GDH Activity Research Articles

Alterations in nitrogen metabolism caused by heavy metals in the acid-tolerant microalga Coccomyxa onubensis

The microalga Coccomyxa onubensis is an extremophile microorganism with a unique ecosystem (Río Tinto, Huelva, Spain) that contains high amounts of contaminants, including heavy metals, sulphates, and nitrates, in acidic environments (pH 2.5). The current work presents an evaluation of the capacity of Coccomyxa onubensis to assimilate different nitrogen sources under Cu2+, Cd2+, AsO33−, AsO43− and Hg2+ stress, and the metabolic implications of these stressors. The results showed that ammonium consumption was less affected than nitrate consumption when microalgae were cultivated with heavy metals (except cadmium). The activities of enzymes involved in nitrogen metabolism, such as nitrite reductase (NiR; EC:1.7.7.1), glutamine synthetase (GS; EC:6.3.2.1) and glutamate dehydrogenase (GDH; EC:1.4.1.2) were characterised to determine the Michaelis-Menten constant (Km) and optimal temperature and pH values, being 45, 40 and 60 °C and pH values of 7.5, 6.0 and 9.0 for NiR, GS, and GDH, respectively. The effects of different heavy metals on these enzymes were assessed at multiple levels, and the results showed that the enzymatic activity of NiR was downregulated, specially under copper stress, maintaining 23 % of control NiR activity at 2 mM Cu2+. The enzymatic activity of GS was upregulated at low concentrations under cadmium and mercury stress (115–120 % of control cultures GS activity at 25 μM Cd2+ and 50 nM Hg2+, respectively) and downregulated at high concentrations of these elements. GDH activity was upregulated in the presence of Cu2+, Cd2+, and Hg2+, with increases up to 192, 155 and 154 % at 1 mM Cu2+, 300 μM Cd2+, and 250 nM Hg2+, respectively. These results provide a better explanation of the effects of heavy metal stress on N metabolism in Coccomyxa onubensis, which may be used as a model eukaryotic organism of the Tinto River acidophilic ecosystem.

A Versatile Method to Create Antibody/Split‐Enzyme Complexes and Its Application to a Rapid, Homogeneous, and Universal Electrochemical Immunosensing System

AbstractIn this study, a versatile method is developed to create an antibody/split‐enzyme complex for use in electrochemical immunosensors. SpyCatchers are genetically fused at each terminus of flavin adenine dinucleotide‐dependent glucose dehydrogenase (FAD‐GDH) and a protease recognition sequence is inserted into a loop region to prepare a soluble protein and the split GDH. SpyTag‐fused single‐chain variable fragments (scFvs) are employed, which leads to the simple preparation of antibody‐split enzyme complexes (split AECs). The addition of pentameric C‐reactive protein (CRP), as a representative multimeric protein, restored the GDH activity of the split AECs, as the two split AEC fragments formed active GDH when in close proximity. CRP in human serum is electrochemically detected within 5 min without any washing steps with a clinically relevant CRP detection range (0.03–1 mg dL−1). The detection system is expanded to detect hemoglobin, SARS‐CoV‐2 spike protein, and inactivated SARS‐CoV‐2 by exchanging the scFvs. These results show that the convenient preparation method of the split GDH and the split AEC by the insertion of the protease recognition sequence and the use of SpyCatcher/SpyTag can realize a rapid, homogeneous, and universal electrochemical detection system that can be integrated into a commercially available electrochemical glucose sensor.

Natural variation in the adjustment of primary metabolism determines ammonium tolerance in the model grass Brachypodium distachyon.

