Activities Of Cycle Enzymes Research Articles

The role of endogenous nitric oxide in salicylic acid-induced up-regulation of ascorbate-glutathione cycle involved in salinity tolerance of pepper (Capsicum annuum L.) plants

An experimentation was carried out to appraise whether or not nitric oxide (NO) contributes to salicylic acid (SA)-induced salinity tolerance particularly by regulating ascorbate-glutathione (AsA-GSH) cycle. Before starting salinity stress (SS), SA (0.5 mM) was sprayed to the foliage of plants once every other day for a week and then seedlings were grown under control or SS (100 mM NaCl), for five weeks. Salinity stress enhanced the AsA-GSH cycle-related enzymes, glutathione reductase (GR), ascorbate peroxidase (APX), and dehydroascorbate reductase (DHAR), and monodehydroascorbate reductase (MDHAR). Furthermore, SS caused substantial decreases in plant physiological-related traits such as leaf potassium (K) contents, K+/Na+ ratio, the ratios of reduced ascorbate/dehydroascorbic acid (AsA/DHA) and reduced glutathione/oxidized glutathione (GSH/GSSG), but in contrast, significant increases occurred in leaf hydrogen peroxide, malondialdehyde, electron leakage, proline, the premier antioxidant enzymes’ activities, Na+ and NO. SA reduced leaf Na+ content and oxidative stress-related traits, but improved all earlier-mentioned traits compared with those in plants treated with SS alone. All positive effects of SA were eliminated by NO scavenger, 0.1 mM 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1- oxyl-3-oxide (c-PTIO) by reducing NO, suggesting that NO produced by SA up-regulated the activities of AsA-GSH cycle and antioxidant enzymes, so it could play a central function as a signal molecule in salt tolerance of pepper plants.

3-Hydroxy-3-Methylglutaric Acid Impairs Redox and Energy Homeostasis, Mitochondrial Dynamics, and Endoplasmic Reticulum-Mitochondria Crosstalk in Rat Brain.

3-Hydroxy-3-methylglutaryl-CoA lyase (HL) deficiency is a neurometabolic disorder characterized by predominant accumulation of 3-hydroxy-3-methylglutaric acid (HMG) in tissues and biological fluids. Patients often present in the first year of life with metabolic acidosis, non-ketotic hypoglycemia, hypotonia, lethargy, and coma. Since neurological symptoms may be triggered or worsened during episodes of metabolic decompensation, which are characterized by high urinary excretion of organic acids, this study investigated the effects of HMG intracerebroventricular administration on redox homeostasis, citric acid cycle enzyme activities, dynamics (mitochondrial fusion and fission), and endoplasmic reticulum (ER)-mitochondria crosstalk in the brain of neonatal rats euthanized 1 (short term) or 20 days (long term) after injection. HMG induced lipid peroxidation and decreased the activities of glutathione peroxidase (GPx) and citric acid cycle enzymes, suggesting bioenergetic and redox disruption, 1 day after administration. Levels of VDAC1, Grp75, and mitofusin-1, proteins involved in ER-mitochondria crosstalk and mitochondrial fusion, were increased by HMG. Furthermore, HMG diminished synaptophysin levels and tau phosphorylation, and increased active caspase-3 content, indicative of cell damage. Finally, HMG decreased GPx activity and synaptophysin levels, and changed MAPK phosphorylation 20 days after injection, suggesting that long-term toxicity is further induced by this organic acid. Taken together, these data show that HMG induces oxidative stress and disrupts bioenergetics, dynamics, ER-mitochondria communication, and signaling pathways in the brain of rats soon after birth. It may be presumed that these mechanisms underlie the onset and progression of symptoms during decompensation occurring in HL-deficient patients during the neonatal period.

Effects of 1-methylcyclopropene on ascorbate-glutathione cycle enzyme activities of postharvest Dendrobium ‘Khao Sanan’

Effects of 1-methylcyclopropene on ascorbate-glutathione cycle enzyme activities of postharvest <i>Dendrobium</i> ‘Khao Sanan’

Tricarboxylic acid cycle enzyme activities in a mouse model of methylmalonic aciduria

