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A molecular switch based on the metastable radical anion derived from a substituted heteroaryl quinone is described. Pyrrolyl quinone thiocyanate (PQ 9) showed an interaction with the fluoride anion that was visible to the naked eye and quantified by UV/vis and 1H and 13â C NMR. The metastable quinoid species formed by the interaction with F- ("ON" state) showed a molecular switching effect autocontrolled by the presence of ascorbate ("OFF" state) and back to the "ON" state by an autooxidation process, measured by visible and UV/vis spectroscopy. Due to its out-of-equilibrium properties and the exchange of matter and energy, a dissipative structural behaviour is proposed. Considering its similarity to the mechanism of coenzyme Q in oxidative phosphophorylation, PQ 9 was evaluated on Saccharomyces cerevisiae mitochondrial function for inhibition of complexes II, III and IV, reactive oxygen species (ROS) production, catalase activity and lipid peroxidation. The results showed that PQ 9 inhibited complex III activity as well as the activity of all electron transport chain (ETC) complexes. In addition, PQ 9 reduced ROS production and catalase activity in yeast. The results suggest that PQ 9 may have potential applications as a new microbicidal compound by inducing ETC dysfunction.
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This study evaluated the effect of different concentrations of alpha-tocopherol in gel form on fracture strength, hybrid layer formation, and microtensile bond strength of endodontically treated teeth bleached with 40% hydrogen peroxide (H2 O2 ). Sixty bovine incisors were randomized into one of six groups (n = 10 incisors per group) defined by the interventions carried out after endodontic treatment. In the control group, no additional intervention was carried out, while all teeth in the five intervention groups were bleached with 40% H2 O2 and subsequently treated with alpha-tocopherol at concentrations of 15% (15AT), 20% (20AT), or 25% (25AT), with 10% sodium ascorbate (10SA), or with nothing (40HP). Fracture strength was evaluated in a mechanical testing machine, hybrid layer formation was assessed using scanning electron microscopy, and bond strength was determined using microtensile bond-strength testing. Data were analyzed using Kruskal-Wallis and Dunn's tests. No statistically significant difference regarding fracture strength was observed among groups. Hybrid layer formation was greater in the 15AT group than in groups 40HP and 10SA. Teeth in groups 15AT, 20AT, and 25AT demonstrated higher bond strength than teeth in groups 40HP and 10SA. Alpha-tocopherol, preferably at 15%, effectively reverses the deleterious effects, of bleaching, on hybrid layer formation and bond strength to dentin.
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Colagem Dentária , Clareamento Dental , Dente não Vital , Bovinos , Animais , Antioxidantes/química , Antioxidantes/farmacologia , alfa-Tocoferol/farmacologia , Resistência à Flexão , Resinas Compostas/química , Ácido HipoclorosoRESUMO
We have previously shown that rice plants silenced for peroxisomal ascorbate peroxidase (OsAPX4-RNAi) display higher resilience to photosynthesis under oxidative stress and photorespiratory conditions. However, the redox mechanisms underlying that intriguing response remain unknown. Here, we tested the hypothesis that favorable effects triggered by peroxisomal APX deficiency on photosynthesis resilience under CAT inhibition are dependent on the intensity of photorespiration associated with the abundance of photosynthetic and redox proteins. Non-transformed (NT) and OsAPX4-RNAi silenced rice plants were grown under ambient (AC) or high CO2 (HC) conditions and subjected to 3-amino-1,2,4-triazole (3-AT)-mediated CAT activity inhibition. Photosynthetic measurements evidenced that OsAPX4-RNAi plants simultaneously exposed to CAT inhibition and HC lost the previously acquired advantage in photosynthesis resilience displayed under AC. Silenced plants exposed to environment photorespiration and CAT inhibition presented lower photorespiration as indicated by smaller Gly/Ser and Jo/Jc ratios and glycolate oxidase activity. Interestingly, when these silenced plants were exposed to HC and CAT-inhibition, they exhibited an inverse response compared to AC in terms of photorespiration indicators, associated with higher accumulation of proteins. Multivariate and correlation network analyses suggest that the proteomics changes induced by HC combined with CAT inhibition are substantially different between NT and OsAPX4-RNAi plants. Our results suggest that the intensity of photorespiration and peroxisomal APX-mediated redox signaling are tightly regulated under CAT inhibition induced oxidative stress, which can modulate the photosynthetic efficiency, possibly via a coordinated regulation of protein abundance and rearrangement, ultimately triggered by crosstalk involving H2O2 levels related to CAT and APX activities in peroxisomes.
