in the root systems of plants exposed to nitrate and ammonium salts. In plants grown in H₂O, the values of electric capacity statistically significantly decreased on Days 15 to 17, in plants grown in PK solution lacking nitrogen on Day 20. Root electric capacity of plants grown in full nutrient solution gradually increased on Day 18 after emergence. Then a marked increase in root electric capacity values followed with no statistically significant differences between NH₄ ⁺ and NO₃ ^- nutrition. Nitrate nutrition of barley plants only resulted in an increased root to shoot mass ratio.     

Effect of two nutrient solution temperatures on nitrate uptake,nitrate reductase activity,NH4+ concentration and chlorophyll a fluorescence in rose plants

The effect of two nutrient solution temperatures (cold (10 °C) and warm (22 °C)) during two flowering events of rose plants (Rosa × hybrida cv. Grand Gala) were examined by measuring chlorophyll (Chl) a fluorescence, ammonium (NH₄⁺) content and nitrate reductase (NR) activity in four different leaf types, that is, external and internal leaves of bent shoots and lower and upper leaves of flowering stems. Besides, nitrate (NO₃^?) uptake and water absorption, total nitrogen (N) concentration in the plant, dry biomass, and the ratios of shoot/root and thin-white roots/suberized-brown roots were determined. Generally, cold solution increased NO₃^? uptake and thin-white roots production but decreased water uptake, so plants grown at cold solution had to improve their NO₃^? uptake mechanisms to obtain a higher amount of nutrient with less water absorption than plants grown at warm solution. The higher NO₃^? uptake can be related to an increase in NR activity, NH₄⁺ content and total N concentration at cold solution. Nutrient solution temperature also had an effect on the photosynthetic apparatus. In general terms, the effective quantum yield (?_PSII) and the fraction of open PSII reaction centres (q_L) were higher in rose plants grown at cold solution. These effects can be associated to a higher NO₃^? uptake and total N concentration in the plants and were modulated by irradiance throughout all the experiment. Plants could adapt to cold solution by enhancing their metabolism without a decrease in total dry biomass. Nevertheless, the effect of nutrient solution temperature is not simple and also affected by climatic factors.     

Aspects of nitrogen metabolism in the rice seedling

The effects of nitrogen source NO₃⁻ or NH₄⁺ on nitrogen metabolism during the first 2 weeks of germination of the rice seedling (Oryza sativa L., var. IR22) grown in nutrient solution containing 40 μg/ml N were studied. Total, soluble protein, and free amino N levels were higher in the NH₄⁺-grown seedling, particularly during the 1st week of germination. Asparagine accounted for most of the difference in free amino acid level, in both the root and the shoot. Nitrate and nitrite reductase activities were present mainly in the shoot and were higher in the NO₃⁻-grown seedling, whereas the activity of glutamate dehydrogenase and glutamine synthetase in the root tended to be lower than that of the NH₄⁺-grown seedling during the 1st week of germination. Glycolate oxidase and catalase activities were present mainly in the shoot. Maximum activity of the above five enzymes occurred 7 to 10 days after germination. Differences in the zymograms of nitrate reductase, glutamate dehydrogenase, and catalase were mainly between shoot and root and not from N source. Nitrite reductase bands were observed only in plants grown in plants grown in NO₃⁻.     

The role of nitrate in osmoregulation of Italian ryegrass

Summary The role of nitrate in osmotic control was studied with Italian ryegrass grown in a nutrient solution in a climate room. Quantum-flux density, osmotic potential of the nutrient solution and availability of nitrate and chloride were varied independently. Plants at high quantum flux density (650 mol m^–2 s^–1) had a lower osmotic potential, a higher carbohydrate concentration and a lower nitrate concentration than plants at low quantum flux density (310 mol m^–2 s^–1), the decrease in nitrate concentration was osmotically equivalent to the increase in carbohydrate concentration. When nitrate in the nutrient solution was partly replaced by chloride, the chloride taken up substituted an equivalent part of the nitrate in the plant. It is concluded that nitrate plays a role in osmoregulation of the plant and compensates for a shortage of other solutes.     

The possibility of predicting solute uptake and plant growth response from independently measured soil and plant characteristics

Summary The growth and nitrogen uptake response of rape plants to nitrate concentration at the root surface were studied in solution culture in a controlled environment cabinet over a period of 24 days. NO₃ ^– was supplied at the rates of 10^–5 M, 5×10^–5 M, 10^–4 M, 10^–3 M and 10^–2 M in solution and was maintained near these levels.With increasing mean N concentration in the tissues, the relative growth rate and leaf area ratio increased and unit leaf rate decreased slightly. Values of all three growth parameters decreased with plant age.The shoot: root dry weight ratios and their N content ratios increased with increasing mean per cent N in the plant. The length or surface area per unit dry weight of roots was correlated negatively with per cent N and positively with age.The maximum mean inflow of nitrate to rape roots decreased sharply with age. The concentration at which half maximal mean inflow was attained was 3.44×10^–5 M NO₃ ^–.     

