Regulation Of Nitrogen Metabolism Research Articles

Genome-Wide Identification and Expression Analysis of GATA Gene Family under Different Nitrogen Levels in Arachis hypogaea L.

Nitrogen, one of the essential elements, is a key determinant for improving peanut growth and yield. GATA zinc finger transcription factors have been found to be involved in regulation of nitrogen metabolism. However, a systematic characterization of the GATA gene family and patterns of their expression under different nitrogen levels remains elusive. In this study, a total of 45 GATA genes distributed among 17 chromosomes were identified in the peanut genome and classified into three subfamilies I, II and III with 26, 13 and 6 members, respectively, whose physicochemical characteristics, gene structures and conserved motifs were also analyzed. Furthermore, the optimal level of nitrogen fertilizer on the growth of peanut cultivar Yuhua 23 was determined by pod yield and value cost ratio from 2020 to 2022, and the results revealed that 150 kg hm−2 nitrogen was the best for cultivation of peanut Yuhua 23 because of its highest pod yield and relatively higher VCR of more than four. In addition, expression patterns of peanut GATA genes under different nitrogen levels were detected by real-time quantitative PCR and several GATA genes were significantly changed under a nitrogen level of 150 kg hm−2. Overall, the above results would be helpful for further understanding biological functions of the GATA gene family in cultivated peanut.

Synergistic mitigation of nickel toxicity in pepper (Capsicum annuum) by nitric oxide and thiourea via regulation of nitrogen metabolism and subcellular nickel distribution.

Nickel (Ni) contamination hinders plant growth and yield. Nitric oxide (NO) and thiourea (Thi) aid plant recovery from heavy metal damage, but their combined effects on pepper (Capsicum annuum ) plant tolerance to Ni stress need more study. Sodium nitroprusside (0.1mM, SNP) and 400mgL-1 Thi, alone and combined, were studied for their impact on pepper growth under Ni toxicity. Ni stress reduces chlorophyll, PSII efficiency and leaf water and sugar content. However, SNP and Thi alleviate these effects by increasing leaf water, proline and sugar content. It also increased the activities of superoxide dismutase, catalase, ascorbate peroxidase and peroxidase. Nickel stress lowered nitrogen assimilation enzymes (nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase and glutamate dehydrogenase) and protein content, but increased nitrate, ammonium and amino acid content. SNP and Thi enhanced nitrogen assimilation, increased protein content and improved pepper plant growth and physiological functions during Ni stress. The combined treatment reduced Ni accumulation, increased Ni in leaf cell walls and potentially in root vacuoles, and decreased Ni concentration in cell organelles. It effectively mitigated Ni toxicity to vital organelles, surpassing the effects of SNP or Thi use alone. This study provides valuable insights for addressing heavy metal contamination in agricultural soils and offers potential strategies for sustainable and eco-friendly farming practices.

Epitranscriptome changes triggered by ammonium nutrition regulate the proteome response of maritime pine roots.

Epitranscriptome constitutes a gene expression checkpoint in all living organisms. Nitrogen is an essential element for plant growth and development that influences gene expression at different levels such as epigenome, transcriptome, proteome, and metabolome. Therefore, our hypothesis is that changes in the epitranscriptome may regulate nitrogen metabolism. In this study, epitranscriptomic modifications caused by ammonium nutrition were monitored in maritime pine roots using Oxford Nanopore Technology. Transcriptomic responses mainly affected transcripts involved in nitrogen and carbon metabolism, defense, hormone synthesis/signaling, and translation. Global detection of epitranscriptomic marks was performed to evaluate this posttranscriptional mechanism in un/treated seedlings. Increased N6-methyladenosine (m6A) deposition in the 3'-UTR was observed in response to ammonium, which seems to be correlated with poly(A) lengths and changes in the relative abundance of the corresponding proteins. The results showed that m6A deposition and its dynamics seem to be important regulators of translation under ammonium nutrition. These findings suggest that protein translation is finely regulated through epitranscriptomic marks likely by changes in mRNA poly(A) length, transcript abundance and ribosome protein composition. An integration of multiomics data suggests that the epitranscriptome modulates responses to nutritional, developmental and environmental changes through buffering, filtering, and focusing the final products of gene expression.