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Oryza , Oryza/metabolismo , Peróxido de Hidrogênio/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fotossíntese , Estresse Oxidativo , Plantas/metabolismo , Ascorbato Peroxidases/metabolismoRESUMO
This study focuses on developing and evaluating two novel enantioselective biomimetic models for the active centers of oxidases (ascorbate oxidase and catalase). These models aim to serve as alternatives to enzymes, which often have limited action and a delicate nature. For the ascorbate oxidase (AO) model (compound 1), two enantiomers, S,S(+)cpse and R,R(-)cpse, were combined in a crystalline structure, resulting in a racemic compound. The analysis of their magnetic properties and electrochemical behavior revealed electronic transfer between six metal centers. Compound 1 effectively catalyzed the oxidation of ascorbic to dehydroascorbic acid, showing a 45.5% yield for the racemic form. This was notably higher than the enantiopure compounds synthesized previously and tested in the current report, which exhibited yields of 32% and 28% for the S,S(+)cpse and R,R(-)cpse enantiomers, respectively. This outcome highlights the influence of electronic interactions between metal ions in the racemic compound compared to pure enantiomers. On the other hand, for the catalase model (compound 2), both the compound and its enantiomer displayed polymeric properties and dimeric behavior in the solid and solution states, respectively. Compound 2 proved to be effective in catalyzing the oxidation of hydrogen peroxide to oxygen with a yield of 64.7%. In contrast, its enantiomer (with R,R(-)cpse) achieved only a 27% yield. This further validates the functional nature of the prepared biomimetic models for oxidases. This research underscores the importance of understanding and designing biomimetic models of metalloenzyme active centers for both biological and industrial applications. These models show promising potential as viable alternatives to natural enzymes in various processes.
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OBJECTIVE: This study evaluated the effects of antioxidants, 10% sodium ascorbate (SA) or 20% alpha-tocopherol (AT), after post-space irrigation with 2.5% sodium hypochlorite +17% EDTA (SH) or 1% peracetic acid (PA) on the adhesive interface after glass fiber post cementation. MATERIALS AND METHODS: Sixty bovine roots were endodontically treated. After preparation, the post-space was irrigated with SH or PA followed or not by the use of antioxidants (SA or AT) (n = 10). Push-out bond strength test, failure mode, and dentin penetrability analysis using confocal laser microscope were performed in the cervical, middle, and apical thirds. Data from bond strength and dentinal penetrability were evaluated by one-way ANOVA and Tukey post hoc test (p < 0.05). RESULTS: SH showed the lowest bond strength regardless of the third (p < 0.05). In apical third, mixed failure was the most incident in all groups. Only in the cervical third of the post-space, SH-AT provided the greatest tag extension of the cementation system into dentin (p < 0.05). However, in the middle and apical thirds, SH-AT, SH-SA, and PA-SA provided the largest tag extensions (p < 0.05), but similar to each other (p > 0.05). CONCLUSIONS: The use of antioxidants only favored bond strength when SH was used and dentin penetrability of the adhesive and conventional resin cementation, regardless of the solution used to irrigate the post-space. CLINICAL SIGNIFICANCE: The use of antioxidants (10% sodium ascorbate and 20% alpha-tocopherol) after post-space irrigation with sodium hypochlorite appears to increase the bond strength favoring the glass fiber post-cementation.
