Intraspecific differences in nitrogen assimilating enzymes in spinach under contrasting forms of nitrogen supply 1

Intraspecific differences in the activities of nitrate reductase (NR), glutamine synthetase (GS), NADH dependent glutamate synthase (NADH‐GOGAT), and glutamate dehydrogenase (GDH) under contrasting forms of nitrogen (N) supply were studied in tissues of three spinach (Spinacia oleracea L.) cultivars. The varieties (Viroflay, Butterflay, and Giant) were smooth, curly and semicurly leaved, respectively. The plants were grown in nutrient solutions containing NO₃ as the sole source of N (100:0) and NO₃ plus NH₄ (80:20). Giant, the NH₄ tolerance of which had been evaluated in growth and on the basis of nutrient uptake, had much higher GS and GDH activities in the roots and higher NR and NADH‐GOGAT levels in the leaves of plants grown on NO₃ and NH₄ than that grown on NO₃ alone. On the level of N assimilating enzymes of Butterflay, mixed N nutrition caused an increase of GDH and NADH‐GOGAT in leaves and roots and at the same time a decrease of GS in the roots and NR in the leaves. An inverse relationship between GS and GDH activities was detected in the leaves and foots of Virofiay grown with both N sources. Finally, Viroflay gave the highest levels of GDH irrespective of the NO₃:NH₄ assayed, whereas the leaves of Giant were GDH deficient in comparison with the other cultivars. In addition, the GS activity approached zero in the roots of spinach cultivars characterized by hardly any NH₄ tolerance, whereas in those of Giant it increased remarkably with the supply of NO₃ plus NH₄.     

Nitrogen Metabolism in Tomato Plants Under Cadmium Stress

Abstract

Young seedlings of tomato (Lycopersicon esculentum Mill.) grown on nitric medium and treated with different cadmium (Cd) concentrations (0–50 µM) were used. Results obtained show that Cd remains predominantly located in the roots, which then seem to play the role of trap‐organs. Increasing Cd concentration in the medium leads particularly to a decrease in NO₃ ^? accumulation, together with decrease in the activities of nitrate reductase (NR), nitrite reductase (NiR), and glutamine synthetase (GS). On the other hand, stimulations were observed for ammonium accumulation, anabolitic activity of glutamate dehydrogenase (GDH) and protease activity. Contrarily, decreases were obtained for soluble protein contents and catabolitic activity of GDH. These results were discussed in relation to the hypothesis attributing to the mitochondrial enzyme NADH‐dependent GDH, an important role in the plant defense processes, in maintaining the glutamate pool in conditions where the need for NH₄ ⁺ detoxification is increased by heavy metals.     

Influence of plant growth regulators on some enzymes of nitrogen assimilation in mustard seedlings

Effect of plant growth regulators (PGR) viz. gibberellic acid (GA), kinetin (KN), and abscissic acid (ABA) were investigated on growth and activities of nitrate reductase (NR) and glutamine synthetase (GS) in mustard (Brassica juncea) seedlings. All the PGRs tested promoted in vivo NR activity in cotyledons, but the magnitude differed with different treatments. Cytosolic GS in root and hypocotyl was promoted by GA treatment and inhibited by ABA and KN treatments although the latter showed slight promotion initially in hypocotyl; the trend was not clear in cotyledons. Determination of K_m value of GS extracted from 96‐h‐old cotyledons recorded lower K_m value in GA treatment (2.5 mM), while it increased in ABA treatment (4.35 mM), There was little change in K_m value in KN (3.03 mM) treatment. The kinetics of GS enzyme in cotyledons of different treatments showed marked variation in V_max . Both GA and ABA treatments inhibited GS activity while no significant effect by KN treatment was observed. It is argued that GA treatment inhibits chloroplastic GS (an enzyme which has higher K_m value), while ABA‐induced inhibition may not be specific to cytosolic or chloroplastic isoforms. Kinetin treatment was ineffective in promoting or inhibiting GS activity in cotyledons. The above conclusion is further supported by chloroplastic pigment data where inhibition is recorded in all the PGRs tested.     

