Response of Ammonia Assimilation in Cucumber Seedlings to Nitrate Stress

The influence of increased nitrate concentration—14 (control) and 140 mmol L⁻¹ (T)—in hydroponic culture on ammonia assimilation in cucumber (Cucumis sativus L. cv. Xintaimici) seedlings was investigated. The results showed that NH₃ accumulation in the roots and leaves of T seedlings increased significantly, indicating that NH₃ toxicity might be involved in nitrate stress. Under control conditions, GS and GOGAT activity were much higher in the leaves than in the roots, whereas GDH activity was much higher in the roots than in the leaves. Correlation analysis showed that NH₃ concentration had a strong negative linear relationship with GDH activity in the roots but had a strong negative linear relationship with GS and GOGAT activity in the leaves. These results indicate that NH₃ might be assimilated primarily via GDH reaction in the roots and via GS/GOGAT cycle in the leaves. Short-term nitrate stress resulted in the increase of GS and GOGAT activity in the roots and GDH activity in the leaves of T seedlings, indicating possible shifts in ammonia assimilation from the normal GDH pathway to GS/GOGAT pathway in the roots and from the normal GS/GOGAT pathway to the GDH pathway in the leaves under nitrate stress, but with the increase of treatment time, GS, GOGAT, and GDH activity in the roots and leaves of T seedlings decreased possibly due to low water potential and NH₃ toxicity.     

The effects of grafting on glycolysis and the tricarboxylic acid cycle in Ca(NO<Subscript>3</Subscript>)<Subscript>2</Subscript>-stressed cucumber seedlings with pumpkin as rootstock

In this research, we investigated the effects of grafting on intermediate metabolites and key enzymes of glycolysis and the tricarboxylic acid (TCA) cycle in self-grafted and salt-tolerant pumpkin rootstock-grafted cucumber seedlings supplied with nutrient solution and subjected to 80 mM Ca(NO₃)₂ stress for 6 days. Ca(NO₃)₂ stress induced accumulation of 3-phosphoglycerate (3-PGA) and phosphoenolpyruvate (PEP) in the leaves of self-grafted cucumber seedlings and enhanced the activities of phosphoenolpyruvate carboxylase (PEPC) and enolase (ENO). Succinic acid and malic acid contents and isocitrate dehydrogenase, succinate dehydrogenase (SDH), and malate dehydrogenase (MDH) activities in self-grafted seedlings were significantly decreased by Ca(NO₃)₂ stress. In addition, activities of PEPC, ENO, SDH, and MDH and contents of glycolysis intermediate metabolites (citric, succinic, and malic acids) were significantly higher in leaves of rootstock-grafted seedlings compared with those in self-grafted seedlings under saline conditions. Furthermore, leaf adenosine triphosphate (ATP) content of rootstock-grafted seedlings was relatively higher than that in self-grafted plants under salt stress, with an opposite effect observed on adenosine diphosphate content. These results indicate that rootstock grafting alleviates Ca(NO₃)₂ stress-induced inhibition of the glycolytic pathway and the TCA cycle in cucumber seedling leaves, which may aid the respiratory metabolism of cucumber seedlings and help maintain a high ATP synthesis level, thereby increasing the biomass of cucumber seedlings and enhancing their salt tolerance.     

Nitrogen metabolism in leaves of a tank epiphytic bromeliad: characterization of a spatial and functional division

