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1.
Glutamine synthetase (GS), glutamate synthase (GOGAT), glutamate dehydrogenase (GDH), alanine dehydrogenase (ADH) and alanine aminotransferase (GPT) were detected in the cell-free homogenate ofStreptomyces avermitilis grown in a defined medium containing ammonium sulfate as the only nitrogen source. At an initial NH4 + concentration of 7.5 mmol/L, high activities of GS, GOGAT and GDH were found while that of ADH was low. The ADH activity was markedly increased at initially millimolar NH4 + concentrations. In some characteristics of its NH4 +-assimilating system (e.g. control of some enzyme activities, the NADPH specificity of GOGAT, the presence of alanine aminotransferase),S. avermitilis differs from other known streptomycetes.  相似文献   

2.
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.  相似文献   

3.
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.  相似文献   

4.
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.  相似文献   

5.
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.  相似文献   

6.
Ammonium ion accumulation in detached rice leaves treated with phosphinothricin (PPT), an inhibitior of glutamine synthetase (GS), was investigated in the light and darkness. PPT treatment increased NH4 + content and induced toxicity in rice leaves in the light but not in darkness, suggesting the importance of light in PPT-induced NH4 + toxicity in detached rice leaves. PPT treatment in the light resulted in a decrease of activities of the cytosolic form of GS and the chloroplastic form of GS. The photosynthetic electron transport inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea reduced NH4 + accumulation induced by PPT in the light. In darkness, PPT-induced NH4 + accumulation and toxicity were observed in the presence of glucose or sucrose.  相似文献   

7.
Alanine dehydrogenase in Arthrobacter fluorescens exhibited an allosteric behaviour and two K m values for ammonium were estimated. In batch cultures at different ammonium concentrations and in continuous culture following an NH4 + pulse, the level of ADH activity seems to be regulated by the ammonium concentration, high activities being observed when extracellular ammonium was in excess. The response to the growth rate of an ammonium-limited chemostat culture of A. fluorescens seems to indicate that alanine dehydrogenase and glutamine synthetase activities were inversely related. High activities of glutamate oxaloacetate transaminase and glutamate pyruvate transaminase have been found in crude extract of ammonium-limited cultures. From the results obtained in batch cultures grown at different glucose concentrations and in carbon-limited chemostat culture it appeared that the limitation by glucose influenced alanine dehydrogenase activity negatively. No glutamate dehydrogenase activity and no glutamate synthase activity could be detected with either NADH or NADPH as coenzymes.Abbreviations ADH alanine dehydrogenase - GS glutamine synthetase - GDH glutamate dehydrogenase - GOGAT glutamine oxoglutarate aminotransferase - GOT glutamate oxaloacetate transaminase - GPT glutamate pyruvate transaminase  相似文献   

8.
9.
Responses to excessive ammonium (NH4 +) were compared between two Arabidopsis ecotypes (Col-0, JA22) with respect to different photoperiods in hydroponics. In this study, we showed that external extra NH4 + led to severe growth suppression, accumulations of free NH4 + and amino acids and increased the activities of glutamine synthetase (GS) and glutamate dehydrogenase (GDH) in shoots of the two Arabidopsis ecotypes. However, the levels of free NH4 + and total amino acids increased, whereas the activities of GS, NADH-dependent glutamate synthase and GDH decreased under the continuous light when compared with the light (16 h)–dark (8 h) cycle photoperiod. Statistical analyses suggested that strong correlations exist among the growth reduction, accumulations of free NH4 +, total amino acids and levels of GS activity in shoots under the high NH4 + stress regardless of the photoperiod regimes. Interestingly, under the continuous light, Col-0 showed more resistant to such growth reduction and maintained about onefold higher capability of converting excess free NH4 + into amino acids, with onefold higher GS activity induced by the external NH4 + when compared with JA22. In contrast, these differences were abolished between Col-0 and JA22 under the light–dark cycle condition. Taken together, our results conclude that the sensitivity to NH4 + of Col-0 and JA22 is changed between the continuous light and the light–dark cycle photoperiod, which is correlative to the alteration of the GS activity in shoots.  相似文献   