Nitrogen (N) fertilization is essential to maximize crop production. However, around half of the applied N is lost to the environment causing water and air pollution and contributing to climate change. Understanding the natural genetic and metabolic basis underlying plants N use efficiency is of great interest to reach an agriculture with less N demand and thus, more sustainable. The study of ammonium (NH4+) nutrition is of particular interest, because it mitigates N losses due to nitrate (NO3-) leaching or denitrification. In this work, we studied Brachypodium distachyon, the model plant for C3 grasses, grown with NH4+ or NO3- supply. We performed gene expression analysis in the root of the B. distachyon reference accession Bd21 and examined the phenotypic variation across 52 natural accessions through analysing plant growth and a panel of 22 metabolic traits in leaf and root. We found that the adjustment of primary metabolism to ammonium nutrition is essential for the natural variation of NH4+ tolerance, notably involving NH4+ assimilation and PEPC activity. Additionally, genome-wide association studies (GWAS) indicated several loci associated with B. distachyon growth and metabolic adaptation to NH4+ nutrition. For instance, we found that the GDH2 gene was associated with the induction of root GDH activity under NH4+ nutrition and that two genes encoding malic enzyme were associated with leaf PEPC activity. Altogether, our work underlines the value of natural variation and the key role of primary metabolism to improve NH4+ tolerance.

Redefining the Role of Ornithine Aspartate and Vitamin E in Metabolic-Dysfunction-Associated Steatotic Liver Disease through Its Biochemical Properties.

It is known that the inflammation process leading to oxidative stress and thyroid hormone metabolism dysfunction is highly altered in metabolic dysfunction associated with steatotic liver disease (MASLD). This study aims to address the effect of ornithine aspartate (LOLA) and vitamin E (VitE) in improving these processes. Adult Sprague-Dawley rats were assigned to five groups and treated for 28 weeks: controls (n = 10) received a standard diet (for 28 weeks) plus gavage with distilled water (DW) from weeks 16 to 28. MASLD groups received a high-fat and choline-deficient diet for 28 weeks (MASLD group) and daily gavage with 200 mg/kg/day of LOLA, or twice a week with 150 mg of VitE from weeks 16-28. LOLA diminished collagen deposition (p = 0.006). The same treatment diminished carbonyl, TBARS, and sulfhydryl levels and GPx activity (p < 0.001). Type 3 deiodinase increased in the MASLD group, downregulating T3-controlled genes, which was corrected in the presence of LOLA. LOLA also promoted a near-normalization of complex II, SDH, and GDH activities (p < 0.001) and improved reticulum stress, with a reduction in GRP78 and HSPA9/GRP75 protein levels (p < 0.05). The enhanced energy production and metabolism of thyroid hormones, probably because of GSH replenishment provided by the L-glutamate portion of LOLA, opens a new therapeutic approach for MASLD.

The Effects of Tillage and the Combined Application of Organic and Inorganic Fertilizers on the Antioxidant Enzyme Activity and Yield of Maize Leaves

The aim of this study was to explore the characteristics of the combined application of organic fertilizer and inorganic fertilizer using different tillage methods to delay the senescence of maize leaves. The yield and activities of GDH, CAT, APX, GR, and GSH enzymes in maize leaves were measured at different growth stages by using two tillage methods, three organic and inorganic combined applications (P1, P2, and P3), and four control treatments. (1) During the growth period, the R + S and R treatments were P1 treatments, with the highest enzyme activities noted for GDH, CAT, APX, GR, and GSH, which were 36.79–103.22% higher than those of CK. (2) The average yield of all R + S treatments was higher than that of R treatments, and the average yield of P1 treatment was the highest under R + S, which was 13,663.79 kg hm−2, which was 6.39%, 7.90%, and 14.67% higher than that of P2, P3, and CK, respectively, which was lower than that of R. The yield of P1 treatment was 2.53% higher. (3) There was a significant positive correlation between APX activity, CAT activity, GR activity, GDH activity, GSH activity, grain number per ear, ear length, and 100-grain weight of maize leaves at the grain filling stage, and a significant negative correlation between bald tip length and yield. The treatment details had the strongest enzyme activity and the highest yield when using the rotary tillage + subsoiling (R + S) P1 method, which was the most suitable tillage method and the best fertilizer ratio combination, which could be demonstrated and popularized in a large area in the dry farming area of spring maize in Shanxi Province.

Dissolved oxygen and ammonia affect ammonia production via GDH/AMPK signaling pathway and alter flesh quality in Chinese perch (Siniperca chuatsi).