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Colagem Dentária , Técnica para Retentor Intrarradicular , Bovinos , Animais , Cimentação , Antioxidantes/farmacologia , Cimentos Dentários/química , Cimentos de Resina/química , alfa-Tocoferol , Hipoclorito de Sódio/química , Dentina , Ácido Ascórbico/farmacologia , Teste de MateriaisRESUMO
Studies on the role of nickel (Ni) in photosynthetic and antioxidant metabolism, as well as in flavonoid synthesis and biological fixation nitrogen in cowpea crop are scarce. The aim of this study was to elucidate the role of Ni in metabolism, photosynthesis and nodulation of cowpea plants. A completely randomized experiment was performed in greenhouse, with cowpea plants cultivated under 0, 0.5, 1, 2, or 3 mg kg-1 Ni, as Ni sulfate. In the study the following parameters were evaluated: activity of urease, nitrate reductase, superoxide dismutase, catalase and ascorbate peroxidase; concentration of urea, n-compounds, photosynthetic pigments, flavonoids, H2O2 and MDA; estimative of gas exchange, and biomass as plants, yield and weight of 100 seeds. At whole-plant level, Ni affected root biomass, number of seeds per pot, and yield, increasing it at 0.5 mg kg-1 and leading to inhibition at 2-3 mg kg-1 (e.g. number of seeds per pot and nodulation). The whole-plant level enhancement by 0.5 mg Ni kg-1 occurred along with increased photosynthetic pigments, photosynthesis, ureides, and catalase, and decreased hydrogen peroxide concentration. This study presents fundamental new insights regarding Ni effect on N metabolism, and nodulation that can be helpful to increase cowpea yield. Considering the increasing population and its demand for staple food, these results contribute to the enhancement of agricultural techniques that increase crop productivity and help to maintain human food security.
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Vigna , Humanos , Catalase/metabolismo , Vigna/metabolismo , Fixação de Nitrogênio , Níquel/farmacologia , Níquel/metabolismo , Peróxido de Hidrogênio/metabolismoRESUMO
A healing material must have desirable characteristics such as maintaining a physiological environment, protective barrier-forming abilities, exudate absorption, easy handling, and non-toxicity. Laponite is a synthetic clay with properties such as swelling, physical crosslinking, rheological stability, and drug entrapment, making it an interesting alternative for developing new dressings. This study evaluated its performance in lecithin/gelatin composites (LGL) as well as with the addition of maltodextrin/sodium ascorbate mixture (LGL MAS). These materials were applied as nanoparticles, dispersed, and prepared by using the gelatin desolvation method-eventually being turned into films via the solvent-casting method. Both types of composites were also studied as dispersions and films. Dynamic Light Scattering (DLS) and rheological techniques were used to characterize the dispersions, while the films' mechanical properties and drug release were determined. Laponite in an amount of 8.8 mg developed the optimal composites, reducing the particulate size and avoiding the agglomeration by its physical crosslinker and amphoteric properties. On the films, it enhanced the swelling and provided stability below 50 °C. Moreover, the study of drug release in maltodextrin and sodium ascorbate from LGL MAS was fitted to first-order and Korsmeyer-Peppas models, respectively. The aforementioned systems represent an interesting, innovative, and promising alternative in the field of healing materials.
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Vitamin C (L-ascorbic acid, AsA) is an essential compound with pleiotropic functions in many organisms. Since its isolation in the last century, AsA has attracted the attention of the scientific community, allowing the discovery of the L-galactose pathway, which is the main pathway for AsA biosynthesis in plants. Thus, the aim of this review is to analyze the genetic and biochemical strategies employed by plant cells for regulating AsA biosynthesis through the L-galactose pathway. In this pathway, participates eight enzymes encoded by the genes PMI, PMM, GMP, GME, GGP, GPP, GDH, and GLDH. All these genes and their encoded enzymes have been well characterized, demonstrating their participation in AsA biosynthesis. Also, have described some genetic and biochemical strategies that allow its regulation. The genetic strategy includes regulation at transcriptional and post-transcriptional levels. In the first one, it was demonstrated that the expression levels of the genes correlate directly with AsA content in the tissues/organs of the plants. Also, it was proved that these genes are light-induced because they have light-responsive promoter motifs (e.g., ATC, I-box, GT1 motif, etc.). In addition, were identified some transcription factors that function as activators (e.g., SlICE1, AtERF98, SlHZ24, etc.) or inactivators (e.g., SlL1L4, ABI4, SlNYYA10) regulate the transcription of these genes. In the second one, it was proved that some genes have alternative splicing events and could be a mechanism to control AsA biosynthesis. Also, it was demonstrated that a conserved cis-acting upstream open reading frame (5'-uORF) located in the 5'-untranslated region of the GGP gene induces its post-transcriptional repression. Among the biochemical strategies discovered is the control of the enzyme levels (usually by decreasing their quantities), control of the enzyme catalytic activity (by increasing or decreasing its activity), feedback inhibition of some enzymes (GME and GGP), subcellular compartmentation of AsA, the metabolon assembly of the enzymes, and control of AsA biosynthesis by electron flow. Together, the construction of this basic knowledge has been establishing the foundations for generating genetically improved varieties of fruits and vegetables enriched with AsA, commonly used in animal and human feed.
