NADH-dependent Glutamate Synthase Research Articles

Genetic stability of Mycobacterium abscessus during antibiotic treatment

Genetic changes in Mycobacterium abscessus during antibiotic treatment are not fully understood. This study aimed to investigate the genetic changes in M. abscessus in patients receiving antibiotic treatment, and their clinical implications. Pretreatment and 12-month post-treatment M. abscessus isolates were obtained from patients with M. abscessus pulmonary disease. Isolates from each time point were separated into six groups based on their distinctive morphological characteristics. Twenty-four isolates, comprising 12 from patient A exhibiting progressive disease and 12 from patient B demonstrating stable disease, underwent sequencing. Subsequently, minimal inhibitory concentrations (MICs) for the administered antibiotics were measured. Persistent infection with a single strain was observed in patients A and B. During 12 months of treatment, MICs for administered drugs did not generally change over time in either patient and single nucleotide variations (SNV) associated with antimicrobial resistance (rrl, rrs, erm(41), gyrA, gyrB, whiB7 and hflX) were not mutated. Although not significant, 47 and 52 non-synonymous SNVs occurred in M. abscessus from patients A and B, respectively, and the accumulation of these SNVs differed in patients A and B, except for five SNVs. The most variable positions were within a probable NADH-dependent glutamate synthase gene and a putative YrbE family protein gene in patients A and B, respectively. Persistent infections by a single strain of M. abscessus were observed in two patients with different clinical courses. Genetic changes in M. abscessus during antibiotic treatment were relatively stable in these patients. NCT01616745 (ClinicalTrials.gov ID).

Coregulation of glutamine synthetase1;2 (GLN1;2) and NADH-dependent glutamate synthase (GLT1) gene expression in Arabidopsis roots in response to ammonium supply.

Ammonium absorbed by roots is assimilated into amino acids. The glutamine synthetase/glutamate synthase (glutamine 2-oxoglutarate aminotransferase) (GS/GOGAT) cycle is essential to this biological process. In Arabidopsis thaliana, GLN1;2 and GLT1 are the GS and GOGAT isoenzymes induced in response to ammonium supply and playing key roles in ammonium utilization. Although recent studies suggest gene regulatory networks involved in transcriptional regulation of ammonium-responsive genes, direct regulatory mechanisms for ammonium-induced expression of GS/GOGAT remain unclear. In this study, we revealed that the expression of GLN1;2 and GLT1 in Arabidopsis is not directly induced by ammonium but is regulated by glutamine or post-glutamine metabolites produced by ammonium assimilation. Previously, we identified a promoter region required for ammonium-responsive expression of GLN1;2. In this study, we further dissected the ammonium-responsive region of the GLN1;2 promoter and also performed a deletion analysis of the GLT1 promoter, which led to the identification of a conserved ammonium-responsive region. Yeast one-hybrid screening using the ammonium-responsive region of the GLN1;2 promoter as a decoy sequence revealed a trihelix family transcription factor DF1 that binds to this region. A putative DF1 binding site was also found in the ammonium-responsive region of the GLT1 promoter.

Inoculation of wheat with Bacillus sp. wp-6 altered amino acid and flavonoid metabolism and promoted plant growth.

Inoculation of wheat seedling with Bacillus sp. wp-6 changed amino acid metabolism and flavonoid synthesis and promoted plant growth. Plant growth-promoting rhizobacteria (PGPR), which can reduce the use of agrochemicals, is vital for the development of sustainable agriculture. In this study, proteomics and metabolomics analyses were performed to investigate the effects of inoculation with a PGPR, Bacillus sp. wp-6, on wheat (Triticum aestivum L.) seedling growth. The results showed that inoculation with Bacillus sp. wp-6 increased shoot and root fresh weights by 19% and 18%, respectively, after 40days. The expression levels of alpha-linolenic acid metabolism-related proteins and metabolites (lipoxygenase 2, allene oxide synthase 2, jasmonic acid, 17-hydroxylinolenic acid) and flavonoid biosynthesis-related proteins and metabolites (chalcone synthase 2 and PHC 4'-O-glucoside) were up-regulated. In addition, the expression levels of amino acid metabolism-related proteins (NADH-dependent glutamate synthase, bifunctional aspartokinase/homoserine, anthranilate synthase alpha subunit 1, and 3-phosphoshikimate 1-carboxyvinyltransferase) and metabolites (L-aspartate, L-arginine, and S-glutathionyl-L-cysteine) were also significantly up-regulated. Among them, NADH-dependent glutamate synthase and bifunctional aspartokinase/homoserine could act as regulators of nitrogen metabolism. Overall, inoculation of wheat with Bacillus sp. wp-6 altered alpha-linolenic acid metabolism, amino acid metabolism, and flavonoid synthesis and promoted wheat seedling growth. This study will deepen our understanding of the mechanism by which Bacillus sp. wp-6 promotes wheat growth using proteomics and metabolomics.