The leaf is considered the most important vegetative organ of tank epiphytic bromeliads due to its ability to absorb and assimilate nutrients. However, little is known about the physiological characteristics of nutrient uptake and assimilation. In order to better understand the mechanisms utilized by some tank epiphytic bromeliads to optimize the nitrogen acquisition and assimilation, a study was proposed to verify the existence of a differential capacity to assimilate nitrogen in different leaf portions. The experiments were conducted using young plants of Vriesea gigantea. A nutrient solution containing NO₃⁻/NH₄⁺ or urea as the sole nitrogen source was supplied to the tank of these plants and the activities of urease, nitrate reductase (NR), glutamine synthetase (GS) and glutamate dehydrogenase (NADH-GDH) were quantified in apical and basal leaf portions after 1, 3, 6, 9, 12, 24 and 48 h. The endogenous ammonium and urea contents were also analyzed. Independent of the nitrogen sources utilized, NR and urease activities were higher in the basal portions of leaves in all the period analyzed. On the contrary, GS and GDH activities were higher in apical part. It was also observed that the endogenous ammonium and urea had the highest contents detected in the basal region. These results suggest that the basal portion was preferentially involved in nitrate reduction and urea hydrolysis, while the apical region could be the main area responsible for ammonium assimilation through the action of GS and GDH activities. Moreover, it was possible to infer that ammonium may be transported from the base, to the apex of the leaves. In conclusion, it was suggested that a spatial and functional division in nitrogen absorption and NH₄⁺ assimilation between basal and apical leaf areas exists, ensuring that the majority of nitrogen available inside the tank is quickly used by bromeliad's leaves.     

Effects of 5-aminolevulinic acid on nitrogen metabolism and ion distribution of watermelon seedlings under salt stress

The effects of foliar spray application of 5-aminolevulinic acid (ALA) on the growth, nitrogen metabolism, and ion distribution of salt-stressed watermelon (Citrullus lanatus (Thunb.) Matsum. and Nakai) seedlings were investigated. Supplementation of the nutrient solution with 100 mM NaCl significantly reduced leaf and root biomass of watermelon plants. Foliar application of 1.25 mM ALA significantly alleviated the inhibition of plant growth under salt stress. Salinity induced significant accumulation of nitrate, ammonium, and soluble protein and a significant decrease in the activities of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT), and glutamate dehydrogenase (GDH) in watermelon plants. However, ALA significantly increased the activities of NR, GS, GOGAT, and GDH, but decreased the ammonium content and NiR activity. In addition, salt stress resulted in significant accumulation of Na⁺ and Cl^? in plants, but decreased the contents of K⁺ and Mg²⁺. Application of ALA alleviated the salt stress-induced ion toxicity, and increased the contents of K⁺ and Mg²⁺. ALA also increased soluble protein and proline contents in salt-stressed watermelon plants. These results indicated that application of ALA alleviated the accumulation of Na⁺ and Cl^? in salt-stressed watermelon plants, especially through regulating nitrogen metabolism and ion distribution, which were associated with an improvement in plant growth.     

外源褪黑素对高温胁迫条件下黄瓜幼苗氮代谢的影响

以‘津优4号’（热敏感型）和‘美国保尔’（耐热型）黄瓜幼苗为试材,研究了叶面喷施褪黑素对高温胁迫条件下黄瓜幼苗氮代谢的影响。结果显示,高温胁迫下,（1）两种黄瓜幼苗硝态氮含量先升高后降低,‘津优4号’总氮和氨态氮含量先下降后持续升高,而‘美国保尔’总氮和氨态氮含量持续上升;（2）两种黄瓜幼苗硝酸还原酶（NR）活性均先上升后下降,而谷氨酰胺合成酶（Gs）、谷氨酸合成酶（GOGAT）和谷氨酸脱氢酶（GDH）均持续下降,‘美国保尔’的4种酶活性下降幅度显著低于‘津优4号’。研究结果表明,叶面喷施褪黑素可有效缓解高温胁迫对NR、GS、GOGAT和GDH的抑制作用,显著增加硝态氮含量,降低氨态氮含量,减轻氨态氮积累对黄瓜幼苗造成的毒害作用,增强高温胁迫条件下黄瓜幼苗氮素的代谢能力,减轻高温胁迫对黄瓜幼苗造成的伤害,提高黄瓜幼苗抵御高温胁迫的能力。     