The role of root exudates in specific apple (Malus × domestica Borkh.) replant disease (SARD)

In an effort to improve our understanding of the specific apple replant disease (SARD), direct and indirect effects of phytohormones and related compounds (abscisic acid, 6‐benzyladenine, indole‐3‐acetic acid, 1‐naphthaleneacetic acid, and gibberellins GA₃ or GA₄) on root exudates of apple seedlings were evaluated as a potential mediating factor in the infection process. In the first type of experiments, radioactively labeled hormonal substances were applied to the stumps of decapitated apple‐seedlings and the occurrence of radioactivity in root exudates and their qualitative characterization were examined (direct influence). In another set of experiments, the effects of leaf‐applied plant growth regulators on the amount and composition of the predominant organic acids, carbohydrates, and amino acids/amides were studied. Cherry seedlings resistant to SARD and apple seedlings with dormant apical buds and, thus, not susceptible to infection were used for comparison. The results showed no differences in exudation of applied plant growth regulators between growing cherry and apple seedlings. Thus, a direct effect of plant hormones on the infection process is unlikely. However, leaf treatments with growth regulators, in particular with auxin‐type compounds and abscisic acid, increased exudation of alditols. This may indicate that plant hormones are, to some degree, indirectly involved in the infection process.     

Glutamate dehydrogenase in higher plants

The occurrence of NADPH-dependent glutamate dehydrogenase (GDH) was studied in higher plants. All of the higher plants tested had both NADH- and NADPH-dependent GDH activities, based on the following observations: 1) NADPHdependent GDH activity was found in the extracts of corn and soybean leaves, which was free of NADPH-dehydrogenase by heat treatment or DEAE-cellulose column chromatography. 2) Radish leaves and roots grown under germ-free condition showed the same NADPH-dependent GDH activity as those grown under conventional condition. 3) No conversion of NADPH to NADH in the reaction mixture was demonstrated by measuring alcohol dehydrogenase activity. Moreover, it was ascertained that rice plant cells grown on suspension culture had 50% of NADPH-dependent GDH activity, and both activities were not affected by nitrogen sources.     

The effects of triacontanol on seedling growth and polyphenol oxidase activity in dark and light growth lettuce

The present study was designed to study the effects of various concentrations (0.00; 0.01, 0.10 and 1.00‐mg/ml) of triacontanol on root, stem and leaf growth and polyphenol oxidase activity in each respective tissue; in Grand Rapids and Great Lakes varieties of eleven‐day‐old dark and light grown lettuce seedlings.

Root, stem and leaf growth was less than in non‐triacontanol‐treated Grand Rapids seedlings in both TC‐treated dark and light exposed seedlings. With respect to Great Lakes seedlings, all dark‐grown roots exhibited greater growth than the corresponding untreated control. Light‐grown Great Lakes roots treated with 0.01‐mg/ml and 1.0‐mg/ml or triacontanol respectively, grew more than control or 0.1‐mg/ml triacontanol‐treated seedlings. Both dark and light‐grown triacontanol‐treated stem and leaf tissues of Great Lakes seedlings all produced less growth than the untreated controls.

The Grand Rapids variety had less polyphenol oxidase activity in both dark and light‐grown root and stem tissues than in untreated controls; however, both dark and light‐grown leaf tissue, treat ed with 0.01‐mg/ml and 1.0‐mg/ml of triacontanol respectively exhibited more polyphenol oxidase activity than 1.0‐mg/ml triacontanol ‐treated or untreated control tissues.

TC treatment of 0.1‐mg/ml caused no enhancement of PPO activity in dark or light‐grown root, stem and leaf tissues of Great Lakes tissue, however, seedlings treated with a concentration of 0.01‐mg/ml TC exhibited more PPO activity than non‐TC‐treated controls.     

外源蔗糖对高NH4+胁迫下拟南芥碳氮代谢的影响

以拟南芥野生型、amt1.1和amt1.3为实验材料，采取土培的方法，以正常培养液（4mmol/L NH4+）培养，在20mmol/L NH4+的胁迫下，通过在培养液中添加0%（T1）蔗糖、5%（T2）蔗糖，测定地上部分的鲜重，叶绿素，游离NH4+，可溶性糖，可溶性蛋白，谷氨酰胺合成酶（GS）、谷氨酸脱氢酶（GDH），矿质元素含量等指标，研究外源蔗糖对NH4+胁迫拟南芥碳氮代谢的影响。结果表明，T1处理下，拟南芥生长受到严重的抑制，鲜重，GS，GDH酶活性降低，游离NH4+含量，叶绿素含量，可溶性糖和可溶性蛋白含量增加，植株的N、P、K、Ca的含量增加，Mg、Fe的含量减少。其中col-0在T1处理下受到的抑制比amt1.1和amt1.3更为显著。与T1处理相比较，T2处理增加了拟南芥植株的鲜重，显著提高了可溶性糖和可溶性蛋白含量，提高了GS和GDH的活性；降低了叶绿素和游离NH4+的含量，提高了植株体内的N、P、K、Ca，Mg的含量，降低了植株Fe的含量，其中，外源蔗糖对col-0高NH4+毒害的缓解更为显著。     