The Dynamics of NO3- and NH4+ Uptake in Duckweed Are Coordinated with the Expression of Major Nitrogen Assimilation Genes.

Duckweed plants play important roles in aquatic ecosystems worldwide. They rapidly accumulate biomass and have potential uses in bioremediation of water polluted by fertilizer runoff or other chemicals. Here we studied the assimilation of two major sources of inorganic nitrogen, nitrate () and ammonium (), in six duckweed species: Spirodela polyrhiza, Landoltia punctata, Lemna aequinoctialis, Lemna turionifera, Lemna minor, and Wolffia globosa. All six duckweed species preferred over and started using only when was depleted. Using the available genome sequence, we analyzed the molecular structure and expression of eight key nitrogen assimilation genes in S. polyrhiza. The expression of genes encoding nitrate reductase and nitrite reductase increased about 10-fold when was supplied and decreased when was supplied. and induced the glutamine synthetase (GS) genes GS1;2 and the GS2 by 2- to 5-fold, respectively, but repressed GS1;1 and GS1;3. and upregulated the genes encoding ferredoxin- and NADH-dependent glutamate synthases (Fd-GOGAT and NADH-GOGAT). A survey of nitrogen assimilation gene promoters suggested complex regulation, with major roles for NRE-like and GAATC/GATTC cis-elements, TATA-based enhancers, repeats, and G-quadruplex structures. These results will inform efforts to improve bioremediation and nitrogen use efficiency.

The Papain-like Cysteine Protease HpXBCP3 from Haematococcus pluvialis Involved in the Regulation of Growth, Salt Stress Tolerance and Chlorophyll Synthesis in Microalgae.

The papain-like cysteine proteases (PLCPs), the most important group of cysteine proteases, have been reported to participate in the regulation of growth, senescence, and abiotic stresses in plants. However, the functions of PLCPs and their roles in stress response in microalgae was rarely reported. The responses to different abiotic stresses in Haematococcus pluvialis were often observed, including growth regulation and astaxanthin accumulation. In this study, the cDNA of HpXBCP3 containing 1515 bp open reading frame (ORF) was firstly cloned from H. pluvialis by RT-PCR. The analysis of protein domains and molecular evolution showed that HpXBCP3 was closely related to AtXBCP3 from Arabidopsis. The expression pattern analysis revealed that it significantly responds to NaCl stress in H. pluvialis. Subsequently, transformants expressing HpXBCP3 in Chlamydomonas reinhardtii were obtained and subjected to transcriptomic analysis. Results showed that HpXBCP3 might affect the cell cycle regulation and DNA replication in transgenic Chlamydomonas, resulting in abnormal growth of transformants. Moreover, the expression of HpXBCP3 might increase the sensitivity to NaCl stress by regulating ubiquitin and the expression of WD40 proteins in microalgae. Furthermore, the expression of HpXBCP3 might improve chlorophyll content by up-regulating the expression of NADH-dependent glutamate synthases in C. reinhardtii. This study indicated for the first time that HpXBCP3 was involved in the regulation of cell growth, salt stress response, and chlorophyll synthesis in microalgae. Results in this study might enrich the understanding of PLCPs in microalgae and provide a novel perspective for studying the mechanism of environmental stress responses in H. pluvialis.