纯化腐植酸对氮胁迫下黄瓜幼苗生长和氮代谢的影响

为研究纯化腐植酸(PHA)在不同水平氮胁迫下对黄瓜植株生长和氮代谢的影响,探明PHA对逆境胁迫的缓解作用机制,采用水培方式,选用新泰密刺为供试品种,以正常氮水平(11 mmol·L^-1 NO₃^-)为对照,进行低氮(1.0 mmol·L^-1 NO₃^-)和高氮(101 mmol·L^-1 NO₃^-)胁迫处理,研究纯化腐植酸对氮胁迫下黄瓜幼苗生长和氮代谢的影响.结果表明: 低氮和高氮胁迫均抑制了黄瓜幼苗生长,株高、茎粗、叶面积、干物质积累量均低于正常氮水平处理.施用纯化腐植酸促进了正常氮水平及低氮胁迫下黄瓜干物质积累,在高氮胁迫下差异不显著.PHA影响了黄瓜幼苗NO₃^-的吸收,呈低氮下促进、高氮下抑制吸收的趋势;PHA显著降低了低氮、高氮胁迫下根系和叶片中的铵态氮含量;与正常氮水平相比,低氮、高氮胁迫下根系和叶片的硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、谷氨酸脱氢酶(GDH)活性及根系中亚硝酸还原酶(NiR)活性均显著降低,PHA不同程度地提高了NR、NiR、GS、GOGAT、GDH活性,还提高了根系和叶片中游离氨基酸、可溶性蛋白的含量.综上,添加PHA缓解了氮胁迫对黄瓜幼苗生长的抑制作用.     

NaCl对水稻谷氨酸合酶和谷氨酸脱氢酶的胁迫作用

在ＮａＣｌ的胁迫下,水稻幼苗根和叶的谷氨酸合酶和谷氨酸脱氢酶的活性随着营养液中的ＮａＣｌ浓度的升高而降低;游离ＮＨ４＾＋在叶中积累,在根中未见明显变化。与根相比,叶对ＮａＣｌ的胁迫作用更为敏感。叶的ＮＡＤＨ－ＧＯＧＡＴ和ＮＡＤＨ－ＧＤＨ活性在ＮａＣｌ胁迫降低的程度明显大于根。无论是否有ＮａＣｌ存在,根的ＮＡＤＨ－ＧＤＨ活性明显高于叶。ＧＳ／ＧＤＨ比值分析提示,对对照下,根中的ＮＨ４＾存在,根的ＮＡ     

Effect of applied nitrate on enzymes of ammonia assimilation in nodules ofCicer arietinum L.

Summary The relationship between N₂-fixation, nitrate reductase and various enzymes of ammonia assimilation was studied in the nodules and leaves ofC. arietinum. In the nodules of the plants growing on atmospheric nitrogen, maximum activities of glutamine synthetase (GS), glutamate synthase (GOGAT), glutamate dehydrogenase (GDH), asparagine synthetase (AS) and aspartate aminotransferase (AAT) were recorded just prior to maximum activity of nitrogenase. In nitrate fed plants, the first major peak of GDH and AS coincided with that of nitrate reductase in the nodules. With the exception of AS, application of nitrate decreased the activities of all these enzymes in nodules but not in leaves. Activities of GS, GOGAT and AAT were affected to much greater extent than that of GDH. On comparing the plants grown without nitrate and those with nitrate, the ratios of the activities of GDH/GS and GDH/GOGAT in nitrate given plants, increased by 4 and 12 fold, respectively. The results presented in this paper suggest that in nodules of nitrate fed plants, assimilation of ammonia via GDH assumes much greater importance.     