Methylmalonic acidemia (MMA) is a propionate pathway disorder caused by dysfunction of the mitochondrial enzyme methylmalonyl-CoA mutase (MMUT). MMUT catalyzes the conversion of methylmalonyl-CoA to succinyl-CoA, an anaplerotic reaction which feeds into the tricarboxylic acid (TCA) cycle. As part of the pathological mechanisms of MMA, previous studies have suggested there is decreased TCA activity due to a “toxic inhibition” of TCA cycle enzymes by MMA related metabolites, in addition to reduced anaplerosis. Here, we have utilized mitochondria isolated from livers of a mouse model of MMA (Mut-ko/ki) and their littermate controls (Ki/wt) to examine the amounts and enzyme functions of most of the TCA cycle enzymes. We have performed mRNA quantification, protein semi-quantitation, and enzyme activity quantification for TCA cycle enzymes in these samples. Expression profiling showed increased mRNA levels of fumarate hydratase in the Mut-ko/ki samples, which by contrast had reduced protein levels as detected by immunoblot, while all other mRNA levels were unaltered. Immunoblotting also revealed decreased protein levels of 2-oxoglutarate dehydrogenase and malate dehydrogenase 2. Interesting, the decreased protein amount of 2-oxoglutarate dehydrogenase was reflected in decreased activity for this enzyme while there is a trend towards decreased activity of fumarate hydratase and malate dehydrogenase 2. Citrate synthase, isocitrate dehydrogenase 2/3, succinyl-CoA synthase, and succinate dehydrogenase are not statistically different in terms of quantity of enzyme or activity. Finally, we found decreased activity when examining the function of methylmalonyl-CoA mutase in series with succinate synthase and succinate dehydrogenase in the Mut-ko/ki mice compared to their littermate controls, as expected. This study demonstrates decreased activity of certain TCA cycle enzymes and by corollary decreased TCA cycle function, but it supports decreased protein quantity rather than “toxic inhibition” as the underlying mechanism of action. SummaryMethylmalonic acidemia (MMA) is an inborn metabolic disorder of propionate catabolism. In this disorder, toxic metabolites are considered to be the major pathogenic mechanism for acute and long-term complications. However, despite optimized therapies aimed at reducing metabolite levels, patients continue to suffer from late complications, including metabolic stroke and renal insufficiency. Since the propionate pathway feeds into the tricarboxylic acid (TCA) cycle, we investigated TCA cycle function in a constitutive MMA mouse model. We demonstrated decreased amounts of the TCA enzymes, Mdh2 and Ogdh as semi-quantified by immunoblot. Enzymatic activity of Ogdh is also decreased in the MMA mouse model compared to controls. Thus, when the enzyme amounts are decreased, we see the enzymatic activity also decreased to a similar extent for Ogdh. Further studies to elucidate the structural and/or functional links between the TCA cycle and propionate pathways might lead to new treatment approaches for MMA patients.

Ammonia Scavenging Prevents Progression of Fibrosis in Experimental Nonalcoholic Fatty Liver Disease.

In nonalcoholic fatty liver disease (NAFLD), fibrosis is the most important factor contributing to NAFLD-associated morbidity and mortality. Prevention of progression and reduction in fibrosis are the main aims of treatment. Even in early stages of NAFLD, hepatic and systemic hyperammonemia is evident. This is due to reduced urea synthesis; and as ammonia is known to activate hepatic stellate cells, we hypothesized that ammonia may be involved in the progression of fibrosis in NAFLD. In a high-fat, high-cholesterol diet-induced rodent model of NAFLD, we observed a progressive stepwise reduction in the expression and activity of urea cycle enzymes resulting in hyperammonemia, evidence of hepatic stellate cell activation, and progressive fibrosis. In primary, cultured hepatocytes and precision-cut liver slices we demonstrated increased gene expression of profibrogenic markers after lipid and/or ammonia exposure. Lowering of ammonia with the ammonia scavenger ornithine phenylacetate prevented hepatocyte cell death and significantly reduced the development of fibrosis both in vitro in the liver slices and in vivo in a rodent model. The prevention of fibrosis in the rodent model was associated with restoration of urea cycle enzyme activity and function, reduced hepatic ammonia, and markers of inflammation. The results of this study suggest that hepatic steatosis results in hyperammonemia, which is associated with progression of hepatic fibrosis. Reduction of ammonia levels prevented progression of fibrosis, providing a potential treatment for NAFLD.

Increased CO2 and light intensity regulate growth and leaf gas exchange in tomato.