Nitric Oxide- and Sulfur-Mediated Reversal of Cadmium-Inhibited Photosynthetic Performance Involves Hydrogen Sulfide and Regulation of Nitrogen, Sulfur, and Antioxidant Metabolism in Mustard

This study aimed to test the role of hydrogen sulfide (H2S) in the responses regarding the nitric oxide- (NO) and sulfur (S)-mediated improvement in photosynthesis and growth under cadmium (Cd) stress in mustard (Brassica juncea L. cv. Giriraj), and integrate the mechanisms of S, nitrogen (N), and antioxidant metabolism. The plants grown with Cd (200 mg Cd kg−1 soil) exhibited reduced assimilation of S and N and diminished photosynthetic performance, which was associated with higher Cd accumulation-induced excess reactive oxygen species (ROS) production. The application of 100 μM of sodium nitroprusside (SNP, a NO donor) together with a more prominent concentration of S resulted in increased photosynthetic S- and N-use efficiency, production of non-protein thiols and phytochelatins, efficiency of enzymatic (superoxide dismutase, ascorbate peroxidase, and glutathione reductase), non-enzymatic antioxidants (ascorbate and glutathione) limiting Cd accumulation and, thus, reduced oxidative stress (superoxide radical, hydrogen peroxide, and thiobarbituric acid reactive species content). The benefit of NO together with S was manifested through a modulation in H2S production. The use of 100 μM of hypotaurine (HT; H2S scavenger) or 100 μM of cPTIO (2-4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) in plants treated with NO plus S reversed the action of NO plus S, with a higher reduction in photosynthesis and growth with the use of HT, suggesting that H2S plays a significant role in the NO- and S-mediated alleviation of Cd stress. The interplay of NO and ES with H2S may be used in augmenting the photosynthesis and growth of Cd-grown mustard plants.

Could nitrogen compounds be indicators of tolerance to high doses of Cu and Fe in the cultivation of Leucaena leucocephala?

Nitrogen metabolism and the production of primary and secondary metabolites vary according to biotic and abiotic factors such as trace elements (TE) stress, and can, therefore, be considered biomarkers. The present study evaluated the effect of copper (Cu) and iron (Fe) TE, separately, on the metabolism of nitrogen compounds and biomass production, partitioned into shoot and roots of Leucaena leucocephala (Lam.) de Wit., and identified possible defense mechanisms linked to nitrogen metabolism. At 120 days of cultivation, the biomass production of L. leucocephala was higher when exposed to excess Fe than Cu. Nonetheless, the biomass gain (%) of plants exposed to Cu was higher, especially the biomass gains in roots. The tolerance and biomass production of L. leucocephala is related to the regulation of nitrogen metabolism and production of secondary metabolites. The biochemistry of plant metabolism against the excess of Cu and Fe TE manifested similarly, but with some specifics regarding the chemical nature of each metal. There was a reduction in the content of ureides and proteins and an increase in amino acids in the roots in relation to the increase in Cu and Fe concentrations. There was low accumulation of proline in the roots in treatments 400 and 500 mg/dm3 compared to the control for both TE. On the other hand, the total phenolic compounds in the roots increased. Our results indicate that the increased synthesis of amino acids and the accumulation of phenolic compounds is involved in the tolerance of L. leucocephala to Cu and Fe.

Transcriptome analysis of changes in M. aeruginosa growth and microcystin production under low concentrations of ethinyl estradiol

Ethinyl estradiol (EE2) is a synthetic environmental estrogen with considerable estrogenic activity. It has been found to consequently pose a significant threat to the aquatic ecosystem. Harmful algal blooms are a major aquatic ecological issue. However, the relationship between EE2 and cyanobacterial bloom is mainly unknown. In this study, the physiological and molecular responses of Microcystis aeruginosa to EE2 exposure were investigated. A low level of EE2 (0.02 μg/L) significantly enhanced the growth of algal cells (P < 0.05), whereas higher concentrations of EE2 (0.2–200 μg/L) inhibited it. EE2 at doses ranging from 0.02 to 200 μg/L promoted the production of microcystins (MCs), with genes mcyABD playing a key role in the regulation of MC synthesis. The alterations of chlorophyll–a, carotenoid, and phycocyanin contents caused by EE2 showed the same trend as cell growth. At the molecular level, 200 μg/L EE2 significantly down–regulated genes in photosynthetic pigment synthesis, light harvesting, electron transfer, NADPH, and ATP generation. High concentrations of EE2 caused oxidative damage to algal cells on the 4th d. After 12d exposure, although there was no significant change in superoxide dismutase (SOD) content and no damage observed in membrane lipids, genes related to SOD and glutathione were changed. In addition, due to the down–regulation of pckA, PK, gltA, nrtA, pstS, etc., carbon fixation, glycolysis, TCA cycle, nitrogen and phosphorus metabolism were hindered by EE2 (200 μg/L). Gene fabG in fatty acid biosynthesis was significantly up–regulated, promoting energy storage in cells. These findings provide important clues to elucidate the effects and mechanisms of cyanobacterial blooms triggered by EE2 and help to effectively prevent and control cyanobacterial blooms.