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Chloroplast ascorbate peroxidases exert an important role in the maintenance of hydrogen peroxide levels in chloroplasts by using ascorbate as the specific electron donor. In this work, we performed a functional study of the stromal APX in rice (OsAPX7) and demonstrated that silencing of OsAPX7 did not impact plant growth, redox state, or photosynthesis parameters. Nevertheless, when subjected to drought stress, silenced plants (APX7i) show a higher capacity to maintain stomata aperture and photosynthesis performance, resulting in a higher tolerance when compared to non-transformed plants. RNA-seq analyses indicate that the silencing of OsAPX7 did not lead to changes in the global expression of genes related to reactive oxygen species metabolism. In addition, the drought-mediated induction of several genes related to the proteasome pathway and the down-regulation of genes related to nitrogen and carotenoid metabolism was impaired in APX7i plants. During drought stress, APX7i showed an up-regulation of genes encoding flavonoid and tyrosine metabolism enzymes and a down-regulation of genes related to phytohormones signal transduction and nicotinate and nicotinamide metabolism. Our results demonstrate that OsAPX7 might be involved in signaling transduction pathways related to drought stress response, contributing to the understanding of the physiological role of chloroplast APX isoforms in rice.
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Ascorbate peroxidases (APXs) are heme peroxidases that remove hydrogen peroxide in different subcellular compartments with concomitant ascorbate cycling. Here, we analysed and discussed phylogenetic and molecular features of the APX family. Ancient APX originated as a soluble stromal enzyme, and early during plant evolution, acquired both chloroplast-targeting and mitochondrion-targeting sequences and an alternative splicing mechanism whereby it could be expressed as a soluble or thylakoid membrane-bound enzyme. Later, independent duplication and neofunctionalization events in some angiosperm groups resulted in individual genes encoding stromal, thylakoidal and mitochondrial isoforms. These data reaffirm the complexity of plant antioxidant defenses that allow diverse plant species to acquire new means to adapt to changing environmental conditions.
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Peroxidases , Tilacoides , Ascorbato Peroxidases/genética , Ascorbato Peroxidases/metabolismo , Tilacoides/metabolismo , Filogenia , Peroxidases/genética , Peroxidases/metabolismo , Cloroplastos/metabolismo , Peróxido de Hidrogênio/metabolismo , Antioxidantes , Regulação da Expressão Gênica de PlantasRESUMO
Latex diagnosis (LD) is applied to optimize the natural rubber production and prevent tapping panel dryness (TPD), a physiological syndrome affecting latex production in Hevea brasiliensis. The reduced thiol content (RSH) is one of the biochemical parameters associated with the risk of TPD. However, RSH is difficult to interpret because of the influence of the environment. In order to better understand the regulation of antioxidants and to better interpret RSH, a key parameter of LD, this study analysed in latex both oxidised and reduced forms of ascorbic acid (AsA) and glutathione, and their cofactors as well as other latex diagnosis parameters in response to harvesting stress (tapping and ethephon stimulation) and TPD occurrence. The content of antioxidants in latex had a high variability among five rubber clones. The concentration in AsA was about ten times higher than GSH in laticifer, GSH accounting for about 50% of RSH. For short-term harvesting stress, RSH increased with tapping frequency and ethephon stimulation. TPD is associated with high latex viscosity and bursting of lysosomal particles called lutoids, as well as for several rubber clones with lower RSH and GSH contents. These results suggest that a high level of RSH shows the capacity of laticifer metabolism to cope with harvesting stress, while a drop in RSH is the sign of long stress related to lower metabolic activity and TPD occurrence. RSH remains an essential physiological parameter to prevent TPD when associated with reference data under low and high harvesting stress. This study paves the way to understand the role of AsA and GSH, and carry out genetic studies of antioxidants.
