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.     

γ‐Aminobutyric acid addition alleviates ammonium toxicity by limiting ammonium accumulation in rice (Oryza sativa) seedlings

Excessive use of nitrogen (N) fertilizer has increased ammonium (NH₄⁺) accumulation in many paddy soils to levels that reduce rice vegetative biomass and yield. Based on studies of NH₄⁺ toxicity in rice (Oryza sativa, Nanjing 44) seedlings cultured in agar medium, we found that NH₄⁺ concentrations above 0.75 mM inhibited the growth of rice and caused NH₄⁺ accumulation in both shoots and roots. Use of excessive NH₄⁺ also induced rhizosphere acidification and inhibited the absorption of K, Ca, Mg, Fe and Zn in rice seedlings. Under excessive NH₄⁺ conditions, exogenous γ‐aminobutyric acid (GABA) treatment limited NH₄⁺ accumulation in rice seedlings, reduced NH₄⁺ toxicity symptoms and promoted plant growth. GABA addition also reduced rhizosphere acidification and alleviated the inhibition of Ca, Mg, Fe and Zn absorption caused by excessive NH₄⁺. Furthermore, we found that the activity of glutamine synthetase/NADH‐glutamate synthase (GS; EC 6.3.1.2/NADH‐GOGAT; EC1.4.1.14) in root increased gradually as the NH₄⁺ concentration increased. However, when the concentration of NH₄⁺ is more than 3 mM, GABA treatment inhibited NH₄⁺‐induced increases in GS/NADH‐GOGAT activity. The inhibition of ammonium assimilation may restore the elongation of seminal rice roots repressed by high NH₄⁺. These results suggest that mitigation of ammonium accumulation and assimilation is essential for GABA‐dependent alleviation of ammonium toxicity in rice seedlings.     

Ammonium under solution culture alleviates aluminum toxicity in rice and reduces aluminum accumulation in roots compared with nitrate

Al stress and ammonium–nitrogen nutrition often coexist in acidic soils due to their low pH and weak nitrification ability. Rice is the most Al-resistant species among small grain cereal crops and prefers NH₄ ⁺ as its major inorganic nitrogen source. This study investigates the effects of NH₄ ⁺ and NO₃ ^? on Al toxicity and Al accumulation in rice, and thereby associates rice Al resistance with its NH₄ ⁺ preference. Two rice subspecies, indica cv. Yangdao6 and japonica cv. Wuyunjing7, were used in this study. After treatment with or without Al under conditions of varying NH₄ ⁺ and NO₃ ^? supply, rice seedlings were harvested for the determination of root elongation, callose content, biomass, Al concentration and medium pH. The results indicated that Wuyunjing7 was more Al-resistant and NH₄ ⁺-preferring than Yangdao6. NH₄ ⁺ alleviated Al toxicity in two cultivars compared with NO₃ ^?. Both NH₄ ⁺-Al supply and pretreatment with NH₄ ⁺ reduced Al accumulation in roots and root tips compared with NO₃ ^?. NH₄ ⁺ decreased but NO₃ ^? increased the medium pH, and root tips accumulated more Al with a pH increase from 3.5 to 5.5. Increasing the NO₃ ^? concentration enhanced Al accumulation in root tips but increasing the NH₄ ⁺ concentration had the opposite effect. These results show NH₄ ⁺ alleviates Al toxicity for rice and reduces Al accumulation in roots compared with NO₃ ^?, possibly through medium pH changes and ionic competitive effects. Making use of the protective effect of NH₄ ⁺, in which the Al resistance increases, is advised for acidic soils, and the hypothesis that rice Al resistance is associated with the preferred utilization of NH₄ ⁺ is suggested.     