Biochar improved nodulation and nitrogen metabolism of soybean under salt stress

To investigate salt stress and biochar application effects on nodulation and nitrogen metabolism of soybeans (Glycine max cv. M7), an experiment was conducted under the control condition. The treatments comprised three biochar rates (non, 50 and 100 g kg^?1 soil) and three salinities (0, 5 and 10 dS m^?1 NaCl), with four replications of treatments. Salt stress diminished the number of nodules and their weights in the soybean roots. Nitrogen content and metabolism decreased in nodules, roots and shoots, while reducing the activity of glutamate dehydrogenase (GDH), glutamine synthetase (GS), glutamine oxoglutarate aminotransferase (GOGAT) and nitrate reductase (NR). Also, salinity brought down root and shoot weight, total plant biomass, chlorophyll content, leaf area (LA) and rubisco activity in the soybean. On the other hand, application of biochar improved nodulation, nitrogen content, rubisco activity, GDH, GS, GOGAT and NR activities in different parts of the soybean and nodules under salt stress, and consequently improved chlorophyll content, LA, root and shoot weight. Both the 50 and 100 g kg^?1 biochar rates showed similar effects in improving nitrogen metabolism and plant performance under salt stress. Generally, biochar increased nodulation and nitrogen metabolism of the soybean under saline conditions.     

Grafting onto Cucurbita moschata rootstock alleviates salt stress in cucumber plants by delaying photoinhibition

To determine how the use of a given rootstock can influence the functioning of the photosynthetic apparatus of the scion under salt stress, the growth, gas exchange, photosystem II (PSII) efficiency, xanthophyll cycle, and chloroplast ultrastructure of nongrafted, self-grafted, and pumpkin-grafted (hereafter referred to as rootstock-grafted) cucumber (Cucumis sativus L.) plants were investigated at day 15 after being treated with 90 mM NaCl. The reductions in plant growth of the rootstock-grafted plants were lower than those of the nongrafted and self-grafted plants under 90 mM NaCl. The net photosynthetic rate, stomatal conductance, maximal and effective quantum yield of PSII photochemistry, photochemical quenching coefficient, and effective quantum-use efficiency of PSII in the light-adapted state of the nongrafted and self-grafted plants were significantly decreased under 90 mM NaCl. However, these reductions were alleviated when the cucumber plants were grafted onto the pumpkin (Cucurbita moschata Duch.) rootstock. The intercellular CO₂ concentrations were significantly increased in the nongrafted and self-grafted plants under 90 mM NaCl, whereas it was decreased in the rootstock-grafted plants. Nonphotochemical quenching (NPQ) and the deepoxidation state of the xanthophyll cycle were significantly increased under 90 mM NaCl, particularly in the rootstockgrafted plants, suggesting the rootstock-grafted plants had higher potential to dissipate excess excitation energy and reduce the probability of photodamage to PSII. Under 90 mM NaCl, the number of grana was reduced, the thylakoids were swollen, and starch granules accumulated in all plants. However, the damage of chloroplast ultrastructure was alleviated in the rootstock-grafted plants. Taken together, the use of C. moschata rootstock alleviated salt stress in cucumber plants by delaying photoinhibition, probably due to a lower incidence of both stomatal and nonstomatal factors limiting photosynthesis.     

Ammonium formation and assimilation in P(SARK)∷IPT tobacco transgenic plants under low N

Wild Type (WT) and transgenic tobacco plants expressing isopentenyltransferase (IPT), a gene encoding the enzyme regulating the rate-limiting step in cytokinins (CKs) synthesis, were grown under limited nitrogen (N) conditions. We analyzed nitrogen forms, nitrogen metabolism related-enzymes, amino acids and photorespiration related-enzymes in WT and P_SARK∷IPT tobacco plants. Our results indicate that the WT plants subjected to N deficiency displayed reduced nitrate (NO₃⁻) assimilation. However, an increase in the production of ammonium (NH₄⁺), by the degradation of proteins and photorespiration led to an increase in the glutamine synthetase/glutamate synthase (GS/GOGAT) cycle in WT plants. In these plants, the amounts of amino acids decreased with N deficiency, although the relative amounts of glutamate and glutamine increased with N deficiency. Although the transgenic plants expressing P_SARK∷IPT and growing under suboptimal N conditions displayed a significant decline in the N forms in the leaf, they maintained the GS/GOGAT cycle at control levels. Our results suggest that, under N deficiency, CKs prevented the generation and assimilation of NH₄⁺ by increasing such processes as photorespiration, protein degradation, the GS/GOGAT cycle, and the formation of glutamine.     