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Experiments were conducted to investigate whether supplementation of cryopreservation medium with ascorbate, dithiothreitol (DTT) or an inhibitor of caspase-3 (z-DEVD-fmk) could improve post-thaw survival of bovine embryos produced in vitro (IVP). For all experiments, embryos were harvested on day 7 after insemination and subjected to controlled-rate freezing in medium containing 1.5 M ethylene glycol and treatments as described below. In experiments 1-3, embryos were cryopreserved in freezing medium with ascorbate (0, 0.1, 0.3 or 0.5 mM), DTT (0, 50, 100 or 200 µM) and z-DEVD-fmk (0, 50, 100 or 200 µM), respectively. Post-thaw survival was assessed at 24, 48 and 72 h. For experiments 4-5, embryos were cryopreserved in freezing medium with or without 0.1 mM ascorbate. At 24 h post-thaw, embryo total cell number, DNA fragmentation and levels of reactive oxygen species (ROS) were evaluated. Embryos subjected to freezing and thawing in medium supplemented with 0.1 mM ascorbate had greater (p < .05) re-expansion rates at 24, 48 and 72 h and hatching rate at 72 h as compared to embryos not treated with ascorbate. Post-thaw cryosurvival was not affected by the addition of either DTT or z-DEVD-fmk to medium used for cryopreservation. Embryos cryopreserved in medium supplemented with 0.1 mM ascorbate had reduced (p < .001) levels of intracellular ROS and fewer (p < .001) cells with DNA fragmentation. In conclusion, post-thaw survival of bovine IVP embryos is enhanced by supplementation of freezing medium with ascorbate.
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Criopreservação , Embrião de Mamíferos , Animais , Caspase 3 , Inibidores de Caspase , Bovinos , Criopreservação/veterinária , Ditiotreitol/farmacologia , Fertilização in vitro/veterinária , Espécies Reativas de OxigênioRESUMO
Cancer is a disease of high mortality, and its prevalence has increased steadily in the last few years. However, during the last decade, the development of modern chemotherapy schemes, new radiotherapy techniques, targeted therapies and immunotherapy has brought new hope in the treatment of these diseases. Unfortunately, cancer therapies are also associated with frequent and, sometimes, severe adverse events. Ascorbate (ascorbic acid or vitamin C) is a potent water-soluble antioxidant that is produced in most mammals but is not synthesised endogenously in humans, which lack enzymes for its synthesis. Ascorbate has antioxidant effects that correspond closely to the dose administered. Interestingly, this natural antioxidant induces oxidative stress when given intravenously at a high dose, a paradoxical effect due to its interactions with iron. Importantly, this deleterious property of ascorbate can result in increased cell death. Although, historically, ascorbate has been reported to exhibit anti-tumour properties, this effect has been questioned due to the lack of available mechanistic detail. Recently, new evidence has emerged implicating ferroptosis in several types of oxidative stress-mediated cell death, such as those associated with ischemia-reperfusion. This effect could be positively modulated by the interaction of iron and high ascorbate dosing, particularly in cell systems having a high mitotic index. In addition, it has been reported that ascorbate may behave as an adjuvant of favourable anti-tumour effects in cancer therapies such as radiotherapy, radio-chemotherapy, chemotherapy, immunotherapy, or even in monotherapy, as it facilitates tumour cell death through the generation of reactive oxygen species and ferroptosis. In this review, we provide evidence supporting the view that ascorbate should be revisited to develop novel, safe strategies in the treatment of cancer to achieve their application in human medicine.
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Antioxidantes , Neoplasias , Animais , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Ácido Ascórbico/farmacologia , Ácido Ascórbico/uso terapêutico , Humanos , Ferro/metabolismo , Mamíferos/metabolismo , Neoplasias/tratamento farmacológico , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismoRESUMO
Increased synthesis of H2O2 is observed during the initiation of fruit ripening. However, its association with plant cell processes triggering the maturation of fruit has not yet been demonstrated. The aim of this work is to investigate whether H2O2 participates in the tomato ripening process and particularly through its association with the ethylene signaling pathway. The experiments were carried out with two ethyl methanesulfonate mutant lines of Micro-Tom tomato deficient in GDP-L-galactose phosphorylase activity and displaying lower ascorbic acid content than the corresponding parental genotype (i.e. wild type). Plants were subjected to a high irradiance (HI) treatment to stimulate H2O2 synthesis. HI treatment enhanced H2O2 production and reduced the timing of fruit ripening in both mutants and wild-type fruits. These results could be linked to an increase of the expression of H2O2-related genes and changes in the expression of ethylene-related genes. The fruit H2O2 production increased or decreased after applying the treatments that induced ethylene synthesis or blocked its action, respectively. The results presented in this work give an evidence of the association of redox and hormonal components during fruit ripening in which H2O2 participates downstream in the events regulated by ethylene.