Ammonium-Stimulated Root Hair Branching is Enhanced by Methyl Jasmonate and Suppressed by Ethylene in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Root hair development is orchestrated by nutritional factors and plant hormones. We investigated the action of ammonium (NH₄⁺) and its interactions with methyl jasmonate (MeJA) and ethylene in Arabidopsis root hair growth. The formation of root hair branches was dramatically stimulated in media containing 1.25 to 20 mM NH₄⁺ at pH values of 4.0 to 6.5. The NH₄⁺-treated root hairs showed a very short tip growth stage and swells on the sides that indicated the emergence of branches. MeJA (0.08 to 10 μM) worked in synergism with NH₄⁺ to enhance hair branching. In contrast, ethylene had an antagonistic effect; the stimulation of hair branching by NH₄⁺ was suppressed by the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) and was diminished in ethylene-overproducing mutant eto1-1 seedlings. Moreover, the application of Ag⁺, an ethylene inhibitor, reduced the ACC-induced inhibition of NH₄⁺-stimulated hair branching and restored NH₄⁺-stimulated hair branching in eto1-1 seedlings. Thus, the actions of jasmonate and ethylene appear to be dependent on nutritional conditions such as available nitrogen.     

Root growth inhibition by NH4+ in Arabidopsis is mediated by the root tip and is linked to NH4+ efflux and GMPase activity

Root growth in higher plants is sensitive to excess ammonium (NH₄⁺). Our study shows that contact of NH₄⁺ with the primary root tip is both necessary and sufficient to the development of arrested root growth under NH₄⁺ nutrition in Arabidopsis. We show that cell elongation and not cell division is the principal target in the NH₄⁺ inhibition of primary root growth. Mutant and expression analyses using DR5:GUS revealed that the growth inhibition is furthermore independent of auxin and ethylene signalling. NH₄⁺ fluxes along the primary root, measured using the Scanning Ion‐selective Electrode Technique, revealed a significant stimulation of NH₄⁺ efflux at the elongation zone following treatment with elevated NH₄⁺, coincident with the inhibition of root elongation. Stimulation of NH₄⁺ efflux and inhibition of cell expansion were significantly more pronounced in the NH₄⁺‐hypersensitive mutant vtc1‐1, deficient in the enzyme GDP‐mannose pyrophosphorylase (GMPase). We conclude that both restricted transmembrane NH₄⁺ fluxes and proper functioning of GMPase in roots are critical to minimizing the severity of the NH₄⁺ toxicity response in Arabidopsis.     

Nitrogen use efficiency (NUE) in rice links to NH4 + toxicity and futile NH4 + cycling in roots

Aims

Rice is known as an ammonium (NH₄ ⁺)-tolerant species. Nevertheless, rice can suffer NH₄ ⁺ toxicity, and excessive use of nitrogen (N) fertilizer has raised NH₄ ⁺ in many paddy soils to levels that reduce vegetative biomass and yield. Examining whether thresholds of NH₄ ⁺ toxicity in rice are related to nitrogen-use efficiency (NUE) is the aim of this study.

Methods

A high-NUE (Wuyunjing 23, W23) and a low-NUE (Guidan 4, GD) rice cultivar were cultivated hydroponically, and growth, root morphology, total N and NH₄ ⁺ concentration, root oxygen consumption, and transmembrane NH₄ ⁺ fluxes in the root meristem and elongation zones were determined.

Results

We show that W23 possesses greater capacity to resist NH₄ ⁺ toxicity, while GD is more susceptible. We furthermore show that tissue NH₄ ⁺ accumulation and futile NH₄ ⁺ cycling across the root-cell plasma membrane, previously linked to inhibited plant development under elevated NH₄ ⁺, are more pronounced in GD. NH₄ ⁺ efflux in the root elongation zone, measured by SIET, was nearly sevenfold greater in GD than in W23, and this was coupled to strongly stimulated root respiration. In both cultivars, root growth was affected more severely by high NH₄ ⁺ than shoot growth. High NH₄ ⁺ mainly inhibited the development of total root length and root area, while the formation of lateral roots was unaffected.

Conclusions

It is concluded that the larger degree of seedling growth inhibition in low- vs. high-NUE rice genotypes is associated with significantly enhanced NH₄ ⁺ cycling and tissue accumulation in the elongation zone of the root.     