Heme oxygenase up-regulation under salt stress protects nitrogen metabolism in nodules of soybean plants

The behaviour of enzymes involved in nitrogen metabolism, as well as oxidative stress generation and heme oxygenase gene and protein expression and activity, were analysed in soybean (Glycine max L.) nodules exposed to 50, 100 and 200 mM NaCl concentrations. A significant increase in lipid peroxidation was found with 100 and 200 mM salt treatments. Moreover, superoxide dismutase, catalase and peroxidase activities were decreased under 100 and 200 mM salt. Nitrogenase activity and leghemeoglobin content were diminished and ammonium content increased only under 200 mM NaCl. At 100 mM NaCl, glutamine synthetase (GS) and NADH-glutamate dehydrogenase (GDH) activities were similar to controls, whereas a significant increase (64%) in NADH-glutamate synthase (GOGAT) activity was observed. GS activity did not change at 200 mM salt treatment, but GOGAT and GDH significantly decreased (40 and 50%, respectively). When gene and protein expression of GS and GOGAT were analysed, it was found that they were positively correlated with enzyme activities. In addition, heme oxygenase (HO) activity, protein synthesis and gene expression were significantly increased under 100 mM salt treatment. Our data demonstrated that the up-regulation of HO, as part of antioxidant defence system, could be protecting the soybean nodule nitrogen fixation and assimilation under saline stress conditions.     

Studies on ammonium-assimilating enzymes of Platymonas striata Butcher (Prasinophyceae)

Platymonas striata Butcher displays significant levels of glutamate synthase (GS) (EC 2.6.1.53) and glutamine synthetase (GOGAT) (EC 6.3.1.2.), but very low levels of glutamate dehydrogenase (GDH) (EC 1.4.1.4). This suggests that the GS/GOGAT pathway is important for nitrogen assimilation. The in vitro rates of enzyme activity can however only account for about 10% of the in vivo rates of nitrogen assimilation. Nitrogen-starvation reduced GS activity to undetectable levels. On nitrate or ammonium ion refeeding the cellular GS activity was rapidly restored, and reached levels of 56% and 91% greater than the unstarved values 24h after refeeding nitrate or ammonium respectively.Abbreviations NAR nitrate reductase - NIR nitrate reductase     

叶施NO对NaCl胁迫下红砂幼苗叶片和根系中氮代谢酶及营养物质的影响

以当年生红砂(Reaumuria soongorica)幼苗为材料,采用盆栽实验,考察叶面喷施不同浓度(0、0.01、0.10、0.25、0.50、1.00 mmol·L^-1)NO供体硝普钠 (SNP) 对NaCl(300 mmol·L^-1)胁迫下红砂根、叶中可溶性蛋白、游离氨基酸和硝态氮含量,以及谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、硝酸还原酶(NR)活性的影响,并采用主成分分析和隶属函数法筛选NO对NaCl胁迫缓解效应的氮代谢指标和最佳NO浓度,以探讨外源NO对NaCl 胁迫下红砂缓解效应的氮代谢响应机制。结果表明：(1)在300 mmol·L^-1 NaCl胁迫处理下,红砂幼苗根、叶中可溶性蛋白、硝态氮含量以及GS、GOGAT、NR活性均比对照显著下降。(2)外源NO能显著提高盐胁迫下红砂叶、根中GS、GOGAT、NR活性和硝态氮含量,增加根中可溶性蛋白和游离氨基酸含量。(3)NR和GOGAT活性可用于评价NO对NaCl胁迫下红砂幼苗的缓解作用,外源NO(SNP)对红砂幼苗在NaCl胁迫下的缓解效果强弱表现为0.25 mmol·L^-1> 0.50 mmol·L^-1> 0.10 mmol·L^-1> 1.00 mmol·L^-1> 0.01 mmol·L^-1。研究发现,300 mmol·L^-1 NaCl胁迫显著抑制了红砂幼苗氮代谢,外源NO(SNP)有助于提高盐胁迫下红砂NR活性,加快硝态氮转化为铵态氮,促进红砂叶片和根中GS/GOGAT对转化物的同化,从而增强红砂幼苗的耐盐性,并以0.25 mmol·L^-1SNP处理时缓解作用最佳;NR和GOGAT活性可作为NO缓解盐胁迫的评价指标。     