Carbon dioxide concentration (CO2 ) and light intensity are known to play important roles in plant growth and carbon assimilation. Nevertheless, the underlying physiological mechanisms have not yet been fully explored. Tomato seedlings (Solanum lycopersicum Mill. cv. Jingpeng No. 1) were exposed to two levels of CO2 and three levels of light intensity and the effects on growth, leaf gas exchange and water use efficiency were investigated. Elevated CO2 and increased light intensity promoted growth, dry matter accumulation and pigment concentration and together the seedling health index. Elevated CO2 had no significant effect on leaf nitrogen content but did significantly upregulate Calvin cycle enzyme activity. Increased CO2 and light intensity promoted photosynthesis, both on a leaf-area basis and on a chlorophyll basis. Increased CO2 also increased light-saturated maximum photosynthetic rate, apparent quantum efficiency and carboxylation efficiency and, together with increased light intensity, it raised photosynthetic capacity. However, increased CO2 reduced transpiration and water consumption across different levels of light intensity, thus significantly increasing both leaf-level and plant-level water use efficiency. Among the range of treatments imposed, the combination of increased CO2 (800 µmol CO2 mol-1 ) and high light intensity (400 µmol m-2 s-1 ) resulted in optimal growth and carbon assimilation. We conclude that the combination of increased CO2 and increased light intensity worked synergistically to promote growth, photosynthetic capacity and water use efficiency by upregulation of pigment concentration, Calvin cycle enzyme activity, light energy use and CO2 fixation. Increased CO2 also lowered transpiration and hence water usage.

Intense Light-Mediated Circadian Cardioprotection via Transcriptional Reprogramming of the Endothelium

SUMMARYConsistent daylight oscillations and abundant oxygen availability are fundamental to human health. Here, we investigate the intersection between light-sensing (Period 2 [PER2]) and oxygen-sensing (hypoxia-inducible factor [HIF1A]) pathways in cellular adaptation to myocardial ischemia. We demonstrate that intense light is cardioprotective via circadian PER2 amplitude enhancement, mimicking hypoxia-elicited adenosine- and HIF1A-metabolic adaptation to myocardial ischemia under normoxic conditions. Whole-genome array from intense light-exposed wild-type or Per2−/− mice and myocardial ischemia in endothelial-specific PER2-deficient mice uncover a critical role for intense light in maintaining endothelial barrier function via light-enhanced HIF1A transcription. A proteomics screen in human endothelia reveals a dominant role for PER2 in metabolic reprogramming to hypoxia via mitochondrial translocation, tricarboxylic acid (TCA) cycle enzyme activity regulation, and HIF1A transcriptional adaption to hypoxia. Translational investigation of intense light in human subjects identifies similar PER2 mechanisms, implicating the use of intense light for the treatment of cardiovascular disease.

Exogenous spermidine enhances expression of Calvin cycle genes andphotosynthetic efficiency in sweet sorghum seedlings under salt stress

Salinity adversely affects plants resulting in disruption to plant growth and physiology. Previously, it has been shown that these negative effects can be alleviated by various exogenous polyamines. However, the role of spermidine (Spd) in conferring salinity tolerance in sorghum is not well documented. The effect of exogenous Spd on the responses of sweet sorghum (Sorghum bicolor L.) seedlings to salt stress (150 mM NaCl) was investigated by measuring photosynthetic carbon assimilation, Calvin cycle enzyme activities, and the the expression of respective genes. Application of 0.25 mM Spd alleviated the negative effects of salt stress on efficiency of photosystem II and CO2 assimilation and increased the activities of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and aldolase. Salt stress significantly lowered the transcriptions of genes encoding Rubisco large subunit, Rubisco small subunit, 3-phosphoglyceric acid kinase, glyceraldehyde-3-phosphate dehydrogenase, triose-3-phosphate isomerase, fructose-1,6-bisphosphate aldolase, fructose-1,6-bisphosphate phosphatase, and sedoheptulose-1,7-bisphosphatase. However, transcriptions of genes encoding phosphoribokinase and Rubisco were up-regulated. The Spd application enhanced expressions of most of these genes. It appears Spd conferred salinity tolerance to sweet sorghum seedlings by enhancing photosynthetic efficiency through regulation of gene expressions and activities of key CO2 assimilation enzymes.

Oxidative attack during temperature fluctuation challenge compromises liver protein homeostasis of a temperate fish model.