Shoot-supplied ammonium targets the root auxin influx carrier AUX1 and inhibits lateral root emergence in Arabidopsis

Deposition of ammonium (NH₄⁺) from the atmosphere is a substantial environmental problem. While toxicity resulting from root exposure to NH₄⁺ is well studied, little is known about how shoot‐supplied ammonium (SSA) affects root growth. In this study, we show that SSA significantly affects lateral root (LR) development. We show that SSA inhibits lateral root primordium (LRP) emergence, but not LRP initiation, resulting in significantly impaired LR number. We show that the inhibition is independent of abscisic acid (ABA) signalling and sucrose uptake in shoots but relates to the auxin response in roots. Expression analyses of an auxin‐responsive reporter, DR5:GUS, and direct assays of auxin transport demonstrated that SSA inhibits root acropetal (rootward) auxin transport while not affecting basipetal (shootward) transport or auxin sensitivity of root cells. Mutant analyses indicated that the auxin influx carrier AUX1, but not the auxin efflux carriers PIN‐FORMED (PIN)1 or PIN2, is required for this inhibition of LRP emergence and the observed auxin response. We found that AUX1 expression was modulated by SSA in vascular tissues rather than LR cap cells in roots. Taken together, our results suggest that SSA inhibits LRP emergence in Arabidopsis by interfering with AUX1‐dependent auxin transport from shoot to root.     

Effect of nitrogen form and phosphorus source on the growth,nutrient uptake and rhizosphere soil property of Camellia sinensis L.

The effects of nitrogen form and phosphorus source on the growth, nutrient uptake and rhizosphere soil property of tea (Camellia sinensis L.) were investigated in a pot experiment. The experiment was performed with a compartmental cropping device, which enables the collection of rhizosphere soil at defined distances from the root of tea plant. Nitrogen was supplied as nitrate or ammonium in combination with soluble phosphorus as Ca(H₂PO₄)₂ or insoluble P as rock phosphate. The leaf dry matter production of tea was significantly greater in the treatments with NH₄ ⁺ than NO₃ ^-, whereas dry matter production of root and stem was not significantly affected. Addition of phosphorus as either source did not influence the dry matter production. The concentrations of K in root, Mg and Ca in both the shoot and root supplied with NO₃ ^- were significantly higher than in NH₄ ⁺ and influence of P sources was minor. On the contrary, Al and Mn concentrations were significantly larger in NH₄ ^--fed plants which could be attributed to remarkably increased availability of Al and Mn caused by acidification of the rhizosphere soil (the first 1-mm soil section from the root surface) with NH₄–N nutrition. The concentration of N in shoot was also significantly higher in NH₄- than in NO₃-fed plants, indicating higher use efficiency of NH₄–N. Whatever the phosphate source, rhizosphere pH declined in ammonium compared to in nitrate treatment. The pH decrease was much larger when no P or soluble P were applied and reached 0.85–1.30 units which extended to 3–5 mm away from the root surface. Exchangeable acidity, content of exchangeable Al and Mn were also considerably higher in the rhizosphere soils of NH₄ ⁺ fed tea plants. Significant amounts of P dissolved from rock phosphate accumulated in rhizosphere of NH₄ ⁺, not NO₃ ^-, suggesting that the dissolution of rock phosphate was induced by the proton excreted by tea root fed with ammonium. With soluble P addition, shoot and root P concentrations were greater in NH₄ ⁺ than in NO₃ ^- treatment and it appeared that this difference could not be sufficiently explained by the available P content in soil which was only slightly higher in NH₄ ⁺ treatment. With rock phosphate addition, the shoot and root P concentrations were hardly affected by nitrogen form, although the available P content was much higher and accumulated in the rhizosphere soil supplied with ammonium. The reason for this was discussed with regard to the inter-relationship of Al with P uptake. This revised version was published online in June 2006 with corrections to the Cover Date.     

