scholarly journals How do three cytosolic glutamine synthetase isozymes of wheat perform N assimilation and translocation?

2019 ◽  
Author(s):  
Yihao Wei ◽  
Xiaochun Wang ◽  
Zhiyong Zhang ◽  
Shuping Xiong ◽  
Yiming Zhang ◽  
...  
Keyword(s):  
Glutamine Synthetase ◽  
Nitrogen Assimilation ◽  
Root Tip ◽  
Kinetic Properties ◽  
Meristematic Zone ◽  
Ammonium Toxicity ◽  
Vascular Tissues ◽  
High Affinity ◽  
N Assimilation ◽  
Cytosolic Glutamine Synthetase

AbstractTo understand how the three cytosolic glutamine synthetase (GS1) isozymes of wheat (Triticum aestivum L., TaGS1) perform nitrogen assimilation and translocation, we studied the kinetic properties of TaGS1 isozymes, the effects of nitrogen on the expression and localization of TaGS1 isozymes with specific antibodies, and the nitrogen metabolism. The results showed TaGS1;1, the dominant TaGS1 isozyme, had a high affinity for substrates, and was widely localized in the mesophyll cells, root pericycle and root tip meristematic zone, suggesting it was the primary isozyme for N assimilation. TaGS1;2, with a high affinity for Glu, was activated by Gln, and was mainly localized in the around vascular tissues, indicating that TaGS1;2 catalyzed Gln synthesis in low Glu concentration, then the Gln returned to activate TaGS1;2, which may lead to the rapid accumulation of Gln around the vascular tissues. TaGS1;3 had low affinity for substrates but the highest Vmax among TaGS1, was mainly localized in the root tip meristematic zone; exogenous NH4+ could promote TaGS1;3 expressing, indicating that TaGS1;3 could rapidly assimilate NH4+ to relieve NH4+ toxicity. In conclusion, TaGS1;1, TaGS1;2 and TaGS1;3 have different role in N assimilation, Gln translocation and relieving ammonium toxicity, respectively, and synergistically perform nitrogen assimilation and translocation.HighlightThree cytosolic glutamine synthase isozymes of wheat have different role and synergistically perform nitrogen assimilation and translocation.

Studies on possible routes of ammonium assimilation in soybean root nodule bacteroids

1973 ◽  
Vol 19 (12) ◽  
pp. 1493-1499 ◽  
Author(s):  
Stanley D. Dunn ◽  
Robert V. Klucas
Keyword(s):  
Glutamine Synthetase ◽  
Nitrogen Assimilation ◽  
Reductive Amination ◽  
Root Nodule ◽  
Root Nodules ◽  
Ammonium Assimilation ◽  
Kinetic Properties ◽  
Transferase Activity ◽  
Soybean Root

Glutamine amide–2-oxoglutarate aminotransferase NAD+ oxidoreductase (GOGAT), glutamine synthetase (GS), glutamate dehydrogenase (GD), and alanine dehydrogenase (AD) were studied in soybean root nodules. GS, GOGAT, and AD were present in bacteroids at levels that could account for ammonium assimilation, but GD activity was quite low. The total activities of GS and GD were higher in the cytosol than in the bacteroids by factors of 20 and 7, respectively, whereas GOGAT was not detected in the cytosol. GS (transferase activity) was inhibited by alanine, CTP, glycine, and tryptophan at 5 mM but was relatively unaffected by asparagine, aspartic acid, CMP, glucosamine, and histidine at 5 mM. GOGAT activity was unaffected by ATP, ADP, 8-hydroxyquinoline, and 1,10-phenanthroline but was inhibited by EDTA, citrate, and parachloromercuribenzoate. GOGAT activity (reductive amination) was also inhibited 97% by preincubation with 10−4 M azaserine for 30 min but GD activity was inhibited only 13%. The apparent Km values for NH4+ by AD was 7.4 × 10−3 M and by GD was 7.3 × 10−2 M while for glutamine by GOGAT it was 9.3 × 10−5 M. Activities and kinetic properties for these enzymes may suggest potential routes of nitrogen assimilation in vivo.

scholarly journals Cytosolic Glutamine Synthetase Gln1;2 Is the Main Isozyme Contributing to GS1 Activity and Can Be Up-Regulated to Relieve Ammonium Toxicity

2016 ◽  
Vol 171 (3) ◽  
pp. 1921-1933 ◽  
Author(s):  
Miao Guan ◽  
Thomas C. de Bang ◽  
Carsten Pedersen ◽  
Jan K. Schjoerring
Keyword(s):  
Glutamine Synthetase ◽  
Ammonium Toxicity ◽  
Cytosolic Glutamine Synthetase

scholarly journals Nitrogen Regulating the Expression and Localization of Four Glutamine Synthetase Isoforms in Wheat (Triticum aestivum L.)