10.
Cellular compartmentation of ammonium assimilation in rice and barley   总被引:9,自引:0,他引:9  
This review describes immunolocalization studies of the tissue and cellular location of glutamine synthetase (GS; EC 6.3.1.2) and glutamate synthase (Fd GOGAT; EC 1.4.7.1 and NADH-GOGAT; EC 1.4.1.14) proteins in roots and leaves of rice (Oryza sativa L.) and barley (Hordeum vulgare L.). In rice, cytosolic GS (GS1) protein was distributed homogeneously through all cells of the root. NADH GOGAT protein was strongly induced and its cellular location altered by ammonium treatment, becoming concentrated within the epidermal and exodermal cells. Fd GOGAT protein location changed with root development, from a widespread distribution in young cells to becoming concentrated within the central cylinder as cells matured. Plastid GS protein was barely detectable in rice roots, but was the major isoform in leaves, being present in the mesophyll and parenchyma sheath cells. GS1 was specific to the vascular bundle, as was NADH GOGAT, whereas Fd GOGAT was primarily found in mesophyll cells. In barley roots, GS1 protein was found in the cortical and vascular parenchyma and its concentration was highest in N-deficient seedlings. Plastid GS protein was detected in both cortical and vascular cells, where different plastid forms, containing different concentrations of GS protein, were identified. In barley leaves, GS2 protein was detected in the mesophyll chloroplasts and GS1 was found in the mesophyll and vascular cells. N nutrition strongly influenced this distribution, with a marked increase in GS1 concentration in the vascular cells in response to nitrate and ammonium, and an increase in mesophyll GS2 concentration in nitrate-grown seedlings. Fd GOGAT protein was found in both the mesophyll and vascular plastids. These localization studies show that the GS/GOGAT cycle is highly compartmentalized at both the subcellular and cellular levels. Reasons for this compartmentation, and the roles of each isoform, are discussed.  相似文献   

11.
The role of H2O2 in abscisic acid (ABA)-induced NH4+ accumulation in rice leaves was investigated. ABA treatment resulted in an accumulation of NH4+ in rice leaves, which was preceded by a decrease in the activity of glutamine synthetase (GS) and an increase in the specific activities of protease and phenylalanine ammonia-lyase (PAL). GS, PAL, and protease seem to be the enzymes responsible for the accumulation of NH4+ in ABA-treated rice leaves. Dimethylthiourea (DMTU), a chemical trap for H2O2, was observed to be effective in inhibiting ABA-induced accumulation of NH4+ in rice leaves. Inhibitors of NADPH oxidase, diphenyleneiodonium chloride (DPI) and imidazole (IMD), and nitric oxide donor (N-tert-butyl-α-phenylnitrone, PBN), which have previously been shown to prevent ABA-induced increase in H2O2 contents in rice leaves, inhibited ABA-induced increase in the content of NH4+. Similarly, the changes of enzymes responsible for NH4+ accumulation induced by ABA were observed to be inhibited by DMTU, DPI, IMD, and PBN. Exogenous application of H2O2 was found to increase NH4+ content, decrease GS activity, and increase protease and PAL-specific activities in rice leaves. Our results suggest that H2O2 is involved in ABA-induced NH4+ accumulation in rice leaves.  相似文献   

12.
Nitrate (NO3) and ammonium (NH4+) are the main forms of nitrogen available in the soil for plants. Excessive NH4+ accumulation in tissues is toxic for plants and exclusive NH4+-based nutrition enhances this effect. Ammonium toxicity syndrome commonly includes growth impairment, ion imbalance and chlorosis among others. In this work, we observed high intraspecific variability in chlorophyll content in 47 Arabidopsis thaliana natural accessions grown under 1 mM NH4+ or 1 mM NO3 as N-source. Interestingly, chlorophyll content increased in every accession upon ammonium nutrition. Moreover, this increase was independent of ammonium tolerance capacity. Thus, chlorosis seems to be an exclusive effect of severe ammonium toxicity while mild ammonium stress induces chlorophyll accumulation.  相似文献   

13.
The effect of potentially toxic concentrations of ammonium on root development of Scots pine seedlings raised on Perlite was investigated during growth periods of 3 or 10 weeks after sowing. It was shown that imbalanced ammonium nutrition led to conspicuous changes of root morphology provided the pH value in the medium was allowed to decrease to 3.9 due to the NH+4-dependent proton excretion into the rhizosphere. Ammonium toxicity could not be observed with seedlings treated either with ammonium nitrate or with ammonium chloride at pH 5.3 ? 6.8. While the supply of NH+4 considerably inhibited root development the biomass production of the shoot was increased. Determination of the endogenous level of ammonium in roots and the leaf whorl exclude a simple causal correlation between ammonium toxicity and accumulated ammonium as has been postulated for herbaceous plants.  相似文献   