Absorption of nitrate and ammonium ions by Lolium perenne from flowing solution cultures at low root temperatures

Lolium perenne L. cv. 23 (perennial ryegrass) plants were grown in flowing solution culture and acclimatized over 49 d to low root temperature (5°C) prior to treatment at root temperatures of 3, 5, 7 and 9°C for 41 d with common air temperature of 20/15°C day/night and solution pH 5·0. The effects of root temperature on growth, uptake and assimilation of N were compared with N supplied as either NH₄ or NO3 at 10 mmol m^?3. At any given temperature, the relative growth rate (RGR) of roots exceeded that of shoots, thus the root fraction (Rf) increased with time. These effects were found in plants grown with the two N sources. Plants grown at 3 and 5°C had very high dry matter contents as reflected by the fresh weight: freeze-dried weight ratio. This ratio increased sharply, especially in roots at 7 and 9°C. Expressed on a fresh weight basis, there was no major effect of root temperature on the [N] of plants receiving NHJ but at any given temperature, the [N] in plants grown with NHJ was significantly greater than in those grown with NO₃. The specific absorption rate (SAR) of NH⁺₄ was greater at all temperatures than SAR-NO₃. In plants grown with NH⁺, 3–5% of the total N was recovered as NH⁺₄, whereas in those grown with NO^?₃ the unassimilated NO^?₃ rose sharply between 7 and 9°C to become 14 and 28% of the total N in shoots and roots, respectively. The greater assimilation of NH⁺₄ lead to concentrations of insoluble reduced N (= protein) which were 125 and 20% greater, in roots and shoots, respectively, than in NO^?₃-grown plants. Plants grown with NH⁺₄ had very much greater glutamine and asparagine concentrations in both roots and shoots, although other amino acids were more similar in Concentration to those in NO^?₃ grown plants. It is concluded that slow growth at low root temperature is not caused by restriction of the absorption or assimilation of either NH⁺₄ or NO^?₃. The additional residual N (protein) in NH⁺₄ grown plants may serve as a labile store of N which could support growth when external N supply becomes deficient.     

Nitrate reductase of primary roots of red spruce seedlings : effects of acidity and metal ions

Nitrate reductase activity (NRA) was found in primary roots, but not in foliage of red spruce (Picea rubens Sarg.) seedlings. Nitrate induced NRA:NH₄⁺ did not induce and slightly depressed NRA in older seedlings. Induction required 8 hours and, once induced, NRA decreased slowly in the absence of exogenous NO₃⁻. Seedlings were grown in perlite with a complete nutrient solution containing NH₄⁺ to limit NR induction. Established seedlings were stressed with nutrient solutions at pH 3, 4, or 5 supplemented with Cl⁻ salts of Al, Cd, Pb, or Zn each at two concentrations. NRA in primary root tips was measured at 2, 14, 28, and 42 days. NRA induction was greatest at pH 3, and remained high during the period of study. NRA induction at pH 4 was lower. Metal ions suppressed NRA at pH 3 and 5, but enhanced NRA at pH 4. It is concluded that acidity and soluble metals in the root environment of red spruce are unlikely to be important factors in nitrogen transformations in red spruce roots.     

The effect of root temperature on the uptake of nitrogen and the relative size of the root system in Lolium perenne. I. Solutions containing both NH+4 and NO3−

Abstract Lolium perenne L. cv. S23 was grown in flowing culture solution, pH 5, in which the concentrations of NH₄⁺, NO₃^? and K⁺ were frequently monitored and adjusted to set values. In a pre-experimental period, plants were acclimatized to a regime in which roots were treated at 5°C with shoots at 25°C. The root temperature was then changed to one of the following, 3, 7, 9, 11, 13, 17 or 25°C, while air temperature remained at 25°C. When root temperature was increased from 5X, the relative growth rate of roots increased immediately while that of shoots changed much less for a period of approximately 9 d (phase 1). Thus, the root: shoot ratio increased, but eventually approached a new, temperature-dependent, steady value (phase 2). The fresh: freeze-dried weight ratio (i.e. water content) in shoots (and roots) increased during the first phase of morphological adjustment (phase 1). In both growth phases and at all temperatures, plants absorbed more NH₄⁺ than NO₄⁺, the tendency being extreme at temperatures below 9° where more than 85% of the N absorbed was NH₄⁺. Plants at different root temperatures, growing at markedly different rates, had very similar concentrations of total N in their tissues (cells) on a fresh weight basis, despite the fact that they derived their N with differing preference for NH₄⁺. Specific absorption rates for NH₄⁺, NO_x^?, K⁺ and H₂PO₄^? showed very marked dependence on root temperature in phase 1, but ceased to show this dependence once a steady state root: shoot ratio had been established in phase 2. The results indicate the importance of relative root size in determining ion fluxes at the root surface. At higher temperatures where the root system was relatively large, ‘demand’ per unit root was low, whereas at low temperatures roots were small relative to shoots and ‘demand’ was high enough to offset the inhibitory effects of low temperature on transport processes.     