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.

GCN4 Enhances the Transcriptional Regulation of AreA by Interacting with SKO1 To Mediate Nitrogen Utilization in Ganoderma lucidum.

Fungi utilize a wide range of nitrogen to adapt their metabolism. The transcription factor GCN4 has a pivotal role in nitrogen metabolism. However, the mechanism by which GCN4 regulates nitrogen utilization in Ganoderma lucidum is not well understood. In this study, we found that GCN4 physically interacts with SKO1, a bZIP (basic leucine zipper) transcription factor. GCN4 cooperated with SKO1 to positively regulate nitrogen utilization, especially for the expression of areA. Electrophoretic mobility shift assays (EMSA) indicate that GCN4 directly binds to the areA promoter region. Further affinity analysis through biolayer interferometry (BLI) experiments and surface plasmon resonance (SPR) confirmed that GCN4 specifically binds to the promoter region of areA with a strong binding affinity to activate the transcription of areA. In contrast, SKO1 showed no specified binding effect on the areA promoter. However, SKO1 activates the expression of the areA by forming a complex with GCN4, which exhibits a 14.2-fold-higher affinity than GCN4 alone. Furthermore, the presence of SKO1 promotes the stability of GCN4 protein. Accordingly, our study found that the transcription factor SKO1 enhances the transcriptional activity of GCN4 on its target gene areA by interacting with GCN4. Our study illustrates a specific regulatory mechanism for the involvement of GCN4 and SKO1 in nitrogen utilization, which provides innovative insight into the regulation of nitrogen utilization in fungi. IMPORTANCE Nitrogen is an essential nutrient for cell growth and proliferation. Limitations of nitrogen availability in organisms elicit a series of rapid transcriptional reprogramming mechanisms, which involve the participation of many transcription factors. However, the specific mechanism of coordination between different transcription factors regulating nitrogen metabolism has not been explored. Our study revealed that GCN4 interacts with SKO1 and that they are both involved in regulating nitrogen utilization by affecting the transcription level of areA. We also found that GCN4 activates transcription by directly binding to the promoter recognition region of areA. SKO1 facilitates the transcription of areA by GCN4 by forming a more stable complex with GCN4. Our study deepens our understanding of the regulatory network of nitrogen metabolism and demonstrates a further level of regulation for transcription factors.

Qin-Qiao-Xiao-Du formula alleviate influenza virus infectious pneumonia through regulation gut microbiota and metabolomics

Qin-Qiao-Xiao-Du (QQXD), a traditional Chinese medicine (TCM) formula, has been used in the clinical treatment of influenza virus pneumonia. However, the effects and mechanisms of QQXD on influenza virus pneumonia remain unknown. Therefore, this study explores the mechanisms of QQXD in the treatment of influenza virus pneumonia from the point of view of intestinal flora and metabolism. The results showed that QQXD was able to reduce mortality, weight loss, lung viral load, lung index, and lung injury in influenza virus mice. A cytokine array found that the QQXD attenuated the expression of serum IL-1α, IL-4, IL-12(P70), and TNF-α. Subsequently, 16s rRNA gene sequencing showed that QQXD could increase the relative abundances of Gemmiger, Anaerofustis, Adlercreutzia, and Streptococcus and decrease those of Dehalobacteriu, Burkholderia, Prevotella, Butyrimimonas, Delftia, and others. Meanwhile, targeted metabolic profiling analysis showed that QQXD could regulate nitrogen metabolism, phenylalanine metabolism, valine, leucine, and isoleucine biosynthesis. Correlation analysis demonstrated that the regulatory effect of QQXD on the cyanoamino acid metabolism pathway was associated with changes in the abundance of Parabacteroides, Pediococcus, and Clostridium in influenza mice. In conclusion, our study revealed that QQXD can inhibit influenza virus replication, suppress cytokine storms, and protect mice from influenza virus infection pneumonia. The mechanisms are likely to be related to improved gut microbiota dysbiosis, increased intestinal carbohydrate metabolism, and up-regulated cyanoamino acid metabolism pathways.