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Solanum lycopersicum , Etilenos/metabolismo , Frutas/genética , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Peróxido de Hidrogênio/metabolismo , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Salicylic acid (SA) has been shown to ameliorate drought stress. However, physiological and biochemical mechanisms involved in drought stress tolerance induced by SA in plants have not been well understood. Thus, this study aimed to study the role of SA application on enzymatic and non-enzymatic antioxidants, photosynthetic performance, and plant growth in A. chilensis plants subjected to moderate drought stress. One-year-old A. chilensis plants were subjected to 100% and 60% of field capacity. When plants reached moderate drought stress (average of stem water potential of -1.0 MPa, considered as moderate drought stress), a single SA application was performed on plants. Then, physiological and biochemical features were determined at different times during 14 days. Our study showed that SA application increased 13.5% plant growth and recovered 41.9% AN and 40.7% gs in drought-stressed plants on day 3 compared to drought-stressed plants without SA application. Interestingly, SOD and APX activities were increased 85% and 60%, respectively, in drought-stressed SA-treated plants on day 3. Likewise, SA improved 30% total phenolic content and 60% antioxidant capacity in drought-stressed A. chilensis plants. Our study provides insight into the SA mechanism to tolerate moderate drought stress in A. chilensis plants.
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Gracilariales is a clade of florideophycean red macroalgae known for being the main source of agar. We present a de novo genome assembly and annotation of Gracilaria domingensis, an agarophyte alga with flattened thallus widely distributed along Central and South American Atlantic intertidal zones. In addition to structural analysis, an organizational comparison was done with other Rhodophyta genomes. The nuclear genome has 78 Mbp, with 11,437 predicted coding genes, 4,075 of which did not have hits in sequence databases. We also predicted 1,567 noncoding RNAs, distributed in 14 classes. The plastid and mitochondrion genome structures were also obtained. Genes related to agar synthesis were identified. Genes for type II galactose sulfurylases could not be found. Genes related to ascorbate synthesis were found. These results suggest an intricate connection of cell wall polysaccharide synthesis and the redox systems through the use of L-galactose in Rhodophyta. The genome of G. domingensis should be valuable to phycological and aquacultural research, as it is the first tropical and Western Atlantic red macroalgal genome to be sequenced.
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Genoma Mitocondrial , Gracilaria , Rodófitas , Ágar/metabolismo , Galactose/metabolismo , Gracilaria/genética , Rodófitas/genética , Rodófitas/metabolismoRESUMO
Ascorbate peroxidase (APX), Monodehydroascorbate Reductase (MDAR), Dehydroascorbate Reductase (DHAR) and Glutathione Reductase (GR) enzymes participate in the ascorbate-glutathione cycle, which exerts a central role in the antioxidant metabolism in plants. Despite the importance of this antioxidant system in different signal transduction networks related to development and response to environmental stresses, the pathway has not yet been comprehensively characterized in many crop plants. Among different eudicotyledons, the Euphorbiaceae family is particularly diverse with some species highly tolerant to drought. Here the APX, MDAR, DHAR, and GR genes in Ricinus communis, Jatropha curcas, Manihot esculenta, and Hevea brasiliensis were identified and characterized. The comprehensive phylogenetic and genomic analyses allowed the classification of the genes into different classes, equivalent to cytosolic, peroxisomal, chloroplastic, and mitochondrial enzymes, and revealed the duplication events that contribute to the expansion of these families within plant genomes. Due to the high drought stress tolerance of Ricinus communis, the expression patterns of ascorbate-glutathione cycle genes in response to drought were also analyzed in leaves and roots, indicating a differential expression during the stress. Altogether, these data contributed to the characterization of the expression pattern and evolutionary analysis of these genes, filling the gap in the proposed functions of core components of the antioxidant mechanism during stress response in an economically relevant group of plants.