The dissolved oxygen (DO) and ammonia are crucial to the growth of Chinese perch (Siniperca chuatsi). Information on the effects of DO and total ammonia nitrogen (TAN) in regulating ammonia nitrogen excretion and flesh quality in Chinese perch is scanty. This study aimed to evaluate the effects of dissolved DO at oxygen levels of 3 mg/L and 9 mg/L, as well as the TAN concentrations of 0.3 mg/L and 0.9 mg/L on ammonia excretion and flesh quality. Results showed that the ammonia contents in plasma, muscle, and liver of the 9 mg/L DO group were significantly higher than those of the 3 mg/L DO group (P < 0.05). However, the expression of AMPK-related signaling pathway genes (gdh, lkb1, and ampd) and flesh quality indicators (gumminess, chewiness, hardness) in the 9 mg/L DO group were significantly lower than those in the 3 mg/L DO group. Under long-term exposure to 0.9 mg/L TAN, the ammonia contents in plasma and gill filaments, as well as muscle flesh quality (resilience, gumminess, chewiness, cohesiveness), were significantly lower than those in the 0.3 mg/L TAN group (P < 0.05). However, the activities of GDH and AMPD enzymes in the 0.9 mg/L TAN group were significantly higher than those in the 0.3 mg/L TAN group. In summary, when fish are exposed to 3 mg/L DO and 0.9 mg/L TAN in the environment for a long time, their amino acids are used for transamination and deamination, resulting in insufficient energy supply for Chinese perch, whereas 9 mg/L DO and 0.9 mg/L TAN caused deterioration of the flesh quality.

Effect of Nutraceutical Factors on Hepatic Intermediary Metabolism in Wistar Rats with Induced Tendinopathy.

Tendinopathy (TP) is a complex clinical syndrome characterized by local inflammation, pain in the affected area, and loss of performance, preceded by tendon injury. The disease develops in three phases: Inflammatory phase, proliferative phase, and remodeling phase. There are currently no proven treatments for early reversal of this type of injury. However, the metabolic pathways of the transition metabolism, which are necessary for the proper functioning of the organism, are known. These metabolic pathways can be modified by a number of external factors, such as nutritional supplements. In this study, the modulatory effect of four dietary supplements, maslinic acid (MA), hydroxytyrosol (HT), glycine, and aspartate (AA), on hepatic intermediary metabolism was observed in Wistar rats with induced tendinopathy at different stages of the disease. Induced tendinopathy in rats produces alterations in the liver intermediary metabolism. Nutraceutical treatments modify the intermediary metabolism in the different phases of tendinopathy, so AA treatment produced a decrease in carbohydrate metabolism. In lipid metabolism, MA and AA caused a decrease in lipogenesis at the tendinopathy and increased fatty acid oxidation. In protein metabolism, MA treatment increased GDH and AST activity; HT decreased ALT activity; and the AA treatment does not cause any alteration. Use of nutritional supplements of diet could help to regulate the intermediary metabolism in the TP.

Substituting green light for partial red light promoted the growth and quality, and regulated the nitrogen metabolism of Medicago sativa grown under red-blue LEDs

Light quality can affect the growth of plants, regulate the accumulation and metabolism of substances, thereby improving the growth and quality of plants. Alfalfa is a vital forage with wide distribution and high value for feeding. However, few studies have investigated the effects of light quality on growth, quality, and nitrogen (N) metabolism regulation of alfalfa under artificial light control environment. In this study, the light intensity was 400 μmol·m−2·s−1 and photoperiods was 16/8 h. Five light quality treatments were used for alfalfa production, red-blue ratio of 3:1, 4:1 and 5:1, respectively, and on the basis of red-blue ratio of 4:1, 5% and 10% red light were replaced by green light were also used (R3B1, R4B1, R5B1, R19B5G1 and R18B5G2). The results showed that although the stomatal conductance and lysine content of the R3B1 treatment were significantly higher than other treatments, but the leaf area, fresh yield, contents of soluble sugar, sucrose, nitrate and ammonium, as well as the enzyme activities of NR, NiR, GOGAT and GS were the lowest among all treatments. The R3B1 treatment resulted in growth inhibition and reduced N metabolism levels. Strikingly, compared with the R4B1 treatment, the synthesis of Chl a+b, Chl a, Chl b, and free amino acid, as well as the accumulation of nitrate and crude protein, and the activity of GS also increased significantly in the two treatments which green light replaced partial of red light. However, the contents of soluble sugar, glutamate and lysine, as well as the activities of GOGAT and GDH, were significantly reduced in the R19B5G1 and R18B5G2 treatments. The carotenoids and sucrose contents of R19B5G1 treatment were significantly higher than R4B1 treatment, and the fresh weight and NR activity of R18B5G2 were significantly higher than R4B1 treatment. In addition, the content of starch and soluble protein and AK activity were not affected by light quality. In general, green light instead of partial red light increased the content of photosynthetic pigment and promoted the growth of alfalfa. Although the levels of glutamate, cysteine and lysine were reduced, it was beneficial to the accumulation of nitrate, free amino acid and crude protein, maintained the stability of N metabolism and improved the feeding value. These findings provide valuable insight into the effects of light quality regulation on value of feeding and N metabolism of alfalfa.