Exogenous calcium promotes growth of adzuki bean (Vigna angularis Willd.) seedlings under nitrogen limitation through the regulation of nitrogen metabolism

Plants exhibit lower nitrogen use efficiency (NUE) under N-limitation conditions. Although the function of calcium (Ca) has been widely studied in plants, it remains to be explored whether regulation of nitrate uptake and reduction is needed. A hydroponics experiment on adzuki beans (Vigna angularis Willd.) was used as a test material to determine the interactions between Ca and three levels of nitrogen supply. The height of the plant, the leaf area per plant, the biomass of the plant, the morphology of the roots, the hydraulic conductivity of the roots, the level of gas exchange, and the level of N metabolism of the adzuki beans were evaluated. Furthermore, RT-qPCR was conducted to explore the expression of genes related to nitrate transporter responses to Ca under N-limitation stress conditions. The rate of accumulation of N in plant tissue increased with the application of Ca. However, plant biomass, photosynthetic parameters, and root activity peaked for Ca2+ supply under N-marginal conditions. Further investigation revealed that the activities of nitrate reductase and glutamine synthetase were relatively high. The transcription of the nitrate transporter (VaNRT1.1; VaNRT2.5) was up-regulated in the roots of the Ca-treated plants. Both N-marginal conditions and N deficiency inhibit N absorption and utilization. The favorable effects of Ca on seedling growth and N metabolism under N-marginal conditions were more significant than those under N-deficiency conditions. The supply of Ca2+ is optimal, as it increases NUE by enhancing photosynthesis, N-metabolizing enzyme activities, and NO3 uptake and transport under N-marginal conditions.

Deletion of NGG1 in a recombinant Saccharomyces cerevisiae improved xylose utilization and affected transcription of genes related to amino acid metabolism.

Production of biofuels and biochemicals from xylose using yeast cell factory is of great interest for lignocellulosic biorefinery. Our previous studies revealed that a natural yeast isolate Saccharomyces cerevisiae YB-2625 has superior xylose-fermenting ability. Through integrative omics analysis, NGG1, which encodes a transcription regulator as well as a subunit of chromatin modifying histone acetyltransferase complexes was revealed to regulate xylose metabolism. Deletion of NGG1 in S. cerevisiae YRH396h, which is the haploid version of the recombinant yeast using S. cerevisiae YB-2625 as the host strain, improved xylose consumption by 28.6%. Comparative transcriptome analysis revealed that NGG1 deletion down-regulated genes related to mitochondrial function, TCA cycle, ATP biosynthesis, respiration, as well as NADH generation. In addition, the NGG1 deletion mutant also showed transcriptional changes in amino acid biosynthesis genes. Further analysis of intracellular amino acid content confirmed the effect of NGG1 on amino acid accumulation during xylose utilization. Our results indicated that NGG1 is one of the core nodes for coordinated regulation of carbon and nitrogen metabolism in the recombinant S. cerevisiae. This work reveals novel function of Ngg1p in yeast metabolism and provides basis for developing robust yeast strains to produce ethanol and biochemicals using lignocellulosic biomass.

Biosynthesized hematite nanoparticles mitigate drought stress by regulating nitrogen metabolism and biological nitrogen fixation in Trigonella foenum-graecum