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Abstract: Boron is one of the most important micronutrients for plants. Plants may suffer from deficiency or with boron toxicity. Boron plays a role in significant physiological and biochemical events in plants such as synthesis of the cell wall, membrane integrity, antioxidation, transport of photosynthesis products to other organs of the plant. The enzyme activities of ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR) and superoxide dismutase (SOD) in three different safflower cultivars (Balcı, Dinçer and Remzibey) subjected to different boric acid concentrations (0, 5, 10, 15 mM) were measured spectrophotometrically, and the changes in the expression levels of the genes that encode these enzymes were obtained by quantitative RT-qPCR. When both the spectrophotometric measurements and the mRNA values were evaluated together, both the activity and mRNA values of APX and GR enzymes were found to be the highest in the Dinçer cultivar among the varieties treated with 15 mM boric acid, while the lowest values of these enzymes were determined in the Remzibey cultivar. According to the RT-qPCR results, the lowest SOD and CAT values were determined in Remzibey. The Dinçer cultivar was found to have the highest antioxidant capacity (APX, GR) to cope with oxidative stress caused by boric acid application at high concentrations. The sensitive Remzibey cultivar was found to have the lowest antioxidant capacity to cope with such oxidative stress. Balcı was found to be closer to Dinçer than to Remzibey in terms of boron tolerance. As a result, the boron-sensitive cultivar had low antioxidant activity.
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Oligoelementos/administração & dosagem , Boro/administração & dosagem , Produção Agrícola , Carthamus tinctorius/metabolismo , Antioxidantes/metabolismo , Oligoelementos/toxicidade , Boro/toxicidade , Expressão Gênica/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Carthamus tinctorius/enzimologia , Carthamus tinctorius/genéticaRESUMO
The black oat (Avena strigosa Schreb.) stands out as a forage of great importance in Brazilian agriculture. However, the productivity and quality of this forage can be affected by abiotic factors, such as temperature and water availability, which affect the physiological processes and facilitate the accumulation of free radicals (reactive oxygen species - ROS). Thus, the objective of this study was to understand the biochemical changes in black oat plants subjected to water deficit at different temperatures. Experiments were conducted in a greenhouse in two experimental periods, which presented an average temperature of 20 °C and 24 °C, respectively. Black oat seeds, of the variety IAPAR 61, were sown in pots and the plants were irrigated for 60 days. After which, the pots were covered with plastic bags and the irrigation was suspended. The analyses were carried out in five periods of evaluation - M1: plants before the suspensionof irrigation, M2: plants at the first wilting point, M3: three days after plastic removal and irrigation return, M4: four days after M3 and before the second suspension of irrigation, and M5: the second wilting point. The levels of total protein and malondialdehyde (MDA), and the activity of the enzymes catalase (CAT) and ascorbate peroxidase (APX), were analyzed. The experimental design was completely randomized, with six replications, in a factorial scheme of average temperature × water management × periods of evaluation, and the means were compared by Tukey's test at 5%. In response to water deficiency and temperature increase, black oat plants increased their levels of total soluble proteins, and there was greater lipid peroxidation due to the increase in malondialdehyde content. There was no change in the activity of the enzymes catalase and ascorbate peroxidase under water deficit, and these activities decreased with increasing temperature.
A aveia-preta (Avena strigosa Schreb.) destaca-se como uma forragem de grande importância na agricultura brasileira. Porém, a produtividade e a qualidade dessa forragem podem ser afetadas por fatores abióticos, como temperatura e disponibilidade de água, que afetam os processos fisiológicos e facilitam o acúmulo de radicais livres (espécies reativas de oxigênio - ROS). Assim, o objetivo deste estudo foi compreender as alterações bioquímicas em plantas de aveia preta submetidas ao déficit hídrico em diferentes temperaturas. Os experimentos foram conduzidos em casa de vegetação em dois períodos experimentais, os quais apresentaram temperatura média de 20 °C e 24 °C, respectivamente. Sementes de aveia preta, variedade IAPAR 61, foram semeadas em vasos e as plantas irrigadas por 60 dias. Depois disso, os vasos foram cobertos com sacos plásticos e a irrigação foi suspensa. As análises foram realizadas em cinco períodos de avaliação - M1: plantas antes da suspensão da irrigação, M2: plantas no primeiro ponto de murcha, M3: três dias após a retirada do plástico e retorno da irrigação, M4: quatro dias após M3 e antes do segundo suspensão da irrigação e M5: o segundo ponto de murcha. Foram analisados os níveis de proteína total e malondialdeído (MDA) e a atividade das enzimas catalase (CAT) e ascorbato peroxidase (APX). O delineamento experimental foi inteiramente casualizado, com seis repetições, em esquema fatorial temperatura média × manejo da água × períodos de avaliação, e as médias foram comparadas pelo teste de Tukey a 5%. Em resposta à deficiência hídrica e ao aumento da temperatura, as plantas de aveia preta aumentaram seus níveis de proteínas solúveis totais e houve maior peroxidação lipídica devido ao aumento do teor de malondialdeído. Não houve alteração na atividade das enzimas catalase e ascorbato peroxidase sob déficit hídrico, sendo que essas atividades diminuíram com o aumento da temperatura.