Seasonal variations in water temperature are a natural stressor of temperate fish that affect growth performance and metabolism globally. Gilthead sea bream is one of the most economically interesting species in the Mediterranean; but its liver metabolism is affected by the cold season. However, the effects of cold on protein turnover mechanisms have hardly been studied. Here, we study the relationship between liver oxidative status and protein homeostasis pathways during a 50-day low temperature period at 14 °C, and subsequent recovery at two times: 7 days (early recovery) and 30 days (late recovery). Liver redox status was determined by measuring oxidised lipids and proteins, the glutathione redox cycle and major antioxidant enzymes activities. Protein turnover was analysed via liver protein expression of HSP70 and HSP90; proteasome 26S subunits and polyubiquitination, as markers of the ubiquitin-proteasome system (UPS); and cathepsin D, as a lysosomal protease. Low temperature exposure depressed antioxidant enzyme activities, affecting the glutathione redox cycle and reducing total glutathione levels. Both the UPS and lysosomal pathways were also depressed and consequently, oxidised protein accumulated in liver. Interestingly, both protein oxidation and polyubiquitination tagging depended on protein molecular weight. Despite all these alterations, temperature recovery reverted most consequences of the cold at different rates: with delayed recovery of total glutathione levels and oxidised protein degradation with respect to enzyme activities recovery. All these findings demonstrate that protein liver homeostasis is compromised after chronic cold exposure and could be the cause of liver affectations reported in aquaculture of temperate fish.

Synthesis and Biological Assessment of Pyrrolobenzoxazine Scaffold as a Potent Antioxidant.

Reduction of mitochondrial oxidative stress-mediated diseases is an important pharmaceutical objective in recent biomedical research. In this context, a series of novel pyrrolobenzoxazines (PyBs) framework with enormous diversity (compounds 5a-w) was synthesized by employing a low-temperature greener pathway, and antioxidant property of the synthesized compounds was successfully demonstrated on preclinical model goat heart mitochondria, in vitro. Copper-ascorbate (Cu-As) was utilized as an oxidative stress generator. Out of screened PyBs, the compound possessing -OH and -OMe groups on benzene nucleus along with pyrrolobenzoxazine core moiety (compound 5w) displayed magnificent antioxidant property with a minimum effective dose of 66 μM during the biochemical assessment. The ameliorative effect of synthesized pyrrolobenzoxazine moiety on levels of biomarkers of oxidative stress, antioxidant enzyme, activities of Krebs cycle and respiratory chain enzymes, mitochondrial morphology, and Ca2+ permeability of mitochondrial membrane was investigated in the presence of Cu-As. Furthermore, the binding mode of Cu-As by compound 5w was explored successfully using isothermal titration calorimetry (ITC) analysis.

Regulation of chromium toxicity tolerance in tomato and brinjal by calcium and sulfur through nitric oxide: Involvement of enzymes of sulfur assimilation and the ascorbate-glutathione cycle

In soils, toxic metals pose negative impact on crop productivity worldwide. This warrants need of some cost effective and eco-friendly strategies which could mitigate/manage metal toxicity in crop plants for sustained and increased crop productivity. In the past years, exogenous application of nutrient elements has attracted wider attention in mitigating negative consequences of varied abiotic stresses with a much focus on Cd toxicity. However, potential of nutrients like calcium (Ca) and sulfur (S) in mitigating hexavalent chromium [Cr(VI)] toxicity in vegetable crops together with their mechanisms of action are still not known. Therefore, in this study potential of Ca and S in mitigating Cr(VI) toxicity in tomato and brinjal along with their mode of action have been explored. Cr(VI) significantly decreased leaf area, photosynthetic rate, sub-stomatal CO2, stomatal conductance and transpiration rate and cell viability due to enhanced intracellular accumulation of Cr(VI). Cr(VI) enhanced generation of reactive oxygen species (ROS) i.e. superoxide radical (O2•̄) and hydrogen peroxide (H2O2) that leads to the enhanced protein oxidation and decreased membrane stability index (MSI) as a result of down-regulation in S assimilation and ascorbate-glutathione cycle (AsA-GSH cycle; enzymes activity as well as relative gene expression). The results also show that Cr(VI)-mediated negative impacts were accompanied by declined endogenous nitric oxide (NO). However, additional supply of either Ca or S with Cr(VI) significantly ameliorated Cr(VI) toxicity by down-regulating accumulation of Cr(VI) and ROS as a result of up-regulation in S assimilation and AsA-GSH cycle (enzymes activity as well as relative gene expression) which collectively protect photosynthetic characteristics and cell viability. Interestingly addition of Nω-nitro-L-arginine methylester (L-NAME, an inhibitor of nitric oxide synthase) with Cr(VI) further increased Cr(VI) toxicity in both vegetables while addition of sodium nitroprusside (a NO donor) reversed this response. Thus, overall results showed that NO is an essential component in Ca and S-mediated mitigation of Cr(VI) toxicity in tomato and brinjal seedlings.