Phytotoxic Effects of Residual Glufosinate on the Nitrogen Assimilation of Transplanted Rapeseed Seedlings

Herbicide application is an efficient method to control weed growth in modern agriculture production, but there is concern about the ecological impact of unwanted herbicide residues in the soil. Rapeseed varieties ZS11 and D148 were used to evaluate the phytotoxic effects of residual glufosinate on the assimilation of nitrogen (N) in rapeseed seedlings transplanted to untreated [0 g hm^?2 glufosinate] or treated [450 g hm^?2 and 900 g hm^?2 glufosinate] soils. Glutamine synthetase (GS) and glutamate dehydrogenase (GDH) activities, the contents of ammonium (NH₄⁺), free amino acids (FAA), and soluble protein (SP), and seedling dry weight (DW) were determined at 5, 8, 11, 20, 40, and 70 d post-transplant. Both concentrations of glufosinate induced physiological phytotoxicity on the N assimilation of transplanted seedlings of both varieties, as their leaves and roots presented reduced GS activities and SP contents, and increased GDH activities, and NH₄⁺ and FAA content. Glufosinate phytotoxicity on the N assimilation of transplanted seedlings reached a plateau at 11 to 20 d. further, GDH in roots and GS in leaves were still significantly different at 70 d. Meanwhile, ZS11 might be more sensitive to glufosinate than D148 since ZS11 had more variation than D148 at the same treatment, and the overdose of glufosinate more strongly inhibited N assimilation than the recommended dose. Therefore, it is essential to apply a suitable glufosinate dose to the transplanted variety, to minimize adverse effects on crops and the environment.

Abbreviations: N, Nitrogen; GS, Glutamine synthetase; GDH, glutamate dehydrogenase; NH₄⁺, ammonium; FAA, free amino acids; SP, soluble proteins; DW, Dry weight; ANOVA, one-way analysis of variance; NO₃^?, nitrate; OECD, Organisation for Economic Co-operation and Development; PPT, phosphinothricin; USEPA, United States Environmental Protection Agency     

Changes in growth and nitrogen assimilation in barley seedlings under cadmium stress

Barley (Hordeum vulgare L. cv. Martin) plants grown in solution culture, were exposed to increasing cadmium (Cd) concentration (0, 5, 10, 25, 50, and 100 μM) for a duration of 12 days. The sequence of important biochemical steps of nitrate (NO₃) assimilation were studied in roots and shoots as a function of external Cd concentration. Cadmium uptake in roots and shoots increased gradually with Cd concentration in the medium. This Cd accumulation lowered substantially root and shoot biomass. The nitrate reductase (NR, EC 1.6.6.1) and nitrite reductase (NiR, EC 1.6.6.4) activities declined under Cd stress. Concurrently, tissue NO₃ contents and xylem sap NO₃ concentration were also decreased in Cd‐treated plants. These results suggest that Cd could exert an inhibitory effect on the assimilatory NO₃ reducing system (NR and NiR) through a restriction of NO₃ availability in the tissues. We therefore examined, in short‐term experiments (12 h), the impact of Cd on NO₃ uptake and the two reductases in nitrogen (N)‐starved plants that were pretreated or not with Cd. It was found that Cd induced inhibition of both NO₃ uptake and activities of NR and NiR, during NO₃ induction period. The possible mechanisms of Cd action on NO₃ uptake are proposed. Further, in Cd‐grown plants, the glutamine synthetase (GS, EC 6.3.1.2) showed a decreasing activity both in shoots and roots. However, increasing external Cd concentration resulted in a marked enhancement of glutamate dehydrogenase (NADH‐GDH, EC 1.4.1.2) activity, coupled with elevated levels of ammonium (NH₄ in tissues. On the other hand, the total protein content in Cd‐treated plants declined with a progressive and substantial increase of protease activity in the tissues. These findings indicate that under Cd stress the usual pathway of NH₄ assimilation (glutamine synthetase/glutamate synthase) can switch to an alternative one (glutamate dehydrogenase). The changes in all parameters investigated were concentration‐dependent and more marked in roots than shoots. The regulation of N absorption and assimilation by Cd in relation to growth and adaptation to stress conditions are discussed.     