Assuntos
Avena/metabolismo , Desidratação/diagnóstico , GeumRESUMO
The black oat (Avena strigosa Schreb.) stands out as a forage of great importance in Brazilian agriculture. However, the productivity and quality of this forage can be affected by abiotic factors, such as temperature and water availability, which affect the physiological processes and facilitate the accumulation of free radicals (reactive oxygen species - ROS). Thus, the objective of this study was to understand the biochemical changes in black oat plants subjected to water deficit at different temperatures. Experiments were conducted in a greenhouse in two experimental periods, which presented an average temperature of 20 °C and 24 °C, respectively. Black oat seeds, of the variety IAPAR 61, were sown in pots and the plants were irrigated for 60 days. After which, the pots were covered with plastic bags and the irrigation was suspended. The analyses were carried out in five periods of evaluation - M1: plants before the suspensionof irrigation, M2: plants at the first wilting point, M3: three days after plastic removal and irrigation return, M4: four days after M3 and before the second suspension of irrigation, and M5: the second wilting point. The levels of total protein and malondialdehyde (MDA), and the activity of the enzymes catalase (CAT) and ascorbate peroxidase (APX), were analyzed. The experimental design was completely randomized, with six replications, in a factorial scheme of average temperature × water management × periods of evaluation, and the means were compared by Tukey's test at 5%. In response to water deficiency and temperature increase, black oat plants increased their levels of total soluble proteins, and there was greater lipid peroxidation due to the increase in malondialdehyde content. There was no change in the activity of the enzymes catalase and ascorbate peroxidase under water deficit, and these activities decreased with increasing temperature.(AU)
A aveia-preta (Avena strigosa Schreb.) destaca-se como uma forragem de grande importância na agricultura brasileira. Porém, a produtividade e a qualidade dessa forragem podem ser afetadas por fatores abióticos, como temperatura e disponibilidade de água, que afetam os processos fisiológicos e facilitam o acúmulo de radicais livres (espécies reativas de oxigênio - ROS). Assim, o objetivo deste estudo foi compreender as alterações bioquímicas em plantas de aveia preta submetidas ao déficit hídrico em diferentes temperaturas. Os experimentos foram conduzidos em casa de vegetação em dois períodos experimentais, os quais apresentaram temperatura média de 20 °C e 24 °C, respectivamente. Sementes de aveia preta, variedade IAPAR 61, foram semeadas em vasos e as plantas irrigadas por 60 dias. Depois disso, os vasos foram cobertos com sacos plásticos e a irrigação foi suspensa. As análises foram realizadas em cinco períodos de avaliação - M1: plantas antes da suspensão da irrigação, M2: plantas no primeiro ponto de murcha, M3: três dias após a retirada do plástico e retorno da irrigação, M4: quatro dias após M3 e antes do segundo suspensão da irrigação e M5: o segundo ponto de murcha. Foram analisados os níveis de proteína total e malondialdeído (MDA) e a atividade das enzimas catalase (CAT) e ascorbato peroxidase (APX). O delineamento experimental foi inteiramente casualizado, com seis repetições, em esquema fatorial temperatura média × manejo da água × períodos de avaliação, e as médias foram comparadas pelo teste de Tukey a 5%. Em resposta à deficiência hídrica e ao aumento da temperatura, as plantas de aveia preta aumentaram seus níveis de proteínas solúveis totais e houve maior peroxidação lipídica devido ao aumento do teor de malondialdeído. Não houve alteração na atividade das enzimas catalase e ascorbato peroxidase sob déficit hídrico, sendo que essas atividades diminuíram com o aumento da temperatura.(AU)