2020 ◽  
Vol 21 (17) ◽  
pp. 6299
Author(s):  
Yihao Wei ◽  
Xiaochun Wang ◽  
Zhiyong Zhang ◽  
Shuping Xiong ◽  
Xiaodan Meng ◽  
...  
Keyword(s):  
Glutamine Synthetase ◽  
Nitrogen Assimilation ◽  
Vascular Tissue ◽  
Mrna Level ◽  
Triticum Aestivum L ◽  
Kinetic Properties ◽  
High Catalytic Activity ◽  
Root Tips ◽  
Plant Nitrogen ◽  
Key Enzyme

Glutamine synthetase (GS), the key enzyme in plant nitrogen assimilation, is strictly regulated at multiple levels, but the most relevant reports focus on the mRNA level. Using specific antibodies as probes, the effects of nitrogen on the expression and localization of individual wheat GS (TaGS) isoforms were studied. In addition to TaGS2, TaGS1;1 with high affinity to substrate and TaGS1;3 with high catalytic activity were also localized in mesophyll, and may participate in cytoplasmic assimilation of ammonium (NH4+) released from photorespiration or absorbed by roots; TaGS1;2 was localized in xylem of leaves. In roots, although there were hundreds of times more TaGS1;1 than TaGS1;2 transcripts, the amount of TaGS1;1 subunit was not higher than that of TaGS1;2; NH4+ inhibited TaGS1;1 expression but stimulated TaGS1;3 expression. In root tips, nitrate stimulated TaGS1;1, TaGS1;3, and TaGS2 expression in meristem, while NH4+ promoted tissue differentiation and TaGS1;2 expression in endodermis and vascular tissue. Only TaGS1;2 was located in vascular tissue of leaves and roots, and was activated by glutamine, suggesting a role in nitrogen transport. TaGS1;3 was induced by NH4+ in root endodermis and mesophyll, suggesting a function in relieving NH4+ toxicity. Thus, TaGS isoforms play distinct roles in nitrogen assimilation for their different kinetic properties, tissue locations, and response to nitrogen regimes.

scholarly journals The wheat cytosolic glutamine synthetase GS1.1 modulates N assimilation and spike development by characterizing CRISPR-edited mutants

2020 ◽  
Author(s):  
Yazhou Wang ◽  
Wan Teng ◽  
Yanpeng Wang ◽  
Xiang Ouyang ◽  
He Xue ◽  
...  
Keyword(s):  
Glutamine Synthetase ◽  
Field Experiments ◽  
Inorganic Nitrogen ◽  
Grain Filling ◽  
N Availability ◽  
Wild Type ◽  
Spike Development ◽  
N Assimilation ◽  
Cytosolic Glutamine Synthetase ◽  
Ammonium Stress

AbstractGlutamine synthetase (GS) mediates the first step in the assimilation of inorganic nitrogen (N) into amino acids, however the function of GS encoding genes is not well understood in wheat (Triticum aestivum). We found that the cytosolic TaGS1.1 was the major transcripted GS1 gene and was up-regulated by low-N availability. CRISPR/Cas9 mediated genome editing was employed to develop two gs1.1 mutants with mutated TaGS1.1-6A, −6B, and -6D. Both mutants had lower grains per spike and grain yield per plant than the wild type under both low-N and high-N conditions in field experiments. In a hydroponic culture treated with different N resources, the two mutants was more sensitive to low-N stress than the wild type, but showed similar sensitivity to high ammonium stress with the wild type. The growth deficiency and impaired spike development were associated with the imbalance of N metabolites in the mutant plants. During grain filling, TaGS1.1 mutation reduced N translocation efficiency and delayed leaf N loss and grain N filling. Our results suggested that TaGS1.1 is important for N assimilation and remobilization, and required for wheat adaptation to N-limited conditions and spike development.HighlightThe wheat cytosolic glutamine synthetase TaGS1.1 is important for N assimilation and remobilization, and is required for wheat adaptation to low-N stress and spike development.

scholarly journals The importance of cytosolic glutamine synthetase in nitrogen assimilation and recycling

2009 ◽  
Vol 182 (3) ◽  
pp. 608-620 ◽  
Author(s):  
Stéphanie M. Bernard ◽  
Dimah Z. Habash
Keyword(s):  
Glutamine Synthetase ◽  
Nitrogen Assimilation ◽  
Cytosolic Glutamine Synthetase

scholarly journals Integrative Transcriptomic and Proteomic Analysis Reveals an Alternative Molecular Network of Glutamine Synthetase 2 Corresponding to Nitrogen Deficiency in Rice (Oryza sativa L.)