Ursolic acid modulates MMPs, collagen-I, α-SMA, and TGF-β expression in isoproterenol-induced myocardial infarction in rats.

In the present study, the modulatory effect of ursolic acid (UA) on cardiac fibrosis and mitochondrial and lysosomal enzymes activity in isoproterenol-induced myocardial infarction (MI) in rats were examined. Isoproterenol hydrochloride (ISO; 85 mg/kg body weight) was administered subcutaneously for first two consecutive days. ISO-induced MI in rats significantly decreased the activities of mitochondrial tricarboxylic acid cycle enzymes and respiratory chain enzymes while increased the activities of lysosomal glycohydrolases and cathepsins. The expression of matrix metalloproteinase 2 (MMP-2), MMP-9, collagen type I, α-smooth muscle actin (α-SMA), and transforming growth factor-β (TGF-β) were upregulated in ISO-induced MI in rats. UA administration to rats showed increased activities of mitochondrial tricarboxylic acid cycle enzymes and respiratory chain enzymes and decreased activities of lysosomal glycohydrolases and cathepsins in ISO-induced rats. Furthermore, expression of MMP-2, MMP-9, collagen type I, α-SMA, and TGF-β downregulated in UA-administered rats. Thus, our results demonstrate that UA has an anti-fibrotic effect and attenuates the mitochondrial and lysosomal dysfunction in ISO-induced MI in rats.

The Food Energy/Protein Ratio Regulates the Rat Urea Cycle but Not Total Nitrogen Losses.

Nitrogen balance studies have shown that a portion of the N ingested but not excreted is not accounted for. We compared several diets (standard, high-fat, high-protein, and self-selected cafeteria) to determine how diet-dependent energy sources affect nitrogen handling, i.e., the liver urea cycle. Diet components and rat homogenates were used for nitrogen, lipid, and energy analyses. Plasma urea and individual amino acids, as well as liver urea cycle enzyme activities, were determined. Despite ample differences in N intake, circulating amino acids remained practically unchanged in contrast to marked changes in plasma urea. The finding of significant correlations between circulating urea and arginine-succinate synthase and lyase activities supported their regulatory role of urea synthesis, the main N excretion pathway. The cycle operation also correlated with the food protein/energy ratio, in contraposition to total nitrogen losses and estimated balance essentially independent of dietary energy load. The different regulation mechanisms observed have potentially important nutritional consequences, hinting at nitrogen disposal mechanisms able to eliminate excess nitrogen under conditions of high availability of both energy and proteins. Their operation reduces urea synthesis to allow for a safe (albeit unknown) mechanism of N/energy excess accommodation.

Chromomycin, an antibiotic produced by Streptomyces flaviscleroticus might play a role in the resistance to oxidative stress and is essential for viability in stationary phase.

The well-known role of antibiotics in killing sensitive organisms has been challenged by the effects they exert at subinhibitory concentrations. Unfortunately, there are very few published reports on the advantages these molecules may confer to their producers. This study describes the construction of a genetically verified deletion mutant of Streptomyces flaviscleroticus unable to synthesize chromomycin. This mutant was characterized by a rapid loss of viability in stationary phase that was correlated with high oxidative stress and altered antioxidant defences. Altered levels of key metabolites in the mutant signalled a redistribution of the glycolytic flux toward the PPP to generate NADPH to fight oxidative stress as well as reduction of ATP-phosphofructokinase and Krebs cycle enzymes activities. These changes were correlated with a shift in the preference for carbon utilization from glucose to amino acids. Remarkably, chromomycin at subinhibitory concentration increased longevity of the non-producer and restored most of the phenotypic features' characteristic of the wild type strain. Altogether these observations suggest that chromomycin may have antioxidant properties that would explain, at least in part, some of the phenotypes of the mutant. Our observations warrant reconsideration of the secondary metabolite definition and raise the possibility of crucial roles for their producers.