Effect of ammonium salt on enzymes of ammonium assimilation in maize seedlings

Glutamate dehydrogenase (GDH E.C. 1.4.1.2.4), glutamine synthetase (GS E.C. 6.3.1.2) and glutamate synthase (glutamine oxoglutarate amino transferase, GOGAT E.C. 2.6.1.53) activities, protein and organic nitrogen contents and growth of roots and shoots of maize seedlings raised in dark at 25±2°C in half strength Hoagland’s solution containing different ammonium salts as source of nitrogen, were determined to assess the contribution of alternate pathways in ammonium assimilation. Ammonium nitrate or in some cases ammonium chloride appeared to be the best source for both root and shoot growth and for increase in protein, total nitrogen and the enzymes of ammonium assimilation. In roots, NH₄-nitrogen appeared to be assimilated by both GDH as well as GS-GOGAT pathways specially in the dark grown seedlings, while in shoots it was primarily by GS-GOGAT pathway.     

Kinetic flux profiling dissects nitrogen utilization pathways in the oleaginous green alga Chlorella protothecoides

As a promising candidate for biodiesel production, the green alga Chlorella protothecoides can efficiently produce oleaginous biomass and the lipid biosynthesis is greatly influenced by the availability of nitrogen source and corresponding nitrogen assimilation pathways. Based on isotope‐assisted kinetic flux profiling (KFP), the fluxes through the nitrogen utilization pathway were quantitatively analyzed. We found that autotrophic C. protothecoides cells absorbed ammonium mainly through glutamate dehydrogenase (GDH), and partially through glutamine synthetase (GS), which was the rate‐limiting enzyme of nitrogen assimilation process with rare metabolic activity of glutamine oxoglutarate aminotransferase (GOGAT, also known as glutamate synthase); whereas under heterotrophic conditions, the cells adapted to GS‐GOGAT cycle for nitrogen assimilation in which GS reaction rate was associated with GOGAT activity. The fact that C. protothecoides chooses the adenosine triphosphate‐free and less ammonium‐affinity GDH pathway, or alternatively the energy‐consuming GS‐GOGAT cycle with high ammonium affinity for nitrogen assimilation, highlights the metabolic adaptability of C. protothecoides exposed to altered nitrogen conditions.     

High temperature effects on ammonium assimilation in leaves of two Festuca arundinacea cultivars with different heat susceptibility

The aim of this study was to determine the effects of high temperature stress on ammonium assimilation in leaves of two tall fescue cultivars (Festuca arundinacea), Jaguar 3 brand (J3) (heat-tolerant) and TF 66 (T6) (heat-sensitive). High temperature stress for either 10 d or 20 d, and particularly the 20 d stress, produced dramatic changes in ammonium assimilation. After 20 d of stress treatment, the accumulations of total nitrogen, nitrate, soluble protein and total free amino acid (20 amino acids) decreased in both cultivars. Moreover, the activities of main regulatory enzymes, such as nitrate reductase, glutamine synthetase (GS), NADH-dependent glutamate synthase (GOGAT), as well as Δ¹-pyrroline-5-carboxylate reductase (P5CR), also decreased in both cultivars when exposed to 20 d stress. Heat stress had little influence on ammonium accumulation in J3, but this was not the case with T6. The accumulations of nitrate, ammonium, soluble protein, and total free amino acid between the two cultivars were different. This suggests that accumulations of these nitrogen forms were associated with heat tolerance in both tall fescue cultivars. Changes of both NADH-glutamate dehydrogenase (NADH-GDH) activity and Glx (glutamine and glutamic acid) concentration in both cultivars indicated that there is an alternative system for assimilation of nitrogen through glutamate dehydrogenase (GDH) in T6 during longer high temperature stress periods. Our results provide an insight to further selection and breeding of heat-tolerant tall fescue turfgrass cultivars.     