The widespread and increasing use of biosynthetic hematite nanoparticles (HNPs) has led to their application in agriculture. Drought poses a significant threat to crop development and production in agricultural regions around the world. Two varieties of Trigonella foenum-graecum, Afg 1 (drought tolerant) and Afg 3 (drought sensitive) were treated with 20 and 50 mgL−1 HNPs concentrations tested under 5 and 10 days drought stress with control. Both Afg 1 and Afg 3 varieties responded most efficiently at a concentration of 50 mgL−1 HNPs in mitigating drought stress. HNPs reduced proline, nitrate, and ammonia concentrations by 22.4%, 25%, and 37% respectively. They are involved in nitrogen regulation and accumulate in dose-dependent manner during drought stress, causing toxicity. HNPs were found to be important in positively regulating the activity of nitrogen metabolizing enzymes (p ≤ 0.05). Nitrate reductase, glutamine synthetase, and glutamate synthase are increased by 41%, 80%, and 63% respectively. This study suggests that HNPs enhance the ability of Trigonella foenum-graecum to maintain high nitrogen metabolism and biological nitrogen fixation. nifH gene expression increased by 21.7% under drought stress by alleviating the Fe micronutrient deficiency in plants caused by dry soil alkalinity.

The evolution and role of the periplasmic asparaginase Asp3 in yeast.

ABSTRACTThe study of nitrogen assimilation in yeast is of interest from genetic, evolutionary, and biotechnological perspectives. Over the course of evolution, yeasts have developed sophisticated control mechanisms to regulate nitrogen metabolism, with domesticated lineages sometimes displaying particular specialisation. The focus of this study was on assimilation of asparagine, which is a significant nutritional source for some alcoholic fermentations. We were particularly interested in ASP3, which encodes a periplasmic asparaginase and that was proposed to have been acquired relatively recently in S. cerevisiae by horizontal gene transfer. We examined 1680 S. cerevisiae genome assemblies to evaluate the distribution and evolutionary trajectory of ASP3. Our findings suggest an alternative hypothesis that ASP3 is an ancient Saccharomyces gene that has generally been lost over the course of evolution but has been retained in certain fermentative environments. As asparagine is the major nitrogen source in apple juice, we explored whether the presence of ASP3 would confer a growth advantage. Interestingly, we found that although ASP3 enhances growth when asparagine is the sole nitrogen source, the same effect is not seen in apple juice. These data indicate that growth in pure culture may not reflect the original selective environment for ASP3+ strains and highlight the role that complex regulation may play in optimising nitrogen assimilation in yeasts.

Calcium sustains K+-Na+ homeostasis through regulating nitrogen metabolism in hydrogen sulfide-dependent manner in tomato roots under salinity

The work presented here exhibits the function of calcium (Ca) and hydrogen sulfide (H2S) in the tolerance of tomato seedlings against NaCl stress. The NaCl (100 mM)-stressed seedlings supplemented with 20 mM Ca, showed a substantial increase in K+/Na+ ratio which however, exhibited a significant decrease when they did not receive Ca. Using plasma membrane inhibitors, it was observed that Ca maintained a higher K+/Na+ ratio by altering K+ and Na+ transport via regulation of Na+-H+ antiport system. However, a reverse trend was noticed when 1 mM hypotaurine (HT; H2S scavenger) was applied along with Ca which validates the role of H2S in Ca-mediated regulation of K+-Na+ homeostasis. Interactive effect of Ca and H2S also improved the activity of the enzymes involved in nitrogen (N) metabolism. Impact of enhanced activity of the enzymes of N metabolism was also reflected in increased inorganic N forms viz. nitrate, nitrite, and ammonium. The stressed seedlings supplied with Ca exhibited higher activity of antioxidant enzymes (superoxide dismutase, catalase, and peroxidase) which maintained optimum level of reactive oxygen species (ROS) which was higher in the stressed seedlings that did not receive Ca. A reduction in ROS by Ca led to reduced lipid peroxidation and electrolyte leakage than the NaCl-stressed seedlings. On the other hand, application of HT exhibited a reverse trend in these parameters even in presence of Ca. It signifies H2S-dependent functioning of Ca. However, a non-significant effect of HT on root Ca content suggests upstream functioning of Ca to H2S. Therefore, it can be postulated that Ca, through regulating N metabolism, protected the seedlings against NaCl-induced constrains in H2S-dependent manner, and Ca functions upstream of H2S in the regulation of plant adaptive mechanisms to salinity.

Cat8 Response to Nutritional Changes and Interaction With Ehrlich Pathway Related Factors.