Effects of N‐source,light intensity and temperature on nitrogen metabolism of bahiagrass

Under greenhouse conditions, a study was made on the effects of nitrogen (N) source (N)O₃ or NH₄), mode of application (single vs. split) and nitrification inhibition on the N‐uptake and metabolism, of bahiagrass.

Variations in light and temperature in the greenhouse affected the N‐metabolism of bahiagrass plants. Nitrate fed plants had nitrate reductase activity (NRA) pattern different from that of NH₄‐fed plants. Amino‐N accumulation patterns were similar for plants under both N‐sources, although amino‐N levels in leaves of NH₄‐fed plants were much smaller than that of NO₃ plants. Nitrate accumulation in leaves showed inverse trend to that of roots in plants fed both NO₃ or NH₄. To the sharp peaks in NO₃ levels in roots due to increases in light and temperature corresponds a sharp decrease of its levels in leaves.

For both both NO₃ or NH₄ treatments, soluble‐N accumulated most in the rhizomes of bahiagrass plants, whereas protein N accumulated most in leaves, suggesting that rhizomes had a buffering effect on the NO₃ fluxes to leaves. This presumably resulted in a lag in the NRA response of the NO₃‐fed plants to increases in light and temperature.     

Ionic Activity in Soil Solution as Affected by Application Of Newsprint and Nitrogen Sources

The influence of noncomposted ground newsprint (GNP) and nitrogen (N) source on corn (Zea Mays L.) dry matter production, grain yield, and soil chemical properties has been previously reported (Lu et al. 1995). The effects of GNP and N source on soil solution ionic activities at 40 days after planting in a field study; seed germination and extractable aluminum (Al) in GNP in laboratory studies were evaluated to determine their effects on corn seedling stunting and nutrient imbalances during early growth stages. Ammonium nitrate (NH₄NO₃), urea, anhydrous ammonia (NH₃), or poultry litter (PL) were the N sources used in the field study to adjust the C:N ratio of the GNP to ≤ 30:1. In laboratory experiments, cotton, soybean, and corn germination at seven days was not affected by N source or GNP applied at a rate of 2.44 kg C/m² soil; but N source did influence the dry weight of corn root/shoot ratio at 21 days. The Al extracted from GNP increased as the NH₄OH concentration in the extracting solution increased and followed a quadratic relationship with an r² of 0.90. The σα_ca/σα_cation ratio in soil solution for all N sources was greater than the 0.15 reported by Bennett and Adams (1970a), where incipient NH₃ phytotoxicity can occur. At 40 days after planting, a two-fold increase in soil monomeric Al (σAlα_mon.) ionic activity and a five-fold increase in soil P (σPα) ionic activity were measured in GNP treatments as compared to no N GNP treatments. When NH₃ was the N source used to adjust the C:N ratio of GNP, the σAlα_mon. ionic activities were increased by a factor of five as compared to NH₃ applied alone. When PL was the N source, the Al_T (σAlα_species) ionic activity was 119 mmol L^?1 compared to the σAlα_mon. ionic activity of 0.53 mmol L^?1. It appears that σAlα_mon. ionic activity induced nutrient disorder and caused severe stunting of corn seedlings during early season growth. The relatively high water-soluble organic carbon in PL (18 percent) may have acted as a chelating agent to reduce the σAlα_mon. ionic activities in the GNP, or as a soluble carbon source for increasing microbial utilization of all the N, thus slowing the formation and accumulation of phytotoxic levels of by-products.     