Effect of a mix of oligogalacturonides on symbiotic nitrogen fixation in common bean

The objective of this study was to evaluate the effect of a mix of pectic oligosaccharides in the common bean-rhizobia symbiosis. Cuba Cueto-25-9-N bean seeds were inoculated with Rhizobium tropici CIAT 899 and treated with a mix of oligogalacturonides, either by application to the seed (10 mg L-1, 1 ml per seed) at sowing and inoculation or by foliar spray (5 or 100 mg L-1, 1.5 ml per plant) to plants with two trefoils. Plant growth, nodulation, nitrogenase activity, and the gene expression of glutamine synthetase and glutamate synthase in nodules were evaluated in inoculated plants and oligogalacturonide-treated inoculated plants at 18 d post-inoculation. The oligogalacturonide-treated plants showed increases in shoot and root growth and number of nodules. Regarding nodule function, the oligogalacturonides increased the nitrogenase activity and the expression level of the gene coding for NADH-dependent glutamate synthase. The positive effects of oligogalacturonides resulted in higher effectiveness of the symbiotic nitrogen fixation. The application of oligogalacturonides can offer an alternative to increasing the symbiotic nitrogen fixation in common bean plants.

Effect of Dihydroartemisinin on Plasmodium NADH-Dependent Glutamate Synthase: The Implication in Malaria Management.

Artemisinin and its analogues (ARTs) are currently the most effective anti-malarial drugs, but the precise mechanism of action is still highly controversial. Effects of ARTs on Plasmodium genes expression are studied in our Lab. The overexpression of an interesting amidotransferase, NADH-dependent glutamate synthase (NADH-GltS) was found in treated by dihydroartemisinin (DHA). The increased expression occurred not only from global transcriptomics analysis on the human malaria parasite Plasmodium falciparum (P.falciparum) 3D7 and gene expression screening on all of iron-sulphur cluster proteins from . 3D7 invitro but also from Plasmodium berghei (P.berghei) ANKA in mice. Influence of DHA on NADH-GltS was specifically at trophozoite stage of P.falciparum and in a dose-dependent manner below the effective doses. L-glutamine (Gln) and L-glutamate (Glu) are the substrate and product of NADH-GltS respectively. Azaserine (Aza) is specific inhibitor for NADH-GltS. Experimental data showed that Glu levels were significantly decreasing with DHA dose increasing but NADH-GltS enzyme activities were still remained at higher levels in parasites, and appropriate amount of exogenous Glu could significantly reduce anti-malarial action of DHA but excessive amount lost the above effect. Aza alone could inhibit proliferation of P. falciparum and had an additive effect in combination with DHA. Those results could suggest that: Glutamate depletion is one of the anti-malarial actions of DHA; overexpression of NADH-GltS would be a feedback pattern of parasite itself due to glutamate depletion, but not a direct action of DHA; the "feedback pattern" is one of protective strategies of Plasmodium to interfere with the anti-malarial actions of DHA; and specific inhibitor for NADH-GltS as a new type of anti-malarial agents or new partner in ACT might provide a potential.

Uptake and utilization of different nitrogen forms in erect panicle japonica rice cultivar

ABSTRACT The response of rice to nitrogen (N) was compared between three rice cultivars and three N treatments: 1:0, 0:1, and 1:1 NH4 +:NO3 −. The three rice cultivars used were: Shennong1401 (SN1401, erect panicle rice cultivar) and Habataki and Sasanishiki (curved panicle rice cultivars). At the booting stage, higher gene expression and enzyme activity associated with N metabolism of rice in the including-NO3 − solutions effected the secondary assimilation of N metabolism, and ultimately increased grain development. During the tillering stage, lower NO3 − uptake rates and glutamine synthetase and NADH-dependent glutamate synthase activity in Habataki and Sasanishiki under the NO3 − treatments resulted in lower effective panicle number per plant. However, no significant differences were observed in the grain yield per plant of SN1401 between NH4 + and NO3 − treatments. Our results indicate that the erect panicle rice cultivar can grow better under the including-NO3 − treatment.