Exogenous silicon alters organic acid production and enzymatic activity of TCA cycle in two NaCl stressed indica rice cultivars

The activities of TCA cycle enzymes viz., pyruvate dehydrogenase, citrate synthase, isocitrate dehydrogenase, succinate dehydrogenase and malate dehydrogenase as well as levels of different organic acids viz., pyruvic acid, citric acid, succinic acid and malic acid were studied in two rice cultivars viz. cv. Nonabokra and cv. MTU 1010 differing in salt tolerance grown under 25, 50 and 100 mM NaCl salinity levels. A contrasting response to salt stress on enzyme activities of TCA cycle and accumulation of organic acid was observed between two cultivars during twenty-one days period of study. Salinity caused enhanced organic acid production and increase in all five enzyme activities in cv. Nonabokra whereas in cv. MTU 1010 decrease in both organic acid production and enzymes activities were noted. Joint application of exogenous silicon along with NaCl, altered the organic acids levels and activities of enzymes in both cultivars of rice seedlings conferring tolerance against salt induced stress. Rice cv. MTU 1010 showed better response to exogenous silicon on parameters tested compared to cv. Nonabokra.

Biochemical and Physiological Effects of Some Organic and Inorganic Chemical Agents in Capsicum spp.

Cultivation of vegetables is expected to increase in order to meet the demands of the expanding populations of the globe. Meanwhile, anthropic activities increase the concentrations of various chemical compounds in aquatic and terrestrial habitats. For productivity and food safety reasons, assessments of effects of pesticides and metals on crops should be performed. In the current paper, the presence of some heavy metals and a pesticide compound in the substrate altered the levels of some Krebs cycle enzymes activities in pepper plants cultivated in controlled conditions. The photosynthetic apparatus of the same plants appeared relatively unaffected, while the potential/actual soil dehydrogenases ratios were increased in all treatments.

Catalase, superoxide dismutase and ascorbate-glutathione cycle enzymes confer drought tolerance of Amaranthus tricolor

The study was performed to explore physiological, non-enzymatic and enzymatic detoxification pathways of reactive oxygen species (ROS) in tolerance of Amaranthus tricolor under drought stress. The tolerant genotype VA13 exhibited lower reduction in growth, photosynthetic pigments, relative water content (RWC) and negligible increment in electrolyte leakage (EL), lower increment in proline, guaiacol peroxidase (GPOX) activity compared to sensitive genotype VA15. This genotype also had higher catalase (CAT), superoxide dismutase (SOD), remarkable and dramatic increment in ascorbate-glutathione content, ascorbate-glutathione redox and ascorbate-glutathione cycle enzymes activity compared to sensitive genotype VA15. The negligible increment of ascorbate-glutathione content, ascorbate-glutathione redox and ascorbate-glutathione cycle enzymes activities and dramatic increment in malondialdehyde (MDA), hydrogen peroxide (H2O2) and EL were observed in the sensitive genotype VA15. SOD contributed superoxide radical dismutation and CAT contributed H2O2 detoxification in both sensitive and tolerant varieties, however, these had a great contribution in the tolerant variety. Conversely, proline and GPOX accumulation were higher in the sensitive variety compared to the tolerant variety. Increase in ascorbate-glutathione cycle enzymes activities, CAT, ascorbate-glutathione content, SOD, and ascorbate-glutathione redox clearly evident that CAT, ascorbate-glutathione cycle and SOD played a significant activity in ROS detoxification of tolerant A. tricolor variety.

Silicon induced mitigation of TCA cycle and GABA synthesis in arsenic stressed wheat (Triticum aestivum L.) seedlings

The metalloid arsenic (As) is considered to be biologically non-essential and major environmental concern to all organisms. The aim of our study was to evaluate whether silicon (Si) has ameliorative effects on growth, respiratory cycle and Gamma-Aminobutyric acid (GABA) synthesis in wheat (Triticum aestivum L. cv PBW 343) seedlings under arsenic stress. The experiments were performed in a completely randomized design with three repeats and two replicates for each treatment. The seedlings were subjected to different doses of arsenate As (V) (25, 50 and 100 μM) with or without silicate (5 mM) in modified Hoagland's nutrient media for 21 days. Arsenate stress decreased seedling growth, activities of respiratory cycle enzymes particularly dehydrogenases while enhanced the levels of oxidative stress markers, Krebs cycle intermediates and GABA including activities of enzymes associated with GABA synthesis. Arsenate treatment disturbed the phosphate contents and impaired the respiratory process. Irrespective of arsenate concentrations, silicate administration substantially modulated the toxic effects of the metalloid in the test cultivar. Thus, silicon supplementation may emerge as a beneficial strategy to reduce potential health risks and might help to mitigate arsenic- induced stress in wheat seedlings.