Nutrient solution nitrogen form and barley yellow dwarf virus disease tolerance in oat and wheat

Abstract

The form of nutrient solution nitrogen (either NH₄‐N or NO₃‐N or mixtures of the two) provided to plants influences the severity of many crop diseases. This greenhouse study was conducted to determine how growth, grain yield, and yield components of oat (Avena sativa L.) and wheat (Triticum aestivum L.) plants given nutrient solutions containing different ratios of NO₃‐N to NH₄‐N would react to barley yellow dwarf virus (BYDV) infection. Fifteen‐day‐old seedlings (2nd leaf stage) were either infected with BYDV (PAV strain) or left uninfected. Nutrient solution treatments (started 19 d after germination) provided three ratios of NO₃‐N to NH₄‐N (100% NO₃, 50:50 NH₄:NO₃, or 100% NH₄) for a 30‐d period, after which plant height and tillers plant^?1 were measured. Oat and wheat plants given NH₄ had fewer tillers than plants given the other nutrient solution treatments. BYDV‐infected oat and wheat plants were shorter than uninfected plants. All pots then received NO₃ nutrient solution until plant maturity, after which days to anthesis, primary tiller height, grain yield and yield components were measured. In the NH₄ nutrient solution treatments, BYDV infection significantly reduced individual kernel weight in oat and primary tiller height in wheat. These same measures were not significantly affected by BYDV infection in the NO₃ or NH₄NO₃ nutrient solution treatments. There were no other significant nutrient solution by BYDV infection interactions for any other dependent variable measured. Nutrient solution treatments had no significant effect on grain yield, but BYDV infection reduced grain yield by 45% in oat and 46% in wheat. In conclusion, nutrient solution N form interacted with BYDV infection to alter disease tolerance in oat (kernel weight) and wheat (primary tiller height), but these alterations had no effect in ameliorating grain yield loss caused by BYDV disease.     

Growth and chloroplast ultrastructure of two citrus rootstock seedlings in response to ammonium and nitrate nutrition 1

Abstract

It is important to understand the differential response of citrus rootstock to various rates of nitrogen (N) forms in order to evaluate the tree response to N availability under different production conditions. In this study, the effects of N sources (NH₄‐N or NO₃‐N), and rates (5, 15, 45, and 135 mg N L^?1) on two citrus rootstock seedlings (Swingle citrumelo, SC; Cleopatra mandarin, CM) growth (110 d) and N concentrations in various parts of seedling were investigated in a nutrient solution experiment. The effects of N nutrition on the chloroplast ultrastructure of leaves were examined at the end of the experiment. Rootstock and N sources significantly influenced the growth of leaves, stems, and roots. The growth of all seedling parts of both rootstocks decreased with an increase in concentration of NH₄‐N in the solution. In contrast, there was a positive relationship between the seedling growth and the concentration of NO₃‐N in the nutrient solution with marked response observed as the NO₃‐N concentration increased from 5 to 15 mg L^?1 The experiment demonstrated a distinct growth suppression effect with an increase in concentration of NH₄‐N in the nutrient solution, particularly SC rootstock. Chloroplast ultra‐structure of the leaves showed evidence of injury of the seedlings which received N entirely as NH₄ ⁺ form, but the injury was not seen when the seedlings received N as the NO₃‐N form. The disruption of chloroplast ultrastructure increased with increased rate of NH₄‐N. The most conspicuous characteristic of ammonium toxicity was the massive accumulation of strands granules and phytoferritin which is clearly an evidence of NH₃ toxicity. The results are important for understanding the implications of N source on seedling growth and chloroplast structure of citrus leaves.     

Effect of nitrogen source on aluminum toxicity in nonmycorrhizal and ectomycorrhizal pitch pine seedling

The effect of elevated nitrate [(NO₃‐nitrogen (N)] or ammonium (NH₄)‐N on the response of nonmycorrhizal (NM) and ectomycorrhizal (ECM) pitch pine (Pintis rigida Mill.) seedlings to aluminum (Al) was determined in experiments in which N was increased three times above ambient levels. Seedlings with and without the mycorrhizal fungus Pisolithus tinctorius (Pers.) Coker & Couch were grown in sand irrigated with nutrient solution (pH 3.8) containing 0, 10, or 20 mg Al L^‐1 (0, 370, or 740 μM Al). The nutrient solution simulated that for the sandy, nutrient‐poor soil of the New Jersey Pine Barrens. Elevated NO₃‐N had no significant effect on Al toxicity in NM seedlings, but Al toxicity at ambient NH₄‐N was ameliorated by elevated NH₄‐N. Symptoms of Al toxicity in roots (thick and stunted) of ECM seedlings at ambient N levels were reduced by elevated NH₄‐N and absent at elevated NO₃‐N. When N was elevated by an increase in NO₃‐N or NH₄‐N, uptake of N and relative increases in total biomass were greater in ECM than in NM seedlings.     