The NIN-LIKE PROTINE1 (CsNLP1) transcription factor is involved in modulating the nitrate response in cucumber seedlings

Cucumber is one of the important vegetables cultivated in facilities in China. Due to the deterioration of soil quality in facilities and the low nitrogen use efficiency of existing varieties, it is common to pursue high yield through excessive application of nitrogen fertilizer. Improving crop nitrogen use efficiency is one of the important means to reduce nitrogen fertilizer input. Previous studies have shown that NLP (NIN-like protein) transcription factor family play an important role as transcription factors in regulating plant physiological responses to external nitrogen changes. However, there are few reports on NLPs in cucumber. Here we report that CsNLP1 is a potential candidate to improve cucumber nitrogen use ability. CsNLP1 silencing lines by virus-induced gene silencing (VIGS) showed a significant decrease in the plant biomass, chlorophyll content, nitrogen uptake, total nitrogen content, activities of NR, NIR, GDH, GOGAT and GS, and expression levels of genes involved in nitrogen assimilation and signalling under different nitrogen concentrations. In addition, transfer of CsNLP1 into Arabidopsis nlp7-1 mutant found that the phenotype, nitrate (NO3-) content, NR activity, NIR activity, GDH activity, GOGAT activity and GS activity in transgenic lines could be restored to the wild type level after NO3- treatment. Further study found that CsNLP1 could regulate the expression of NO3- responsive genes such as AtNIR1, AtNIA2, AtNRT1.1 and AtGS2 in plants. Overall, our data indicated that CsNLP1 can affect plant growth, and regulate nitrate assimilation by regulating nitrogen-related genes.

The Relationship between Treatment Response in Patients with Acute Psychotic Episodes of Schizophrenia and the Activity of Their Platelet Enzymes

Background: imbalance in the energy processes, such as oxidative phosphorylation intensity, and antioxidant systems, and impaired glutamate metabolism in the brain are important biochemical pathogenetic links in the development of schizophrenia. Changes in the activity of a number of platelet enzymes involved in these biochemical pathways were found in patients with schizophrenia.Aim: to assess the relationship between the treatment effciency of acute psychotic episode in patients with paranoid schizophrenia and the activity levels of platelet enzymes involved in energy metabolism (cytochrome c-oxidase, COX), in the antioxidant glutathione system (glutathione reductase, GR), and glutamate metabolism (glutamate dehydrogenase, GDH).Patients and methods: we examined hospitalized adult patients with a diagnosis of paranoid schizophrenia, episodic course, exacerbation of the disease (F20.01), receiving antipsychotic therapy, and healthy volunteers as a control group for assessment of control ranges for enzymatic activities. Psychometric assessment was carried out using the Positive and Negative Syndrome Scale (PANSS). Treatment response was assessed as good with a reduction in PANSS scores of 20% or more. The evaluation was performed on the first days after hospitalization and immediately before discharge.Results: 113 subjects were recruited. 50 healthy volunteers formed the control group and 63 patients, including 60 men and 3 women made up the study group. All indicators were obtained in all control group members and in 42 patients. In the group with a good treatment response, the baseline COX, GDH and GR activities proved to be significantly more often met within the control range, and the GR activity in half of the patients increased after the treatment course. In the group with insuffcient response, half of the patients had an increase in GDH activity after treatment. Of all the biochemical indices, the parameter most associated with therapeutic response was GDH activity.Conclusion: GDH activity can be considered as a possible candidate for predicting the therapeutic response to antipsychotic therapy.