Glutamine synthetase and glutamate synthase from Sclerotinia sclerotiorum

Glutamine synthetase (GS, EC 6.3.1.2) and glutamate synthase (GOGAT, EC 1.4.1.13) were purified from Sclerotinia sclerotiorum and some of their properties studied. The GS transferase and biosynthetic activities, as well as GOGAT activity, were sensitive to feedback inhibition by amino acids and other metabolites. GS showed a marked dependence on ADP in the transferase reaction and on ATP in the Mg²⁺-dependent biosynthetic reaction. Regulation of GS activity by adenylylation/deadenylylation was demonstrated by snake venom phosphodiesterase treatment of the purified enzyme. GOGAT required NADPH as an electron donor; NADH was inactive. GOGAT was strongly inhibited by p-chloromercuribenzoate and the inhibition was reversed by cysteine. The enzyme was also markedly inhibited by o-phenanthroline, 2,2′-bipyridyl and azaserine. l-Methionine-dl-sulphoximine (MSX) and azaserine inhibited the incorporation of ¹⁵N-labelled ammonium sulphate into washed cells of S. sclerotiorum. MSX and azaserine respectively also inhibited purified GS and GOGAT activities. GDH activity was not detected in cell-extracts. Thus the GS/GOGAT pathway is the main route for the assimilation of ammonium compounds in this fungus.     

Field investigations of ammonia exchange between barley plants and the atmosphere. II. Nitrogen realiocation, free ammonium content and activities of ammonium-assimilating enzymes in different leaves

The activities of glutamine synthetase (GS) and glutamate synthase (GOGAT) in different leaves of field-grown spring barley were measured during the reproductive growth phase in 2 consecutive years. Concurrently, the contents of soluble ammonium ions and free amides in the leaves were determined. The studies were carried out to investigate the relationship between variations in these parameters and emission of NH3 from the plant foliage. GS and GOGAT activities declined very rapidly with leafage. The decline in enzyme activities was followed by an increase in soluble ammonium ions and amides in the leaf tissues. During the same period, about 75% of leaf and stem nitrogen was reallocated to the developing ear. The amount of NH₃ volatilized from the foliage during the reproductive growth phase amounted to about 1% of the reallocated nitrogen. The experimental years were characterized by very favourable conditions for grain dry matter formation and for re-utilization of nitrogen mobilized from leaves and stems. Ammonia volatilization occurring under conditions with declining GS and GOGAT activities and increasing tissue concentrations of NH₄⁺ may be useful in protecting the plant from accumulation of toxic NH₃ and NH₄⁺ concentrations in the tissues.     

Ammonia Assimilation in Canavalia ensiformis Plants under Water and Salt Stress

Nitrogen fixation and ammonia assimilation in nodules have beenthoroughly studied under stress conditions, but the behaviorof enzymes involved in ammonia assimilation to organic compoundsin plants of the Leguminosae family subjected to stress stillremains to be conclusively established. We found that understress conditions, C. ensiformis plants can switch from theirusual pathway of assimilation to an alternative one dependingon the nature of the stress and the tissue in which the processtakes place. In roots, it switches from the glutamate dehydrogenase(GDH) pathway to the glutamine synthetase (GS)/glutamate synthase(GOGAT) cycle under water stress but not under salt stress.However, in leaves under salt stress, GDH activity is maintainedbut GS activity markedly decreases (Received March 24, 1987; Accepted March 4, 1988)