Response of wheat and maize to different nitrogen sources: II. Nitrate reductase and glutamine synthetase enzyme activities,and plastid pigment content

Glutamine synthetase and nitrate reductase enzyme activities occurred both in roots and leaves of maize (Zea mays L., hybrid Pioneer 3737) and wheat (Triticum aestivum L., cultivar Jantar) plants grown on different nitrogen (N) sources. Enzyme activities and plastid pigment content in maize plants were higher in the treatments with a mixture of nitrate (NO₃) and ammonium (NH₄) than with either N source alone. In wheat plants, plastid pigment content, nitrate reductase activity, and root glutamine snynthetase activity were higher in the treatments where NO₃ alone was applied to the nutrient medium.     

Determining potential glutamine synthetase and glutamate dehydrogenase activity in soil

Ammonium (NH₄⁺), an important nitrogen (N) source for microorganisms, is assimilated via two major pathways. One route is catalyzed by glutamate dehydrogenase (GDH), while the other mechanism involves two enzymes, glutamine synthetase (GS) and glutamate synthase (GOGAT). The GS/GOGAT enzyme system requires more energy to operate, but has a much higher affinity for NH₄⁺ than GDH. We describe procedures to determine potential GS and GDH activity in soil samples. GS and GDH are intracellular enzymes. We used chloroform fumigation to make cell membranes permeable for substrates and products of the enzymes. Fumigation for 4 h increased GS activity almost ten-fold compared to the unfumigated control. Under optimized assay conditions, GS activity increased linearly for at least 80 min, indicating that the substrates were not limiting. In contrast to what was found for GS activity, direct addition of substrates to the soil to assay GDH activity did not result in a linear increase in GDH activity over time. A linear response for 3 h, however, resulted when the soil samples were first extracted with buffer solution and the reagents were added after centrifugation. The differences between the assays explain why fumigation for 3 d prior to the assay increased GDH activity by only 60%. In a microcosm study with glucose and NH₄⁺ addition, the activity of the two enzymes depended on the carbon (C) to N ratio of the amendment. With increasing C to N ratios from 5 to 120, GS activity doubled, while C to N ratios higher than 120 did not further increase GS activity. In contrast, GDH activity decreased by 13% with increasing C to N ratios from 5 to 200. The GDH to GS activity ratio in soil may therefore yield valuable information about the availability of N relative to C at a specific time.     

EFFECTS OF CALCIUM ON LIGNIFICATION RELATED PARAMETERS IN SODIUM CHLORIDE-STRESSED SOYBEAN ROOTS

The effects of exogenous calcium (Ca²⁺) on root growth and lignification-related parameters – phenylalanine ammonia-lyase (PAL) and peroxidases (POD) activities, hydrogen peroxide (H₂O₂) and lignin contents – in roots of NaCl-stressed soybean seedlings were analyzed. Three-day-old seedlings were cultivated in half-strength Hoagland's solution (pH 6.0) with or without 5 mM calcium nitrate [Ca(NO₃)₂] and 50 to 200 mM sodium chloride (NaCl) in a growth chamber (25°C, 12/12 h light/dark photoperiod, irradiance of 280 μmol m^?2 s^?1) for 24 h. In general, results showed that the absence of Ca²⁺ reduced root growth and increased lignification of soybean seedlings grown in NaCl-free nutrient solution. NaCl reduced the root growth and all lignification-related parameters. Root growth, PAL and POD activities and hydrogen peroxide (H₂O₂) contents were more affected after NaCl treatments without Ca²⁺ in the nutrient solution. At 5 mM, Ca²⁺ did not alleviate the deleterious effects of NaCl on lignification-related parameters.     

Tomato growth and nutrient uptake patterns as influenced by nitrogen form and light intensity 1

Tomato plants were grown in sand culture with NH₄ or NO₃ forms of N and at two levels of light. Plants were harvested at 0, 5, 9, or 12 days after starting treatments. NH₄‐N nutrition reduced growth, suppressed K, Ca, and Mg accumulation in shoot, increased P and N content and markedly reduced K, Ca, and Mg uptake per unit of root surface. Reduced light level decreased the toxic effects of NH₄ and markedly decreased NH₄ accumulation in shoots.     