Mitigation of Salt Stress in Reaumuria soongarica Seedlings by Exogenous Ca2+ and NO Compound Treatment

Soil salinization is a common abiotic stress that severely limits the growth of Reaumuria soongarica and reduces its application value. To better understand the response of R. soongarica to salt stress and the physiological mechanisms of exogenous Ca2+ and NO compound treatment in alleviating salt stress, the growth parameters, antioxidant system, carbohydrate metabolism and nitrogen compound metabolism were compared on Days 0, 1, 3, 6, 9, 15 and 30. The results showed that salt stress could significantly reduce the plant height, root length, fresh and dry weights of aboveground and underground parts, as well as the relative water content, severely inhibiting the growth of R. soongarica seedlings. After Ca2+ and NO compound treatment, these growth parameters were significantly improved, and the harm caused by stress in R. soongarica was alleviated. Regarding the antioxidant system, the Ca2+ and NO compound treatment could significantly increase the activities of SOD, CAT, APX and GR, as well as the contents of ASA and GSH, which indicated that exogenous Ca2+ and NO could eliminate the accumulated active oxygen by increasing the activities of oxidoreductases and the content of nonenzymatic antioxidant substances, thereby improving the salt tolerance of R. soongarica. Regarding carbon metabolism, after Ca2+ and NO compound treatment, the soluble sugar and sucrose contents, as well as the activities of sucrose phosphate synthase and sucrose synthase, were significantly increased, which indicated that Ca2+ and NO compound treatment could maintain higher soluble sugar and sucrose contents in R. soongarica and reduce osmotic stress caused by salt treatment. Regarding nitrogen metabolism, the Ca2+ and NO compound treatment reduced the harm of salt stress by regulating the nitrogen compound contents and nitrogen compound-related enzyme activities, including increases in the NO3− content and NR, NiR, GS, GOGAT and GDH activities and a reduction in the NO2− content. The results of this study indicate that the inhibition of the growth and development of R. soongarica by salt stress can be alleviated by regulating the antioxidant system, carbohydrate metabolism and nitrogen compound metabolism, which provides a theoretical basis for Ca2+ and NO compound treatment to improve plant salt tolerance.

Effects of Exogenously Applied Copper in Tomato Plants’ Oxidative and Nitrogen Metabolisms under Organic Farming Conditions

Currently, copper is approved as an active substance among plant protection products and is considered effective against more than 50 different diseases in different crops, conventional and organic. Tomato has been cultivated for centuries, but many fungal diseases still affect it, making it necessary to control them through antifungal agents, such as copper, making it the primary form of fungal control in organic farming systems (OFS). The objective of this work was to determine whether exogenous copper applications can affect AOX mechanisms and nitrogen use efficiency in tomato plant grown in OFS. For this purpose, plants were sprayed with ‘Bordeaux’ mixture (SP). In addition, two sets of plants were each treated with 8 mg/L copper in the root substrate (S). Subsequently, one of these groups was also sprayed with a solution of ‘Bordeaux’ mixture (SSP). Leaves and roots were used to determine NR, GS and GDH activities, as well as proline, H2O2 and AsA levels. The data gathered show that even small amounts of copper in the rhizosphere and copper spraying can lead to stress responses in tomato, with increases in total ascorbate of up to 70% and a decrease in GS activity down to 49%, suggesting that excess copper application could be potentially harmful in horticultural production by OFS.

Studies of the silkworm enzyme activity and their correlations with economic variables