2021 ◽  
Vol 22 (14) ◽  
pp. 7674
Author(s):  
Ting Liang ◽  
Zhengqing Yuan ◽  
Lu Fu ◽  
Menghan Zhu ◽  
Xiaoyun Luo ◽  
...  
Keyword(s):  
Glutamine Synthetase ◽  
Oryza Sativa L ◽  
Alternative Pathway ◽  
Rice Variety ◽  
Nitrogen Deficiency ◽  
Primary Root ◽  
Phenotypic Characterization ◽  
Glutathione Metabolism ◽  
N Assimilation ◽  
N Deficiency

Nitrogen (N) is an essential nutrient for plant growth and development. The root system architecture is a highly regulated morphological system, which is sensitive to the availability of nutrients, such as N. Phenotypic characterization of roots from LY9348 (a rice variety with high nitrogen use efficiency (NUE)) treated with 0.725 mM NH4NO3 (1/4N) was remarkable, especially primary root (PR) elongation, which was the highest. A comprehensive analysis was performed for transcriptome and proteome profiling of LY9348 roots between 1/4N and 2.9 mM NH4NO3 (1N) treatments. The results indicated 3908 differential expression genes (DEGs; 2569 upregulated and 1339 downregulated) and 411 differential abundance proteins (DAPs; 192 upregulated and 219 downregulated). Among all DAPs in the proteome, glutamine synthetase (GS2), a chloroplastic ammonium assimilation protein, was the most upregulated protein identified. The unexpected concentration of GS2 from the shoot to the root in the 1/4N treatment indicated that the presence of an alternative pathway of N assimilation regulated by GS2 in LY9348 corresponded to the low N signal, which was supported by GS enzyme activity and glutamine/glutamate (Gln/Glu) contents analysis. In addition, N transporters (NRT2.1, NRT2.2, NRT2.3, NRT2.4, NAR2.1, AMT1.3, AMT1.2, and putative AMT3.3) and N assimilators (NR2, GS1;1, GS1;2, GS1;3, NADH-GOGAT2, and AS2) were significantly induced during the long-term N-deficiency response at the transcription level (14 days). Moreover, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis demonstrated that phenylpropanoid biosynthesis and glutathione metabolism were significantly modulated by N deficiency. Notably, many transcription factors and plant hormones were found to participate in root morphological adaptation. In conclusion, our study provides valuable information to further understand the response of rice roots to N-deficiency stress.

scholarly journals Some Kinetic Properties of Bacillus subtilis Glutamine Synthetase

1970 ◽  
Vol 245 (20) ◽  
pp. 5206-5213 ◽  
Author(s):  
Thomas F. Deuel ◽  
E.R. Stadtman
Keyword(s):  
Bacillus Subtilis ◽  
Glutamine Synthetase ◽  
Kinetic Properties

scholarly journals Chloroplast and cytosolic glutamine synthetase are encoded by homologous nuclear genes which are differentially expressed in vivo.

1988 ◽  
Vol 263 (20) ◽  
pp. 9651-9657
Author(s):  
S V Tingey ◽  
F Y Tsai ◽  
J W Edwards ◽  
E L Walker ◽  
G M Coruzzi
Keyword(s):  
Glutamine Synthetase ◽  
Nuclear Genes ◽  
Differentially Expressed ◽  
Cytosolic Glutamine Synthetase

scholarly journals Gating Competence of Constitutively Open CLC-0 Mutants Revealed by the Interaction with a Small Organic Inhibitor

2003 ◽  
Vol 122 (3) ◽  
pp. 295-306 ◽  
Author(s):  
Sonia Traverso ◽  
Laura Elia ◽  
Michael Pusch
Keyword(s):  
Chloride Channels ◽  
Bound State ◽  
Side Chain ◽  
Pore Channel ◽  
Kinetic Properties ◽  
Wild Type ◽  
High Affinity ◽  
Critical Residue ◽  
Fast Gating

Opening of CLC chloride channels is coupled to the translocation of the permeant anion. From the recent structure determination of bacterial CLC proteins in the closed and open configuration, a glutamate residue was hypothesized to form part of the Cl−-sensitive gate. The negatively charged side-chain of the glutamate was suggested to occlude the permeation pathway in the closed state, while opening of a single protopore of the double-pore channel would reflect mainly a movement of this side-chain toward the extracellular pore vestibule, with little rearrangement of the rest of the channel. Here we show that mutating this critical residue (Glu166) in the prototype Torpedo CLC-0 to alanine, serine, or lysine leads to constitutively open channels, whereas a mutation to aspartate strongly slowed down opening. Furthermore, we investigated the interaction of the small organic channel blocker p-chlorophenoxy-acetic acid (CPA) with the mutants E166A and E166S. Both mutants were strongly inhibited by CPA at negative voltages with a >200-fold larger affinity than for wild-type CLC-0 (apparent KD at −140 mV ∼4 μM). A three-state linear model with an open state, a low-affinity and a high-affinity CPA-bound state can quantitatively describe steady-state and kinetic properties of the CPA block. The parameters of the model and additional mutagenesis suggest that the high-affinity CPA-bound state is similar to the closed configuration of the protopore gate of wild-type CLC-0. In the E166A mutant the glutamate side chain that occludes the permeation pathway is absent. Thus, if gating consists only in movement of this side-chain the mutant E166A should not be able to assume a closed conformation. It may thus be that fast gating in CLC-0 is more complex than anticipated from the bacterial structures.

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