The use of different soil nitrogen sources by young Norway spruce plants

 Three-year-old Norway spruce trees were planted into a low-nitrogen mineral forest soil and supplied either with two different levels of mineral nitrogen (NH₄NO₃) or with a slow-release form of organic nitrogen (keratin). Supply of mineral nitrogen increased the concentrations of ammonium and nitrate in the soil solution and in CaCl₂-extracts of the rhizosphere and bulk soil. In the soil solution, in all treatments nitrate concentrations were higher than ammonium concentrations, while in the soil extracts ammonium concentrations were often higher than nitrate concentrations. After 7 months of growth, ¹⁵N labelled ammonium or nitrate was added to the soil. Plants were harvested 2 weeks later. Keratin supply to the soil did not affect growth and nitrogen accumulation of the trees. In contrast, supply of mineral nitrogen increased shoot growth and increased the ratio of above-ground to below-ground growth. The proportion of needle biomass to total above-ground biomass was not increased by mineral N supply. The atom-% ¹⁵N was higher in younger needles than in older needles, and in younger needles higher in plants supplied with ¹⁵N-nitrate than in plants supplied with ¹⁵N-ammonium. The present data show that young Norway spruce plants take up nitrate even under conditions of high plant internal N levels. Received: 1 April 1998 / Accepted: 9 October 1998     

Nitrogen and base cation uptake in seedlings of Acer pseudoplatanus and Calamagrostis villosa exposed to an acidified environment

The effects of solution acidity and form of nitrogen on net nutrient uptake rates in Acer pseudoplatanus and Calamagrostis villosa seedlings were examined as part of a complex ecological study. Uptake rates were measured by the depletion method under controlled conditions (temperature 20 °C, irradiance 400 mol m^–2 s^–1 PAR) from a nutrient solution containing 1.5 mM nitrogen in the form of nitrate or ammonium or an equimolar mixture of both. The solution acidity was kept constant at pH 5.5 (control treatment), 4.5 or 3.5 (low pH treatments). Strongly acid pH decreased or stopped the uptake rates of NO₃ ^–, Mg²⁺ and Ca²⁺, but the uptake of NH₄ ⁺ was not changed in both species. Ammonium ions reduced the uptake rate of NO₃ ^– in Acer but increased the uptake rate in Calamagrostis. Ammonium as the sole source of nitrogen had a strong negative impact on the uptake rates of calcium and magnesium and this effect was independent of the media acidification usually connected with NH₄ ⁺ uptake and assimilation. However, the negative effect of ammonium ions on the base cation uptake was more pronounced at low pH values.     

Phosphate availability in combination with nitrate availability affects root yield and chicon yield and quality of Belgian endive (Cichorium intybus)

This study investigated the effects of nitrate and phosphate nutrition on chicory tap root development and chicon quality. Plants of chicory (Cichorium intybus flash) were grown on four concentrations of nitrate and phosphate: 3 mM NO₃ / 1 mM PO ₄ ^3– , high N and high P (control plants, N / P); 3 mM NO ₃ ^– / 0.05 mM H₂PO^3– ₄, high N and low P (N / p); 0.6 mM NO₃ / 1 mM PO ₄ ^3– , low N and high P (n / P); 0.6 mM NO ₃ ^– / 0.05 mM PO ₄ ^3– , low N and low P (n / p). The results suggested that, nitrogen limitation had the greatest impact on the shoot/root dry weight ratio. Only small changes in the shoot/root dry weight could be attributed to P limitation alone. Compared with the control, N limitation caused a marked increase in root SST activity (sucrose sucrose fructosyl transferase, the enzyme responsible for fructan synthesis in roots), the effect of P limitation on SST activity was less pronounced. The activity of SS (sucrose synthase) was also noticeably elevated at the early sample data by N limitation. N and P uptake were estimated by the amount of N and P accumulated by the whole plant during the vegetative period. With N limitation, P accumulation was decreased by 40-60% over the experimental period. The effects of P limitation on N accumulation were more variable, N uptake was 60% lower than the control during the tuberizing period (107 days after sowing). With N limitation, P concentrations in roots were lowered by 20-25%. With P limitation, total N concentration in roots decreased by 50% relative to the control, while nitrate concentration was increased more than 8 fold. These effects were detected only at 107 DAS. The amino acid content of roots was not affected by P limitation, however, N limitation altered strongly total amino acids. P limitation did alter the relative amino acid composition of roots early in the vegetative period: Roots harvested at the end of vegetative period were forced in the dark to produce an etiolated bud, the edible chicon. High N and high P fertility (N/P) were associated to a poor chicon yield and quality. However the presence of low P during vegetative growth moderates adverse effects of high nitrate and greatly improved chicon yeild and quality.     