The experiment was conducted to analyse the larval performance and economic traits of bivoltine silkworm breeds of silkworm (SK-1, SK-6, SK-22, SK-28, SK-33, CSR4, CSR2, NB4D2, DUN6 and APS4) during spring season. The haemolymph total protein, succinate and gultamate dehydrogenase activities were estimated and their correlation with economic traits were also worked out. The results of the study confirmed that among ten bivoltine silkworm breeds, highest haemolymph SDH activity of 3.47 µmoles/ml/mgprotein/min was recorded in the silkworm breed SK1 and least SDH activity of 1.58 µmoles/ml/mg protein/min was recorded in the breed APS4. The highest peak of succinate dehydrogenase activity of 2.65 µmoles/ml/mg protein/min was observed on 7th day of the 5th instar and lowest peak of succinate dehydrogenase activity of 2.39 µmoles/ml/mg protein/min was observed on 4th day of the 5th instar. GDH activity of 0.46 µmoles/ml/mg protein/min was recorded highest in the silkworm breed SK1 and lowest of 0.15 µmoles/ml/mg protein/min was recorded in silkworm breed APS4. The highest peak of haemolymph GDH of 0.36 µmoles/ml/mg protein/min was recorded on 7th day of 5th instar and lowest peak of 0.26 µmoles/ml/mg protein/min was recorded on 4th day of 5th instar. The correlational studies revealed that haemolymph total protein, SDH and GDH were found to be positively corelated with yield by weight and number (cocoon), weight of mature larvae, shell weight ,cocoon weight, shell ratio percent, silk productivity, rate of pupation, fecundity, raw silk percentage and length of filament. Thus, the study revealed that silkworm breeds like SK1, SK6, SK22 and SK28 as productive breeds and hence may be used for future breeding programmes for evolution of new robust silkworm breeds

Ectopic Expression of Sugarcane ScAMT1.1 Has the Potential to Improve Ammonium Assimilation and Grain Yield in Transgenic Rice under Low Nitrogen Stress.

In China, nitrogen (N) fertilizer is excessively used in sugarcane planting areas, while the nitrogen use efficiency (NUE) of sugarcane is relatively low. Mining and identifying the key genes in response to low N stress in sugarcane can provide useful gene elements and a theoretical basis for developing sugarcane varieties with high NUE. In our study, RNA-Seq combined with qRT-PCR analysis revealed that the ScAMT1.1 gene responded positively to low N stress, resulting in the stronger low N tolerance and high NUE ability of sugarcane cultivar ROC22. Then, ScAMT1.1 was cloned from sugarcane. The full-length cDNA of the ScAMT1.1 gene is 1868 bp, containing a 1491 bp open reading frame (ORF), and encoding 496 amino acids. ScAMT1.1 belongs to the AMT superfamily and shares 91.57% homologies with AMT1.1 from Oryza sativa. Furthermore, it was stably overexpressed in rice (O. sativa). Under low N treatment, the plant height and the fresh weight of the ScAMT1.1-overexpressed transgenic rice were 36.48% and 51.55% higher than that of the wild-type, respectively. Both the activity of ammonium assimilation key enzymes GS and GDH, and the expression level of ammonium assimilation key genes, including GS1.1, GS1.2, GDH, Fd-GOGAT, and NADH-GOGAT2 in the transgenic plants, were significantly higher than that of the wild-type. The grain number and grain yield per plant in the transgenic rice were 6.44% and 9.52% higher than that of the wild-type in the pot experiments, respectively. Taken together, the sugarcane ScAMT1.1 gene has the potential to improve ammonium assimilation ability and the yield of transgenic rice under low N fertilizer conditions. This study provided an important functional gene for improving sugarcane varieties with high NUE.

Transcriptomic and enzymatic analysis reveals the roles of glutamate dehydrogenase in Corynebacterium glutamicum

Glutamate dehydrogenase (Gdh), catalyzing the reversible conversion between 2-oxoglutarate and glutamate, plays an important role in the connection of nitrogen and carbon metabolism. Yet little is known about these enzymes in the amino acid-manufacturing Corynebacterium glutamicum. In the present study, we firstly identified the enzymatic characteristics of two Gdhs (GdhA and GdhB). The results showed that both GdhA and GdhB prefers NADPH as a coenzyme and have higher affinity for 2-OG than glutamate. The growth characteristics of gdhAΔ mutant and gdhBΔ mutant, gdhABΔ mutant showed GdhA serves as the main conduit for ammonium assimilation, and GdhB is the main glutamate- metabolizing enzyme in C. glutamicum. The full-genome transcriptomic analysis was used to investigate physiological response of C. glutamicum to the glutamate as nitrogen source, and gdh deletion. The results showed that the nitrogen starvation response was elicited when glutamine served as the sole nitrogen source. gdhAΔBΔ double deletion trigger a partially deregulated nitrogen starvation response, in which genes involved in nitrogen assimilation showed obviously upregulated in a certain extent. On the other hand, the genes of phosphotransferase system (PTS) and glycolysis pathway, most genes in pentose phosphate pathway were significantly upregulated, indicating that gdh deficiency initiated the enhancement of the absorption and metabolism of carbon sources. We believed that our results in this study will give new insights on the molecular mechanism of Gdh activity cross-talks with carbon and nitrogen metabolism, also setting a new background for further flux redistribution applied research of biotechnological interest.