Cat8 is an important transcription factor regulating the utilization of non-fermentative carbon sources in Saccharomyces cerevisiae. However, our previous studies found that Cat8 may play a critical role in nitrogen metabolism, but the regulatory mechanism has not been elucidated. In this study, the nuclear localization and analysis of regulatory activity showed that the Cat8 function relies on Snf1 kinase. In the fermentation with glucose or glycerol as carbon sources under phenylalanine (Phe) induction, by comparing the changes of cellular gene expression and Cat8 target gene binding profiles after Cat8 overexpression, enhanced transcription was shown among key genes involved in the Ehrlich pathway (e.g., ARO9, ARO10, and ADH2) and its upstream and downstream related factors (e.g., GAP1, AGP1, GAT1, PDR12, and ESPB6), indicating that Cat8 participated in the regulation of nitrogen metabolism. Moreover, highly active Cat8 interacts with transcriptional activator Aro80 and GATA activator Gat1 coordinately to regulate the transcription of ARO10. Altogether, our results showed that Cat8 may act as a global transcription factor in response to nutritional changes, regulating both carbon and nitrogen utilization. This provides a new insight for us to explore the regulation of cell nutrient metabolism networks in yeast.

Mass Spectrometric Studies of Valine Molecules by Electron Shock in the Gas Phase

Relevance. Valine is one of the eight amino acids not synthesised by the human body, necessary for the synthesis and growth of body tissues, muscle coordination; regulation of nervous processes, nitrogen metabolism, and stabilisation of the hormonal background. Since α-amino acids contain an asymmetric carbon atom, they can exist as optical isomers (mirror antipodes) that play an essential role in protein biosynthesis. The structure of matter and the physical processes that occur in it are studied using the method of mass spectrometry and spectral analysis. This indicates the relevance of the problem that was studied in this paper. Purpose. Mass spectrometric studies of the formation of ionic products of single and dissociative ionisation of the valine molecule (C5 H11NO2) with electrons according to the method of beams intersecting within the energy range of bombarding electrons 6-70 eV. To consider the mechanisms of formation of the most intense ion fragments during dissociative ionisation by electron shock. Methods. The experiment was conducted on an installation with a monopole mass spectrometer of the MX-7304A type, which belongs to the class of dynamic mass analysers with electron shock ionisation in the range of mass numbers 0-120 Da. The mass spectra of molecules were investigated at different temperatures of the source of molecules in the range of 300-600 K. Results. The obtained results are compared with the mass spectra of the D-, L-, and DL-enantiomeric forms of the valine molecule with data from the NIST and SDBS databases. The features of the processes of formation of ion fragments of valine molecules by electronic shock are analysed in detail, and the dynamics of the yield of ion fragments in the range of evaporation temperatures of the initial substance of 300-440 K is also studied. The total relative ionisation cross-section of the molecule under study was measured according to mass spectrometric method with an ionising electron energy of 5-60 eV. Based on the results of experimental studies, a threshold section of the dependence of the total relative cross-section of valine ionisation is determined and given in this paper. Conclusions. A detailed analysis of the processes of formation of fragment ions in the mass spectra allows demonstrating the influence of the structural forms of valine enantiomers on the redistribution of relative intensities of product ions

Transcription Regulation of Cell Cycle Regulatory Genes Mediated by NtrX to Affect Sinorhizobium meliloti Cell Division

The cell division of the alfalfa symbiont, Sinorhizobium meliloti, is dictated by a cell cycle regulatory pathway containing the key transcription factors CtrA, GcrA, and DnaA. In this study, we found that NtrX, one of the regulators of nitrogen metabolism, can directly regulate the expression of ctrA, gcrA, and dnaA from the cell cycle pathway. Three sets of S. meliloti ntrX mutants showed similar cell division defects, such as slow growth, abnormal morphology of some cells, and delayed DNA synthesis. Transcription of ctrA and gcrA was upregulated, whereas the transcription of dnaA and ftsZ1 was downregulated in the insertion mutant and the strain of Sm1021 expressing ntrXD53E. Correspondingly, the inducible transcription of ntrX activates the expression of dnaA and ftsZ1, but represses ctrA and gcrA in the depletion strain. The expression levels of CtrA and GcrA were confirmed by Western blotting. The transcription regulation of these genes requires phosphorylation of the conserved 53rd aspartate in the NtrX protein that binds directly to the promoter regions of ctrA, gcrA, dnaA, and ftsZ1 by recognizing the characteristic sequence CAAN2-5TTG. Our findings suggest that NtrX affects S. meliloti cell division by regulating the transcription of the key cell cycle regulatory genes.