Effects of soil pH and nitrogen fertility on the population dynamics of Thielaviopsis basicola

Black root rot of tobacco, caused by Thielaviopsis basicola, is generally severe at soil pH values >5.6 and suppressed under more acidic conditions (pH < 5.2). Soil acidifying fertilizers containing NH₄–N are generally recommended for burley tobacco production in North Carolina, but the effects of N form and application rate on development of black root rot and on the population dynamics of T. basicola have not been determined. Greenhouse and laboratory studies were conducted to evaluate the effects of N form (NH₄ ⁺ or NO₃ ^–) and rate on pathogen and disease parameters at several initial soil pH levels. A moderately-conducive field soil, initial pH 4.7, was adjusted to a pH of 5.5 or 6.5 by the addition of CaOH₂, then amended with the desired nitrogen form and rate. Pathogen populations were determined over time. In addition, spore production in extracts of roots from plants grown in the various nitrogen and pH treatments was determined. Finally, because tobacco responds to acidic soil conditions and exposure to NH₄–N by accumulating high concentrations of the polyamine putrescine, the toxicity of putrescine on vegetative growth and reproduction of T. basicola was investigated. Low soil pH and high levels of NH₄–N suppressed reproduction of T. basicola in soil and in root extract, while use of NO₃–N and depletion of NH₄–N resulted in rapid increases in populations of T. basicola. At 20 mM, putrescine inhibited hyphal growth by 60% and aleuriospore production by 98%. Fertilizers that reduced soil pH also reduced reproduction by T. basicola, and thus have potential for management of black root rot by suppressing populations of T. basicola over multiple years of crop production. The suppression of T. basicola and black root rot observed with NH₄–N amendments may partially be due to development of an inhibitory environment in the root and not solely to changes in rhizosphere pH.     

Accumulation of ammonium ion in cadmium tolerant and sensitive cultivars of Oryza sativa

Cd-tolerant and Cd-sensitive rice cultivars were used to study the role of NH₄ ⁺ accumulation in Cd-induced toxicity. NH₄ ⁺ accumulation seems to be involved in regulating the toxicity of rice seedlings caused by CdCl₂. This conclusion was based on the observations that (a) on treatment with CdCl₂, NH₄ ⁺ content increased rapidly in the leaves of the Cd-sensitive cultivar (cv. Taichung Native 1, TN1) but not in the Cd-tolerant cultivar (cv. Tainumg 67, TNG67), (b) pretreatment with abscisic acid (ABA) enhanced Cd tolerance and reduced Cd-induced NH₄ ⁺ accumulation in TN1 seedlings, (c) exogenous application of the ABA biosynthesis inhibitor, fluridone, decreased Cd tolerance and increased NH₄ ⁺ content in leaves of TNG67, (d) exogenous application of phosphinothricin, an inhibitor of glutamine synthetase (GS), which resulted in NH₄ ⁺ accumulation in the leaves, also induced toxicity similar to Cd in TN1 seedlings. Evidence is presented to show that Cd-induced NH₄ ⁺ accumulation in TN1 leaves is attributable to a decrease in GS activity. Since Cd-treated TN1 leaves had higher glutamine and glutamate contents than control leaves, it is unlikely that glutamine (or glutamate) depletion is the mechanism which regulates Cd-induced toxicity.     