Transcriptomic dissection underlying physiological and anatomical characteristics of poplar wood in response to changes in light intensity and nitrogen availability

To dissect the transcriptomic reprograming underlying the physiological and anatomical characteristics in the wood of Populus species in response to changes in light and nitrogen (N) availability, the saplings of P. alba × P. glandulosa were exposed to either control or high light intensity in combination with one of low, normal and high N levels. High light caused greater CO2 assimilation rate, higher concentrations of soluble sugars and starch, decreased enzymatic activities of NR and GS and increased activities of GOGAT and GDH, higher IAA and lower ABA, JA and SA contents, narrower fibers, and thinner thickness of fiber wall in the wood of poplars under normal N condition. Low N supply led to greater levels of sucrose and starch, lower contents of most amino acids, IAA, ABA and JA, and narrower vessels and fibers, and high N caused the opposite effects in poplar wood exposed to control light. Transcriptomic reprograming occurred underlying the physiological and anatomical changes of P. alba × P. glandulose in acclimation to high light in combination with one of low, normal and high N levels. High light- and low/high N-induced differential expression of the key genes including neutral invertases, ASN1, JAZ1, and TIP1;3, involved in the metabolism of carbohydrate, amino acids and phytohormones, and water transport, corresponded well to changes in physiological and anatomical characteristics in poplar wood in response to high light and altered N availability. These results suggest that physiological and anatomical characteristics are altered in the wood and the underlying transcriptomic reprograming is critical for the wood in acclimation to high light in combination with changing N availability.

Evaluation of decursin and its isomer decursinol angelate as potential inhibitors of human glutamate dehydrogenase activity through in silico and enzymatic assay screening

Glutaminolysis is a typical hallmark of malignant tumors across different cancers. Glutamate dehydrogenase (GDH, GLUD1) is one such enzyme involved in the conversion of glutamate to α-ketoglutarate. High levels of GDH are associated with numerous diseases and is also a prognostic marker for predicting metastasis in colorectal cancer. Therefore, inhibiting GDH can be a crucial therapeutic target. Here in this study, we performed molecular docking analysis of 8 different plants derived single compounds collected from pubChem database for screening and selected decursin (DN) and decursinol angelate (DA). We performed molecular dynamics simulation (MD), monitored the stability, interaction for protein and docked ligand at 50 ns, and evaluated the molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) free energy calculation on the twoselected compounds along with a standard inhibitor epigallocatechin gallate (EGCG) as reference. The final results showed the formation of stable hydrogen bond interactions by DN and DA in the residues of R400 and Y386 at the ADP activation site of GDH, which was important for the selective inhibition of GDH activity. Additionally, the total binding energy of DN and DA were −115.5 kJ/mol and −106.2 kJ/mol, which was higher than the standard reference GDH inhibitor EGCG (−92.8 kJ/mol). Furthermore, biochemical analysis for GDH inhibition substantiated our computational results and established DN and DA as novel GDH inhibitor. The percentage of IC50 inhibition for DN and DA were 1.035 μM and 1.432 μM. Conclusively, DN and DA can be a novel therapeutic drug for inhibition of glutamate dehydrogenase.