Benzothiadiazole and nitrogen source modify the nitrogen metabolism in cucumber infected with Pseudomonas syringae pv. lachrymans

The influence of nitrogen source and benzothiadiazole (BTH) pretreatment on nitrogen metabolism-related enzyme activities was studied in cucumber plants infected with Pseudomonas syringae pv. lachrymans (Psl). The plants were grown hydroponically in media containing nitrate and urea at different ratios: 100% NO3− (100N), 75% NO3−/25% urea (75N) and 50% NO3−/50% urea (50N) and pretreated with BTH prior to Psl inoculation.In the presence of urea in the nutritional medium the activities of most enzymes in the control plants decreased. Disease development was influenced by nitrogen nutrition and the weakest symptoms occurred in the 75N plants. The BTH-induced disease restriction was dependent on the type of nitrogen nutrition and was most visible in the 50N plants (about 2-fold reduction of infected leaf area compared to the plants not pretreated with BTH). The nitrogen metabolism was modified by BTH and this effect was stronger than that of urea. Irrespective of the type of nitrogen nutrition, in the BTH-treated plants, the activities of nitrate reductase, ferredoxin- and NADH-dependent glutamate synthase decreased whereas those of NADH-dependent glutamate dehydrogenase (NADH-GDH), aminotransferases and NADP-dependent isocitrate dehydrogenase were enhanced compared to the controls. After infection, in all nutritional variants, the strongest effect of BTH pretreatment was observed for NADH-GDH on the 2 day, when its activity increased by 85–256% compared to the infected non-pretreated plants. Our results concerning the glutamate-metabolizing enzymes suggest that BTH treatment combined with low dose of urea may have a positive influence on the nitrogen management leading to the inhibition of disease symptom progress.

Tos17 insertion in NADH-dependent glutamate synthase genes leads to an increase in grain protein content in rice

Nutrition from cereal grains sustains human life, and the protein content of rice grains is an important source for humans. Recent forward genetics studies pointed out the significant contribution of NADH-dependent glutamate synthase (NADH-GOGAT) to increased grain protein content in durum wheat. The aim of this work was to investigate the contribution of the two NADH-GOGAT enzymes in rice, NADH-GOGAT1, and NADH-GOGAT2, to rice grain protein content, using a reverse genetics approach. The grain protein content and free amino acid concentration were determined in Tos17 insertion mutants of the structural gene of each protein. We found a significant increase in protein content in the NADH-GOGAT2 mutant and an increase in free amino acid concentration in both the NADH-GOGAT1 and the NADH-GOGAT2 mutants. These results provide the first nutritional characterization of the NADH-GOGAT mutants. Rice containing increased protein and amino acid concentrations may have potential to contribute to improved rice nutrition. Since previous field analysis had indicated a 40% yield reduction in each of the mutants, future work should focus on improving the yield of the mutants.

Protein Hydrolysate Stimulates Growth in Tomato Coupled With N-Dependent Gene Expression Involved in N Assimilation.

Plant-derived protein hydrolysates (PHs) have received increased attention in the last decade because of their potential to improve yield, nutritional quality as well as tolerance to abiotic stressors. The current study investigated the effects and the molecular mechanisms of a legume-derived PH under optimal and sub-optimal nitrogen (N) concentrations (112 and 7 mg L−1, respectively) in tomato (Solanum lycopersicum L.). Growth and mineral composition of tomato plants treated with PHs by foliar spray or substrate drench were compared to untreated plants. In addition, the expression was determined of genes encoding ammonium and nitrate transporters and seven enzymes involved in N metabolism: nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase 1 (GS1), glutamine synthetase 2 (GS2), ferredoxin-dependent glutamate synthase (GLT), NADH-dependent glutamate synthase (GLS), and glutamate dehydrogenase (GDH). The root and total plant dry weight, SPAD index and leaf nitrogen content were higher by 21, 17, 7, and 6%, respectively, in plants treated by a substrate drench in comparison to untreated tomato plants, whereas foliar application of PH gave intermediate values. PH-treated plants grown with lower N availability showed reduced expression of NR and NiR as well as of nitrate and ammonium transporter transcripts in both leaf and root tissues in comparison with untreated plants; this was especially pronounced after application of PH by substrate drench. Conversely, the transcript level of an amino acid transporter gene was up-regulated in comparison with untreated plants. At high N regime, the transcript levels of the ammonium and amino acid transporters and also NR, NiR, and GLT were significantly up-regulated in root after PH foliar and substrate drench applications compared with untreated plants. An up-regulation was also observed for GS1, GS2, and GDH transcripts in leaf after substrate drench. These results highlighted the potential benefits of using legume PH in vegetable production systems to increase growth and N-nutritional status of plants.