Seedling responses to band applied NH4OH rates and to N form in two maize hybrids

Growth of maize seedlings can be improved by enhanced ammonium nutrition, but placing fertilizer anhydrous ammonia close to seedlings introduces the risk of ammonia toxicity. In this study, growth and root elongation response to rates of closely placed NH₄OH bands were investigated in two contrasting maize hybrids. Seven rates of NH₄OH, ranging from 0 to 200 mg N kg^-1 soil were injected into the center of each pot. A single rate of Ca(NO₃)₂-N was included to compare hybrids for N form preference at a moderate N rate. Three seedlings per pot were planted 5.7 cm from the injection point.Hybrid B73×LH51 produced a quadratic response in shoot growth to NH₄OH rates, whereas LH74×LH123 exhibited a significant linear decline in response to NH₄OH rate. Root length density sampled from the fertlized zone declined linearly in response to NH₄OH rate while a slight increase in root length density in unfertilized zones was observed at intermediate NH₄OH rates. Hybrids did not differ in root length density in either zone.The hybrid with greater tolerance of NH₄OH rates (B73×LH51) also showed a preference in shoot growth for NH₄-over NO₃-N at 66.7 mg N kg^-1 compared to LH74×LH123. On average across hybrids, nitrate concentrations of xylem exudate collected from detopped plants were 14.5 mmol g^-1 for Ca(NO₃)₂ treatments and 1.5 mmol g^-1 for NH₄OH treatments, indicating that contrasting N-form nutrition resulted from fertilizer treatments. Malate concentrations were higher in the NH₄OH treatment indicating that this organic acid anion may substitute for the negative charge of nitrate during enhanced ammonium nutrition in maize.The results suggest that potentially useful genetic variation exists in maize for N form preference and for tolerance to increasing ammonical-N rates.     

Effect of exogenous ammonium gluconate on growth,ion flux and antioxidant enzymes of maize (Zea Mays L.) seedlings under NaCl stress

• Ammonium gluconate (AG) provides both an organic carbon source and a nitrogen source, which can positively improve soil fertility and delay soil degradation.
• We investigated the underlying mechanisms of both NH₄⁺‐ and C₆H₁₁O₇^?‐mediated resistance to high salt concentrations in maize (Zea mays L.), and how they relate to antioxidant cellular machinery, root system architecture, root activity and lignin content in roots.
• Seedlings treated with AG maintained lower Na⁺ content, higher chlorophyll content, higher CAT and POD activity, compared with those without AG and ammonium carbonate (AC). The total size of the root system, primary root length and number of lateral roots detected on the primary root treated with AG decreased compared with those not treated with AG at the same NaCl concentration. However, average root diameter and root activity when treated with AG were significantly higher than roots without AG at the same NaCl concentration. Furthermore, total size of the root system, primary root length and number of lateral roots detected on primary rootsof seedlings treated with AG were higher than those treated with AC at the same NaCl concentration.
• These results suggested that AG may be a good organic fertiliser under salt stress by decreasing Na⁺ content and increasing chlorophyll content, activity of antioxidant enzymes, root diameter and root activity in maize seedlings.

     

Regulation of NH4 + transport pathways in Pisum arvense roots

The effects of metabolic and protein synthesis inhibitors on NH₄ ⁺ uptake by Pisum arvense plants at low (0.05 mM) and high (1 mM) external ammonium concentration were studied. In short-time experiments cycloheximide decreased the ammonium uptake rate at low level of NH₄ ⁺ and increased the absorption of NH₄ ⁺ from uptake medium containing high ammonium concentration. Arsenate and azide supplied into uptake solutions at low ammonium concentration strongly decreased or completely suppressed the NH₄ ⁺ uptake rate, respectively. When the experiments were carried out at high level of ammonium only azide decreased the uptake rate of NH₄ ⁺ and arsenate stimulated this process. Dinitrophenol very strongly repressed the uptake rate of NH₄ ⁺ at both ammonium concentrations. After removing dinitrophenol from both solutions, neither at low nor high external ammonium level the recovery of NH₄ ⁺ uptake rate was achieved within 150 min or 3 h, respectively. The recovery of NH₄ ⁺ uptake rate after removing azide was observed within 90 min and 3 h at low and high ammonium concentrations, respectively. The regulation of NH₄ ⁺ uptake by some inhibitors at low external ammonium level was investigated using plasma membrane vesicles isolated from roots by two-phase partitioning. Orthovanadate completely suppressed the uptake of NH₄ ⁺ by vesicles and quinacrine decreased the NH₄ ⁺ uptake which 55 suggests that ammonium uptake depends on activities of plasma membrane-bound enzymes. On the other hand, it was found that dinitrophenol completely reduced the NH₄ ⁺ uptake by vesicles. The various effects of inhibitors on ammonium uptake dependent on external ammonium concentration suggest the action of different ammonium transport systems in Pisum arvense roots. The ammonium transport into root cells at low NH₄ ⁺ level requires energy and synthesis of protein in the cytoplasm. The research was supported by grant of KBN No. 6PO4C 068 08     