Analysis of Whole-Transcriptome RNA-Seq Data Reveals the Involvement of Alternative Splicing in the Drought Response of Glycyrrhiza uralensis.

Alternative splicing (AS) is a post-transcriptional regulatory mechanism that increases protein diversity. There is growing evidence that AS plays an important role in regulating plant stress responses. However, the mechanism by which AS coordinates with transcriptional regulation to regulate the drought response in Glycyrrhiza uralensis remains unclear. In this study, we performed a genome-wide analysis of AS events in G. uralensis at different time points under drought stress using a high-throughput RNA sequencing approach. We detected 2,479 and 2,764 AS events in the aerial parts (AP) and underground parts (UP), respectively, of drought-stressed G. uralensis. Of these, last exon AS and exon skipping were the main types of AS. Overall, 2,653 genes undergoing significant AS regulation were identified from the AP and UP of G. uralensis exposed to drought for 2, 6, 12, and 24 h. Gene Ontology analyses indicated that AS plays an important role in the regulation of nitrogen and protein metabolism in the drought response of G. uralensis. Notably, the spliceosomal pathway and basal transcription factor pathway were significantly enriched with differentially spliced genes under drought stress. Genes related to splicing regulators in the AP and UP of G. uralensis responded to drought stress and underwent AS under drought conditions. In summary, our data suggest that drought-responsive AS directly and indirectly regulates the drought response of G. uralensis. Further in-depth studies on the functions and mechanisms of AS during abiotic stresses will provide new strategies for improving plant stress resistance.

Chromatin Regulators Ahc1p and Eaf3p Positively Influence Nitrogen Metabolism in Saccharomyces cerevisiae.

There is a complex regulatory network of nitrogen metabolism in Saccharomyces cerevisiae, and many details of this regulatory network have not been revealed. This study explored the global regulation of nitrogen metabolism in S. cerevisiae from an epigenetic perspective. Comparative transcriptome analysis of S. cerevisiae S288C treated with 30 nitrogen sources identified nine chromatin regulators (CRs) that responded significantly to different nitrogen sources. Functional analysis showed that among the CRs identified, Ahc1p and Eaf3p promoted the utilization of non-preferred nitrogen sources through global regulation of nitrogen metabolism. Ahc1p regulated nitrogen metabolism through amino acid transport, nitrogen catabolism repression (NCR), and the Ssy1p-Ptr3p-Ssy5p signaling sensor system. Eaf3p regulated nitrogen metabolism via amino acid transport and NCR. The regulatory mechanisms of the effects of Ahc1p and Eaf3p on nitrogen metabolism depended on the function of their histone acetyltransferase complex ADA and NuA4. These epigenetic findings provided new insights for a deeper understanding of the nitrogen metabolism regulatory network in S. cerevisiae.

Nitrogen metabolism in different configuration design constructed wetlands under exposure of graphene oxide

With occurrence of emerging contaminants in wastewater, nitrogen metabolism in constructed wetlands (CWs) should be re-evaluated at different environmental variables to withstand adversely exogenous contaminants. This study investigated functional gene, key enzyme, and microbe regulating nitrogen metabolism in three different configuration design CWs (planted W1 and W3 with full and low water level, unplanted W2 with full water level) when exposed to graphene oxide, one of most popular carbon-based nanomaterials. It was found that nitrogen removal was achieved by classical nitrification and denitrification, which was limited by nitrification in W2 and denitrification in planted CWs, presenting optimal balance between nitrification and denitrification in W1. Compared to W2, ammonium removals increased by 48% and 107% in W1 and W3, conversely total nitrogen removals decreased by 6% and 14% due to nitrate accumulation. Ammonia monooxygenase was the most sensitive to variables among four key enzymes. Bacteria were more abundant than archaea in CWs, both of functional genes and nitrifying populations were obviously affected by plant and water level in later period. The dominant denitrification populations were class Gammaproteobacteria followed by Alphaproteobacteria with the highest abundance observed in W2. Their species richness at genus level was higher on day 30 in W2 than planted CWs, but similar on day 120 in three CWs. The cooccurrence network and associated heatmap evidenced complex interaction among key enzymes, functional genes, and functional genera for nitrogen removal. KEGG annotations revealed specific differences in six core pathways of nitrogen metabolism in three CWs.