Phenylalanine as a nitrogen source induces root growth and nitrogen-use efficiency in Populus × canescens.

To investigate the physiological responses of poplars to amino acids as sole nitrogen (N) sources, Populus × canescens (Ait.) Smith plants were supplied with one of three nitrogen fertilizers (NH4NO3, phenylalanine (Phe) or the mixture of NH4NO3 and Phe) in sand culture. A larger root system, and decreased leaf size and CO2 assimilation rate was observed in Phe- versus NH4NO3-treated poplars. Consistently, a greater root biomass and a decreased shoot growth were detected in Phe-supplied poplars. Decreased enzymatic activities of nitrate reductase (NR), glutamate synthase (GOGAT) and glutamate dehydrogenase (GDH) and elevated activities of nitrite reductase (NiR), phenylalanine ammonia lyase (PAL), glutamine synthetase (GS) and asparagine synthase (AS) were found in Phe-treated roots. Accordingly, reduced concentrations of NH4+, NO3- and total N, and enhanced N-use efficiencies (NUEs) were detected in Phe-supplied poplars. Moreover, the transcript levels of putative Phe transporters ANT1 and ANT3 were upregulated, and the mRNA levels of NR, glutamine synthetase 2 (GS2), NADH-dependent glutamate synthase (NADH-GOGAT), GDH and asparagine synthetase 2 (ASN2) were downexpressed in Phe-treated roots and/or leaves. The 15N-labeled Phe was mainly allocated in the roots and only a small amount of 15N-Phe was translocated to poplar aerial parts. These results indicate that poplar roots can acquire Phe as an N source to support plant growth and that Phe-induced NUEs in the poplars are probably associated with NH4+ re-utilization after Phe deamination and the carbon bonus simultaneously obtained during Phe uptake.

Adaptation to rhizosphere acidification is a necessary prerequisite for wheat (Triticum aestivum L.) seedling resistance to ammonium stress

Because soil acidification accompanies ammonium (NH4+) stress, the tolerance of higher plants to ammonium is associated with their adaptation to root medium acidification. However, the underlying mechanisms of this adaptation have not been fully elucidated. The objective of this study was thus to elucidate the effect of rhizosphere pH on NH4+ tolerance in different winter wheat cultivars (Triticum aestivum L.). Hydroponic experiments were carried out on two wheat cultivars: AK58 (an NH4+-sensitive cultivar) and XM25 (an NH4+-tolerant cultivar). Four pH levels resembling acidified (4.0, 5.0, 6.0 and 7.0) were tested and 5 mM NH4+ nitrogen (AN) was used as a stress treatment, with 5 mM nitrate nitrogen used as a control. The addition of AN led to a severe reduction in biomass and an increase in free NH4+, amino acids, and the activities of glutamine synthetase (GS) and glutamate dehydrogenase (GDH) in the shoots and roots of the two wheat cultivars. Further decreases in growth medium pH led to further increases in free NH4+, but decreases in total amino acids and the activities of GS and NADH-dependent glutamate synthase (NADH-GDH). However, there was less of an increase in free NH4+ and less of a reduction in the activities of GS and NADH-GDH in the cultivar XM25 compared with AK58. In addition, total soluble sugar content and the root-to-shoot soluble sugar ratio were also decreased by AN treatment, except in the shoots of XM25. Decreasing pH resulted in lower root-to-shoot soluble sugar ratios with greater reductions in the AK58 cultivar. These results indicate that wheat growth was inhibited significantly by the addition of NH4+ combined with low pH. Low medium pH reduced the capacity for nitrogen assimilation and interrupted carbohydrate transport between the shoot and root. The NH4+-tolerant cultivar XM25 was better adapted to low rhizosphere pH due to its increased capacity for assimilating NH4+ efficiently and thereby avoiding toxic levels of intracellular NH4+ at low medium pH.