Effect of nitrogen forms on growth and chemical changes in the rhizosphere of strawberry plants

The experiment was set up to examine the influence of different nitrogen forms: (NH₄)₂SO₄, Ca(NO₃)₂ or NH₄NO₃ on growth response, root induced pH changes in the rhizosphere, root-borne acid phosphatase activity in strawberry plants cv. Senga Sengana. The plants grown on sandy mineral soil were fertilized with 3 forms of nitrogen, in concentrations of 46 mg N·kg⁻¹ soil. The plants were grown in rhizoboxes with removable plexiglass lids. To ensure the root growth along the plexiglass lids, the rhizoboxes were placed at an angle of about 50° with the lid on the lower side. In case of ammonium supply, the nitrification inhibitor DIDIN was added (10 mg·kg⁻¹ of moist soil) to prevent conversion of ammonium into nitrate. The growth response (roots and shoots) of strawberry plants were determined after 11 weeks of treatment with different N forms. The best development of the root system and shoots (root and shoot dry weight and root length) was obtained, when ammonium nitrate was supplied. It is suggested therefore, that NH₄NO₃ stimulates vegetative growth of strawberry plants cv. Senga Sengana. However, there were no statistical differences in a leaf and flower number of the plants grown under different forms of N-fertilization. Determination of rhizosphere pH, and acid phosphatase activity were executed using non-destructive techniques, which enabled weekly measurement of chemical changes in the rhizosphere. The results revealed that the form of nitrogen supplied had a predominant effect on chemical changes in the rhizosphere of strawberry plants. The highest pH values (average pH 6.8) were measured in the rhizosphere of individual plants supplied with Ca(NO₃)₂. Whereas the lowest pH values (average pH 5.8) were detected in the presence of (NH₄)₂SO₄. The curve of rhizosphere pH measured along individual roots of the plants treated with Ca(NO₃)₂ represents the highest pH values whereas the curve of rhizosphere pH under (NH₄)₂SO₄ treatment had the lowest pH values. The highest activity of acid phosphatase were observed in the rhizosphere of strawberry plants grown in the presence of (NH₄)₂SO₄, at pH 5.8.     

Effects of different nitrogen forms on the growth and cytokinin content in xylem sap of tomato (Lycopersicon esculentum Mill.) seedlings

In order to investigate the effects of homogeneous and localized supply of different nitrogen forms (nitrate, NO₃ ^? vs ammonium, NH₄ ⁺) on the growth of tomato seedlings, root morphology and six cytokinin (CTK) fractions in xylem sap were analyzed. Whole roots were supplied with different ratios of NO₃ ^? to NH₄ ⁺ (100:0, as 100-0NA; 75:25, as 75-25NA; 50:50, as 50-50NA) under homogeneous supply. In split-root experiments, three treatments were compared: a sole NO₃ ^? supply (N|N), a spatially separated supply of NO₃ ^? and NH₄ ⁺ (N|A), and a spatially separated supply of NO₃ ^? and a mixture of NO₃ ^? and NH₄ ⁺ nutrition at a ratio of 75:25 (N|AN). All concentrations of total N were set at 5 mM. The results showed that (1) homogeneous 75% NO₃ ^? plus 25% NH₄ ⁺ supply to the whole root zone led to maximum shoot and root dry matter (DM), root surface area (RS) and root volume (RV). The spatially separated supply of NO₃ ^? and NH₄ ⁺ (N|A) resulted in a contrasting effect on root morphology: in comparison to N|N, root DM in the NO₃ ^?-containing pot was increased by 50% whereas it was depressed by 50% in the NH₄ ⁺-containing pot. The 75% NO₃ ^? plus 25% NH₄ ⁺ supply in the split-root experiment led to no significant effects either on shoot DM and root DM, or on RS and RV when compared to N|N. (2) The presence of NH₄ ⁺ in the external medium led to a significantly reduced total xylem-CTK concentration, and a close negative correlation was found between xylem NH₄ ⁺ and total CTK concentration irrespective of culture mode. A relatively high level of zeatin riboside (ZR) was maintained both in 75-25NA and N|A treatments. It was concluded that, in addition to the percentage of NH₄ ⁺ to NO₃ ^? in the nutrient solution, whether NH₄ ⁺ was supplied to the whole root system or to only part of the root system was also an important factor affecting plant growth. The fact that the 75-25NA and N|A treatments resulted in optimal growth of tomato seedlings might be attributed to the higher ZR concentration in xylem.     