Arabidopsis thaliana sirtuins control proliferation and glutamate dehydrogenase activity

Sirtuins are part of a gene family of NAD-dependent deacylases that act on histone and non-histone proteins and control a variety of activities in all living organisms. Their roles are mainly related to energy metabolism and include lifetime regulation, DNA repair, stress resistance, and proliferation. A large amount of knowledge concerning animal sirtuins is available, but data about their plant counterparts are scarce. Plants possess few sirtuins that have, like in animals, a recognized role in stress defense and metabolism regulation. However, engagement in proliferation control, which has been demonstrated for mammalian sirtuins, has not been reported for plant sirtuins so far. In this work, srt1 and srt2 Arabidopsis mutant seedlings have been used to evaluate in vivo the role of sirtuins in cell proliferation and regulation of glutamate dehydrogenase, an enzyme demonstrated to be involved in the control of cell cycle in SIRT4-defective human cells. Moreover, bioinformatic analyses have been performed to elucidate sequence, structure, and function relationships between Arabidopsis sirtuins and between each of them and the closest mammalian homolog. We found that cell proliferation and GDH activity are higher in mutant seedlings, suggesting that both sirtuins exert a physiological inhibitory role in these processes. In addition, mutant seedlings show plant growth and root system improvement, in line with metabolic data. Our data also indicate that utilization of an easy to manipulate organism, such as Arabidopsis plant, can help to shed light on the molecular mechanisms underlying the function of genes present in interkingdom species.

Effect of glycine combined with urea on the absorption, transportation and transformation of nitrogen in wild peach seedlings (Amygdalu spersica (L.) Batsch)

The mechanism of how glycine combined with urea affects the development of peach trees through nitrogen absorption and metabolism is still unclear. In this study, we investigated the transformation of nitrogen in soil, the nitrogen absorption in roots, and the transformation and transportation of nitrogen in the peach tree at different treatment (no nitrogen fertilizer (CK), urea (T1), glycine (T2), glycine:urea (1:1) (T3), glycine:urea (3:7) (T4), and glycine:urea (7:3) (T5)). Compared with CK, the total dry weight of T5 and T2 was significantly increased by 22.95% and 19.09%, while it was lower at T3. The root activity of T5 was significantly higher than CK by 169.27%. The residual NH4+ in the soil was consistent with the application ratio of urea, while the conversion of NO3− was promoted at T5. Under T5, soil amoA and nifH abundance was higher, while the nirK abundance was higher under T1 and T3. In addition, urease activity was higher at T1, while phosphatase, catalase, and dehydrogenase activities were higher at T5. The NO3− content in the root was higher at T5. In roots, the activities of GDH, GOGAT and GS were significantly increased at T5 and T2, while the NR activities of T5 and T1 were significantly increased by 34.60% and 33.64% compared with CK. The aspartic acid and glutamic acid were the main forms of nitrogen transport in wild peaches. PLS-PM analysis showed that nitrogen use efficiency had a very significant direct positive effect on the biomass of peach trees.

Analysis of Relative Expression of Key Enzyme Genes and Enzyme Activity in Nitrogen Metabolic Pathway of Two Genotypes of Potato (Solanum tuberosum L.) under Different Nitrogen Supply Levels

Nitrogen (N) plays an important role in the growth cycle of the potato, and is an important guarantee of yield and quality. Rational N application is one of the key ways to improve a crop’s high and stable yield and economic and environmental benefits. The N nutrition level of potato regulates the gene expression of enzymes related to the N metabolism pathway, which shows the change of the activity of key enzymes in N metabolism, and finally realizes the regulation of N absorption and utilization. In this study, the key enzyme genes and enzyme activity of different genotypes of potato under different N supply treatments were identified, which laid a foundation for further exploration of the functions of each gene in the potato N metabolism pathway and provided theoretical basis for rational N application. The results showed that the relative expression levels of StNRT1.5, StNR, StNiR in leaves, StNRT2.5, StNRT2.4, StGS1-2 in roots and StNRT2.7, StGS1-3, StGS2, StGS1-4, StFd-GOGAT in leaves and roots showed that the levels of N-inefficient potato Atlantic (A) were higher than the N-efficient potato Yanshu 4 (Y), while the relative expression levels of StNRT2.5, StGS1-2 in leaves, StNRT1.5StNR, StNiR in roots and StGDH, StNADH-GOGAT in leaves and roots showed that levels in Yanshu 4 (Y) were higher than in Atlantic (A). At the same time, we especially found that the GDH activity in the leaves of the two genotypes of potato were higher at low N levels. Additionally, the activity of NR and NiR, and the activity of GS and GOGAT were correlated. In addition, the changes of key enzymes in different N metabolism showed a certain continuity with the advancement of growth and development, and some gene expression rules and enzyme activity changes also showed a certain consistency.