Ammonium assimilation in rice based on the occurrence of 15N and inhibition of glutamine synthetase activity

Assimilation of ammonium (NH₄) into free amino acids and total reduced nitrogen (N) was monitored in both roots and shoots of two‐week old rice seedlings supplied with 5 mM 99% (¹⁵NH₄)₂SO₄ in aerated hydroponic culture with or without a 2 h preincubation with 1 mM methionine sulfoximine (MSX), an inhibitor of glutamine synthetase (GS) activity. ¹⁵NH₄ was not assimilated into amino acids when the GS/GOGAT (glutamate synthase) cycle was inhibited by MSX. Inhibition of glutamine synthetase (GS) activity in roots with MSX increased both the amount of NH₄ and the abundance of ¹⁵N labeled NH₄. In contrast, the amount of Gln and Glu, and their proportions as ¹⁵N, decreased in roots when GS activity was inhibited. This research confirms the importance of GS/GOGAT in NH₄ assimilation in rice roots.

¹⁵N‐labeled studies indicate that NH₄ ions incorporated by roots of rice are transformed primarily into glutamine (Gln) and glutamic acid (Glu) before being converted to other amino acids through transamination (15). The formation of amino acids such as aspartic acid (Asp) and alanine (Ala) directly from free NH₄ in roots also has been reported (4,15). Translocation of free NH₄ to plant shoots, based on the concentration of free NH₄ in xylem exudate, has been reported in tomato (13), although NH₄ in shoots primarily originates from nitrate reduction in the shoot. Photorespiration also can contribute to the accumulation of NH₄ in leaves (7).

The GS/GOGAT cycle appears to be primarily responsible for the assimilation of exogenously supplied NH₄ and NH₄ derived from nitrate reduction in leaves, as well as NH₄ derived from photorespiration (2,3,6,8). Genetic evidence cited to support this conclusion includes the lethal effect of photorespiratory conditions on plant mutants deficient in chloroplast‐localized GS and GOGAT activities (2,3,9), and the rapid accumulation of free NH₄ in GS‐deficient mutants under photorespiratory conditions (2,3,5).

The present study was initiated to quantify the in vivo amino acid synthesis in rice roots and shoots by analysis of ¹⁵N labeling, and should provide a more complete understanding of this important system for NH₄ utilization.     

利用gs1.1和gs1.2突变体研究外源蔗糖对高铵胁迫拟南芥碳氮代谢的影响

以拟南芥野生型Col-0、谷氨酰胺合成酶敲除突变体gs1.1和gs1.2为实验材料,采取土培试验,比较正常培养液(4 mmol/L NH4⁺)培养(CK)、正常培养液(4 mmol/L NH4⁺)下外源添加5%蔗糖(T1)、高NH4⁺胁迫(20 mmol/L)(T2)以及高NH4⁺胁迫(20 mmol/L)下外源添加5%蔗糖(T3)对拟南芥各株系各生理指标的影响;通过测定地上部分的鲜重、叶绿素、游离NH4⁺、可溶性糖、可溶性蛋白、谷氨酰胺合成酶(GS)、谷氨酸脱氢酶(GDH)、矿质元素含量等指标,研究外源蔗糖对NH4⁺胁迫拟南芥碳氮代谢的影响。结果表明,高NH4⁺胁迫下,拟南芥生长受到严重的抑制,鲜重、GS、GDH酶活性降低,游离NH4⁺含量、叶绿素含量、可溶性糖和可溶性蛋白含量增加,植株的N、P、K、Ca的含量增加,Mg、Fe的含量减少,其中gs1.1和gs1.2在高NH4⁺处理下受到的抑制比Col-0更为显著。外源添加5%蔗糖显著缓解了高NH4⁺毒害,提高了可溶性糖和可溶性蛋白含量,提高了GS和GDH的活性,降低了叶绿素和游离NH4⁺的含量,提高了植株体内的N、P、K、Ca,Mg的含量,降低了植株Fe的含量,其中,外源蔗糖对gs1.1和gs1.2高NH4⁺毒害的缓解更为显著。