Response of Douglas fir seedlings to nitrate and ammonium nitrogen sources at different levels of pH and iron supply

Douglas fir seedlings were grown for two to three months in sand and soil cultures in a greenhouse to examine their growth response to nitrogen (N) source at different levels of pH and iron (Fe) supply. In the first two experiments nutrient solutions of known pH were automatically applied to the top of the sand cultures and allowed to run to waste from the bottom. Under these conditions seedlings made most growth on nitrate (NO₃–N) under acid (pH4) conditions, but most growth on ammonium (NH₄–N) under neutral (pH7) conditions. Calcium carbonate (CaCO₃) was used to create a range of pH conditions (from 4.0 to 7.2) in a peat and sand artificial soil. Over the pH range 4 to 6 NH₄–N or NO₃+NH₄–N produced larger seedlings than NO₃–N alone, but above pH6 growth on all N sources was depressed. Chemical analysis showed that seedling Ca concentration had increased and Fe concentration had decreased with increase in CaCO₃ application. Both Ca and Fe concentrations were higher in seedlings receiving NO₃–N than in those receiving NH₄ or NO₃+NH₄.In sub-irrigated sand cultures, Doughlas fir seedlings receiving NO₃–N were shown to respond to additions of Fe chelate, but seedlings receiving NH₄–N responded little to Fe chelate. At pH5 seedlings receiving NO₃–N did not grow as big as seedlings receiving NH₄–N in the absence of Fe chelate, but addition of Fe chelate resulted in NO₃-fed seedlings growing larger than NH₄-fed seedlings. The relationship between seedling Fe concentration and N nutrition is discussed.The relatively larger root dry weight and surface area of seedlings grown on NO₃–N, as compared to NH₄–N, in sand culture, was noted.     

生长素和乙烯互作调控硝酸铵诱导的根毛分叉

以野生型拟南芥(WT)及其生长素和乙烯不敏感型突变体(aux1-7、axr1-3、etr1-1和etr1-3)为实验材料,采用固体培养法研究了高浓度硝酸铵对根毛发育的影响,以揭示其调控根毛发育的机制。结果表明:(1)随着外源硝酸铵浓度的逐渐增加,拟南芥根毛伸长受阻,产生大量的分叉根毛。(2)高浓度硝酸铵条件下,外源活性氧或活性氧产生抑制剂二苯基氯化碘(DPI)的添加能抑制高浓度硝酸铵诱导的分叉根毛产生。(3)高浓度硝酸铵条件下,外源生长素或乙烯合成前体物质1-氨基-环丙烷-1-羧酸(ACC)处理能恢复根毛的正常生长,解除高浓度硝酸铵诱导根毛分叉现象。(4)高浓度硝酸铵条件下,外源生长素处理乙烯不敏感型突变体或ACC处理生长素不敏感型突变体均能抑制突变体分叉根毛的形成。研究表明,活性氧、生长素和乙烯都参与了高浓度硝酸铵对根毛发育的过程调控;在硝酸铵诱导的根毛分叉中生长素和乙烯存在相互作用,在缺乏生长素信号通路时,乙烯能够发挥补充作用抑制分叉根毛的产生;在缺乏乙烯信号通路时,生长素也可以弥补缺失乙烯的作用抑制根毛的分叉,但是需要更高浓度的生长素才能充分抑制分叉根毛的产生。