14.
Li G  Dong G  Li B  Li Q  Kronzucker HJ  Shi W 《Planta》2012,235(2):239-252
Ammonium (NH4 +) toxicity is a significant agricultural problem globally, compromising crop growth and productivity in many areas. However, the molecular mechanisms of NH4 + toxicity are still poorly understood, in part due to a lack of valuable genetic resources. Here, a novel Arabidopsis mutant, amos2 (ammonium overly sensitive 2), displaying hypersensitivity to NH4 + in both shoots and roots, was isolated. The mutant exhibits the hallmarks of NH4 + toxicity at significantly elevated levels: severely suppressed shoot biomass, increased leaf chlorosis, and inhibition of lateral root formation. Amos2 hypersensitivity is associated with excessive NH4 + accumulation in shoots and a reduction in tissue potassium (K+), calcium (Ca2+), and magnesium (Mg2+). We show that the lesion is specific to the NH4 + ion, is independent of NH4 + metabolism, and can be partially rescued by elevated external K+. The amos2 lesion was mapped to a 16-cM interval on top of chromosome 1, where no similar mutation has been previously mapped. Our study identifies a novel locus controlling cation homeostasis under NH4 + stress and provides a tool for the future identification of critical genes involved in the development of NH4 + toxicity.  相似文献   

15.
Enzymatic activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS) participating in the nitrogen metabolism and related ammonium absorption were assayed after the microalga Chlorella vulgaris Beij. was jointly immobilized with the microalgae‐growth‐promoting bacterium Azospirillum brasilense. At initial concentrations of 3, 6, and 10 mg · L?1 NH4+, joint immobilization enhances growth of C. vulgaris but does not affect ammonium absorption capacity of the microalga. However, at 8 mg · L?1 NH4+, joint immobilization enhanced ammonium absorption by the microalga without affecting the growth of the microalgal population. Correlations between absorption of ammonium per cell and per culture showed direct (negative and positive) linear correlations between these parameters and microalga populations at 3, 6, and 10 mg · L?1 NH4+, but not at 8 mg · L?1 NH4+, where the highest absorption of ammonium occurred. In all cultures, immobilized and jointly immobilized, having the four initial ammonium concentrations, enzymatic activities of Chlorella are affected by A. brasilense. Regardless of the initial concentration of ammonium, GS activity in C. vulgaris was always higher when jointly immobilized and determined on a per‐cell basis. When jointly immobilized, only at an initial concentration of 8 mg · L?1 NH4+ was GDH activity per cell higher.  相似文献   

16.
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.  相似文献   

17.
The effects of NaCl on changes in ammonium level and enzyme activities of ammonium assimilation in roots growth of rice (Oryza sativa L.) seedlings were investigated. NaCl was effective in inhibiting root growth and stimulated the accumulation of ammonium in roots. Accumulation of ammonium in roots preceded inhibition of root growth caused by NaCl. Both effects caused by NaCl are reversible. Exogenous ammonium chloride and methionine sulfoximine (MSO), which caused ammonium accumulation in roots, inhibited root growth of rice seedlings. NaCl decreased glutamine synthetase and glutamate synthase activities in roots, but increased glutamate dehydrogenase activity. The growth inhibition of roots by NaCl or MSO could be reversed by the addition of L-glutamic acid or L-glutamine. The current results suggest that disturbance of ammonium assimilation in roots may be involved in regulating root growth reduction caused by NaCl.Abbreviations GDH glutamate dehydrogenase - GOGAT glutamate synthase - GS glutamine synthetase - MSO methionine sulfoximine  相似文献   

18.
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.  相似文献   

19.
The phytotoxicity of aluminium (Al) ions can be alleviated by ammonium (NH4+) in rice and this effect has been attributed to the decreased Al accumulation in the roots. Here, the effects of different nitrogen forms on cell wall properties were compared in two rice cultivars differing in Al tolerance. An in vitro Al‐binding assay revealed that neither NH4+ nor NO3? altered the Al‐binding capacity of cell walls, which were extracted from plants not previously exposed to N sources. However, cell walls extracted from NH4+‐supplied roots displayed lower Al‐binding capacity than those from NO3?‐supplied roots when grown in non‐buffered solutions. Fourier‐transform infrared microspectroscopy analysis revealed that, compared with NO3?‐supplied roots, NH4+‐supplied roots possessed fewer Al‐binding groups (‐OH and COO‐) and lower contents of pectin and hemicellulose. However, when grown in pH‐buffered solutions, these differences in the cell wall properties were not observed. Further analysis showed that the Al‐binding capacity and properties of cell walls were also altered by pHs alone. Taken together, our results indicate that the NH4+‐reduced Al accumulation was attributed to the altered cell wall properties triggered by pH decrease due to NH4+ uptake rather than direct competition for the cell wall binding sites between Al3+ and NH4+.  相似文献   

20.
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。  相似文献   

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