Exogenous silicon alters ascorbate-glutathione cycle in two salt-stressed indica rice cultivars (MTU 1010 and Nonabokra).

Silicon is widely available in soil and is known to mitigate both biotic and abiotic stress in plants. Very low doses of silicon are becoming increasingly essential in rice for biofortification and preventing water loss. Soil salinity is a matter of grave concern in various parts of the world, and silicon is a suitable candidate to mitigate salinity-induced stress of important plants in affected areas. The present study investigates the protective capability of exogenously applied silicon in ameliorating NaCl-induced toxicity in two rice (Oryza sativa L.) cultivars, the salt-sensitive MTU 1010, and salt-tolerant Nonabokra. Rice seedlings were treated with three doses of NaCl (25, 50, and 100mM), initially alone and subsequently in combination with 2mM sodium silicate (Na2SiO3, 9H2O). After 21days, these plants were examined to determine levels of reduced glutathione, ascorbic acid, cysteine, and activities of different enzymes involved in the ascorbate-glutathione cycle, viz., glutathione reductase (GR), ascorbate peroxidase (APX), glutathione peroxidase (GPx), and glutathione S-transferase (GST). Though ROS levels increased in both the cultivars with increasing NaCl concentrations, cv. MTU 1010 accumulated comparatively higher amounts. A differential response of NaCl-induced toxicity on the two cultivars was observed with respect to the various enzymatic and non-enzymatic antioxidants. APX and GST activities, as well as, cysteine contents, increased concomitantly with salt concentrations, whereas GR activity declined at increasing salt concentrations, in both cultivars. Activity of GPx increased in cv. Nonabokra but declined in cv. MTU 1010, under similar NaCl concentrations. Reduced glutathione (GSH) contents decreased in both cultivars, whereas ascorbate contents declined in only the sensitive cultivar. Application of silicon, along with NaCl, in the test seedlings of both the cultivars, reduced ROS accumulation and boosted antioxidant defense mechanism, through enhancing ascorbate and GSH levels, and activities of ascorbate-glutathione cycle enzymes as well. However, amelioration of salt-induced damages in the sensitive cv. MTU 1010 was more pronounced upon silicon administration, than the tolerant cv. Nonabokra. Thus, cv. MTU 1010 was found to be more responsive to applied silicon. Hence, this study was instrumental in realizing a successful strategy in silicon-mediated amelioration of salinity stress in plants.

The Aspergillus nidulans Pyruvate Dehydrogenase Kinases Are Essential To Integrate Carbon Source Metabolism.

The pyruvate dehydrogenase complex (PDH), that converts pyruvate to acetyl-coA, is regulated by pyruvate dehydrogenase kinases (PDHK) and phosphatases (PDHP) that have been shown to be important for morphology, pathogenicity and carbon source utilization in different fungal species. The aim of this study was to investigate the role played by the three PDHKs PkpA, PkpB and PkpC in carbon source utilization in the reference filamentous fungus Aspergillus nidulans, in order to unravel regulatory mechanisms which could prove useful for fungal biotechnological and biomedical applications. PkpA and PkpB were shown to be mitochondrial whereas PkpC localized to the mitochondria in a carbon source-dependent manner. Only PkpA was shown to regulate PDH activity. In the presence of glucose, deletion of pkpA and pkpC resulted in reduced glucose utilization, which affected carbon catabolite repression (CCR) and hydrolytic enzyme secretion, due to de-regulated glycolysis and TCA cycle enzyme activities. Furthermore, PkpC was shown to be required for the correct metabolic utilization of cellulose and acetate. PkpC negatively regulated the activity of the glyoxylate cycle enzyme isocitrate lyase (ICL), required for acetate metabolism. In summary, this study identified PDHKs important for the regulation of central carbon metabolism in the presence of different carbon sources, with effects on the secretion of biotechnologically important enzymes and carbon source-related growth. This work demonstrates how central carbon metabolism can affect a variety of fungal traits and lays a basis for further investigation into these characteristics with potential interest for different applications.