NADH-dependent glutamate synthase participated in ammonium assimilation in Arabidopsis root

Higher plants have 2 GOGAT species, Fd-GOGAT and NADH-GOGAT. While Fd-GOGAT mainly assimilates ammonium in leaves, which is derived from photorespiration, the function of NADH-GOGAT, which is highly expressed in roots,1 needs to be elucidated. The aim of this study was to clarify the role of NADH-GOGAT in Arabidopsis roots. The supply of ammonium to the roots caused an accumulation of NADH-GOGAT, while Fd-GOGAT 1 and Fd-GOGAT 2 showed no response. A promoter–GUS fusion analysis and immunohistochemistry showed that NADH-GOGAT was located in non-green tissues like vascular bundles, shoot apical meristem, pollen, stigma, and roots. The localization of NADH-GOGAT and Fd-GOGAT was not overlapped. NADH-GOGAT T-DNA insertion lines showed a reduction of glutamate and biomass under normal CO2 conditions. These data emphasizes the importance of NADH-GOGAT in the ammonium assimilation of Arabidopsis roots.

NADH-dependent glutamate synthase plays a crucial role in assimilating ammonium in the Arabidopsis root

Plant roots under nitrogen deficient conditions with access to both ammonium and nitrate ions, will take up ammonium first. This preference for ammonium rather than nitrate emphasizes the importance of ammonium assimilation machinery in roots. Glutamine synthetase (GS) and glutamate synthase (GOGAT) catalyze the conversion of ammonium and 2-oxoglutarate to glutamine and glutamate. Higher plants have two GOGAT species, ferredoxin-dependent glutamate synthase (Fd-GOGAT) and nicotinamide adenine dinucleotide (NADH)-GOGAT. While Fd-GOGAT participates in the assimilation of ammonium, which is derived from photorespiration in leaves, NADH-GOGAT is highly expressed in roots and its importance needs to be elucidated. While ammonium as a minor nitrogen form in most soils is directly taken up, nitrate as the major nitrogen source needs to be converted to ammonium prior to uptake. The aim of this study was to investigate and quantify the contribution of NADH-GOGAT to the ammonium assimilation in Arabidopsis (Arabidopsis thaliana Columbia) roots. Quantitative real-time polymerase chain reaction (PCR) and protein gel blot analysis showed an accumulation of NADH-GOGAT in response to ammonium supplied to the roots. In addition the localization of NADH-GOGAT and Fd-GOGAT did not fully overlap. Promoter-β-glucuronidase (GUS) fusion analysis and immunohistochemistry showed that NADH-GOGAT was highly accumulated in non-green tissue like vascular bundles, shoot apical meristem, pollen, stigma and roots. Reverse genetic approaches suggested a reduction in glutamate production and biomass accumulation in NADH-GOGAT transfer DNA (T-DNA) insertion lines under normal CO2 condition. The data emphasize the importance of NADH-GOGAT in the ammonium assimilation in Arabidopsis roots.

The role of malate in the synthesis of glutamate in Pisum arvense roots

The in vivo and in vitro activities of NADH-dependent glutamate synthase in excised <em>Pisum arvense</em> roots increased several-fold under the influence of malate while pyruvate oxaloacctate. citrate and succinate inhibited this entyme. The plastids isolated from <em>Pisum arvense</em> root,. ahen incubated with glutamine and α-ketoglutarate, released glutamate into the medium Malate clearly stimulated this process. Albizziin (25 mM) completely reduced the presence of glutamate in the incubation mixture. These results indicate that reduced pyridine nucleotides arising in <em>P. arvense</em> root plastids during oxidation of malic acid may constitute the indispensable source of electrons for glutamic acid synthesis.

Regulation of N2 fixation and NO3−/NH4+ assimilation in nodulated and N-fertilized Phaseolus vulgaris L. exposed to high temperature stress