High frequency plant regeneration from thin cell layer explants of Brassica napus

Explants composed of the epidermis and 4–9 layers of subepidermal cells were excised from internodes of Brassica napus L. ssp. oleifera cv. Westar and cultured on modified Murashige and Skoog (MS) medium. The three or four terminal internodes excised from plants at an early stage (before any flower buds had opened) were shown to be the best explant source. Both cytokinin and auxin were required for induction of shoot organogenesis. Of six auxins tested, only naphthaleneacetic acid (NAA) was effective in shoot bud initiation. All four cytokinins tested (when associated with 0.5 mgl^-1 NAA) promoted organogenesis, but at differing frequencies. The highest shoot induction frequency was obtained at 10–15 mgl^-1 benzyladenine (BA). The organogenic response was strongly affected by the nitrogen content of the medium. The best response was observed when NO₃ ^- was the sole nitrogen source (supplied as KNO₃) in the range 30–90 mM. Sucrose and glucose were equally supportive in shoot regeneration with the optimal levels at 0.12 M and 0.15 M, respectively. Shoots were rooted on medium free of growth regulators and mature plants were grown in the greenhouse. Plants were also recovered from leafy structures which differed morphologically and histologically from shoot buds.     

Effects of pH and nitrogen on cadmium uptake in potato

This study investigated the effects of pH and nitrogen form and concentration on cadmium (Cd) uptake by potato (Solanum tuberosum L.) grown in hydroponic culture. Potato plants grown in a pH-buffered nutrient solution for 10 d were exposed for 24 h to 25 nM CdCl₂ labelled with ¹⁰⁹Cd. Plants showed a significantly higher Cd uptake and accumulation at pH 6.5 than at pH 4.5 and 5.5. Nitrogen supplied as nitrate (NO₃ ^?) generally resulted in a higher Cd uptake and accumulation than N supplied as ammonium (NH₄ ⁺). This effect was most pronounced at pH 6.5. The N concentration increasing from 6.5 to 26 mM resulted in a decreased Cd influx when either NO₃ ^? or NH₄ ⁺ was used. Cd translocation to the shoot was increased when NO₃ ^? was used as the sole N source. In conclusion, pH had a strong influence on Cd uptake by roots and N form is especially important for Cd translocation within the potato plant.     

Importance of myo-inositol,calcium, and ammonium for the viability and division of tomato (Lycopersicon esculentum) protoplasts

Plants from four cultivars of Lycopersicon esculentum were grown under different conditions, in controlled environment chambers. Low light intensity, long photoperiod (16 h), 25° C/17°C temperature alternance (day/night) were found to be the most convenient conditions for obtaining viable protoplasts. The use of myo-inositol as an osmoticum in the digestion medium and the adjustment of the pH to 6.5, instead of the usual 5.8, for this medium increased the yield of viable protoplasts and enhanced their stability. Under these conditions neither pretreatment (dark and cold treatments), nor preplasmolysis of leaf tissues, were required before protoplast isolation. The concentrations of ammonium nitrate, calcium chloride, myo-inositol, and sucrose were found to be critical for the success of protoplast culture. A medium containing 5 mM ammonium nitrate, 40 mM calcium chloride, 10 mg l^-1 adenine sulfate, 0.5% myo-inositol and 6% sucrose gave sustained protoplast divisions. Under these conditions, plating efficiency ranged from 5% for the cultivar Lukulus to 15% for the cultivar Golden Sunrise.Abbreviations BA benzylaminopurine - CaCl₂ calcium chloride, 2,4,-D-2,4-dichlorophenoxyacetic acid - EDTA ethylene diamine tetraacetic acid - KCl potassium chloride - MES-2-N morpholino ethane sulfonic acid - MgCl₂ magnesium chloride - NH₄NO₃ ammonium nitrate - NAA naphthalene acetic acid, p-protoplasts     

Involvement of phosphoenolpyruvate carboxylase and antioxydants enzymes in nitrogen nutrition tolerance in <Emphasis Type="Italic">Sorghum bicolor</Emphasis> plants

Plants of Sorghum bicolor (C₄ species) were grown at different nitrate or ammonium concentrations (0.5, 5, 20 and 50 mM) in order to examine the effect of nitrogen nutrition on growth, phosphoenolpyruvate carboxylase (PEPC) and antioxidant enzymes activities in both roots and leaves of 30-day-old plants. At high NO₃^? levels (20 and 50 mM) the fresh weight was significantly higher. When the nitrogen source was in ammonium form, the leaf and root mass increased drastically at low concentration 5 mM and significantly at 20 mM, however similar fresh weight was found at high level of ammonium (50 mM). The leaves catalase (CAT), guaiacol peroxidase (POD), glutathione reductase (GR), and glutathione S-transferase (GST) activities and the roots glutathione reductase and glutathione S-transferase activities were significantly higher in the NH₄⁺-fed plants than those grown in the nitrate medium. Activity and proteins levels of phosphoenolpyruvate carboxylase in both leaves and roots of sorghum plants were increased progressively with increasing external nitrogen concentration. This increase was more pronounced at high level of ammonium (50 mM), being 2-fold at 50 mM of NO₃^? and 3-fold at 50 mM of NH₄⁺. Our results suggested that antioxidant enzymes activities and PEPC play a key role in ammonium detoxification and tolerance in sorghum plants.