Legumes need large amounts of N to grow satisfactorily. Under low NO3− availability in the soil, many legumes meet their N requirements by N2 fixation in association with rhizobia. Both NO3− uptake and N2 fixation decrease as temperature exceeds optimal growth conditions, but the mechanisms of regulation of N2 fixation and NO3−/NH4+ assimilation under high temperature stress are not completely understood. We describe an experiment in which physiological mechanisms regulating N metabolism of common bean (Phaseolus vulgaris L.) are investigated in plants submitted to daily maximum temperatures of 28, 34 and 39°C. Common bean was grown in symbiosis with each of six rhizobial strains—belonging to four different species and varying in N2 fixation effectiveness—or fertilized with NO3− until flowering. Harvest measurements included the activities of shoot, stem and root NO3− reductase (NR), nodule glutamine synthetase (GS), NADH-dependent glutamate synthase (GOGAT), nitrogenase, phosphoenol pyruvate carboxylase (PEPcase), N-export rates by nodules and concentration of N compounds in the xylem sap. Higher temperatures inhibited N2 fixation resulting in lower proportion of ureide-N in nodules and xylem sap of nodulated plants in relation to amide-N and α-amino-N. Higher temperatures consistently reduced the activity of NR in leaves of N-fertilized plants. Higher temperatures also decreased N exported from nodules and activities of nitrogenase, GS, GOGAT and PEPcase. The rate of decreases varied in plants with different strains. Furthermore, the activities of GS and GOGAT were more strongly affected by high temperatures than the activity of nitrogenase. There was a remarkable increase in the concentration of NH4+-N and ureide-N in the nodules when GS and GOGAT activities decreased. Therefore, the results provide evidence that N2 fixation in common bean submitted to heat stress is limited by NH4+ assimilation via GS-GOGAT rather than by decreased activity of nitrogenase. Rhizobial effectiveness determined the degree of down-regulation of GS-GOGAT activity in nodule tissues.

Beneficial Role of Exogenous Spermidine on Nitrogen Metabolism in Tomato Seedlings Exposed to Saline-alkaline Stress

We studied the effects of exogenous spermidine (Spd) on plant growth and nitrogen metabolism in two cultivars of tomato (Solanum lycopersicum) that have differential sensitivity to mixed salinity-alkalinity stress: ‘Jinpeng Chaoguan’ (salt-tolerant) and ‘Zhongza No. 9’ (salt-sensitive). Seedling growth of both tomato cultivars was inhibited by salinity-alkalinity stress, but Spd treatment alleviated the growth reduction to some extent, especially in ‘Zhongza No. 9’. Exogenous Spd may help reduce stress-induced increases in free amino acids, ammonium (NH4+) contents, and NADH-dependent glutamate dehydrogenase (NADH-GDH) activities; depress stress-induced decreases in soluble protein and nitrate content; and depress nitrate reductase, nitrite reductase, glutamine synthetase (GS), NADH-dependent glutamate synthase (NADH-GOGAT), glutamate oxaloacetate transaminase (GOT), and glutamate pyruvate transaminase (GPT) activities, especially for ‘Zhongza No. 9’. Based on our results, we suggest that exogenous Spd promotes the assimilation of excess toxic NH4+ by coordinating and strengthening the synergistic action of NADH-GDH, GS/NADH-GOGAT, and transamination pathways, all during saline-alkaline stress. Subsequently, NH4+ and its related enzymes (GDH, GS, GOGAT, GOT, and GPT), in vivo, are maintained in a proper and balanced state to enable mitigation of stress-resulted damages. These results suggest that exogenous Spd treatment can relieve nitrogen metabolic disturbances caused by salinity-alkalinity stress and eventually promote plant growth.

Role of glutamate availability on cadmium induced changes of nitrogen and glutamate in tomato plants (Solanum lycopersicon)

Maximal glutamine synthetase (GS, EC 6.3.1.2) activity and free aminoacid accumulation occur together in the leaves and roots. GS and specific GS activities although declining in the stressed leaves. In the other hand, the decrease of GS activity in the roots is contrarily accompanied by an increase in specific GS activity in roots in treated plants. Maximal GS activity is correlated with the peak degradation of the free aminoacid and protein in the two organs. Western blots analysis from Cd treated plants shows a decrease in chloroplastic GS (GS2) and an increase in cytosolic GS (GS1) proteins. In the roots, free ami- noacids accumulated as NADH—dependent glutamate synthase (NADH-GOGAT, EC 2.6.1.5.3) and specific activities increase. Maximal leaf NADH—GOGAT activity occurs simultaneously with maximal leaf aminating glutamate dehydrogenase (NADH-GDH, EC 1.4.1.2) activity.

イネにおけるNADH依存性グルタミン酸合成酵素 (NADH-GOGAT) 1及び2遺伝子の破壊変異体の獲得とその表現型

イネにおけるNADH依存性グルタミン酸合成酵素 (NADH-GOGAT) 1及び2遺伝子の破壊変異体の獲得とその表現型