Biochar and Chitosan Regulate Antioxidant Defense and Methylglyoxal Detoxification Systems and Enhance Salt Tolerance in Jute (Corchorus olitorius L.)

We investigated the role of biochar and chitosan in mitigating salt stress in jute (Corchorus olitorius L. cv. O-9897) by exposing twenty-day-old seedlings to three doses of salt (50, 100, and 150 mM NaCl). Biochar was pre-mixed with the soil at 2.0 g kg−1 soil, and chitosan-100 was applied through irrigation at 100 mg L−1. Exposure to salt stress notably increased lipid peroxidation, hydrogen peroxide content, superoxide radical levels, electrolyte leakage, lipoxygenase activity, and methylglyoxal content, indicating oxidative damage in the jute plants. Consequently, the salt-stressed plants showed reduced growth, biomass accumulation, and disrupted water balance. A profound increase in proline content was observed in response to salt stress. Biochar and chitosan supplementation significantly mitigated the deleterious effects of salt stress in jute by stimulating both non-enzymatic (e.g., ascorbate and glutathione) and enzymatic (e.g., ascorbate peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase, glutathione reductase superoxide dismutase, catalase, peroxidase, glutathione S-transferase, glutathione peroxidase) antioxidant systems and enhancing glyoxalase enzyme activities (glyoxalase I and glyoxalase II) to ameliorate reactive oxygen species damage and methylglyoxal toxicity, respectively. Biochar and chitosan supplementation increased oxidative stress tolerance and improved the growth and physiology of salt-affected jute plants, while also significantly reducing Na+ accumulation and ionic toxicity and decreasing the Na+/K+ ratio. These findings support a protective role of biochar and chitosan against salt-induced damage in jute plants.

Suppression of the Lycopene Cyclase Gene Causes Downregulation of Ascorbate Peroxidase Activity and Decreased Glutathione Pool Size, Leading to H2O2 Accumulation in Euglena gracilis

Carotenoids are photosynthetic pigments and hydrophobic antioxidants that are necessary for the survival of photosynthetic organisms, including the microalga Euglena gracilis. In the present study, we identified an uncharacterized gene encoding the E. gracilis β-carotene synthetic enzyme lycopene cyclase (EgLCY) and discovered a relationship between EgLCY-mediated carotenoid synthesis and the reactive oxygen species (ROS) scavenging system ascorbate-glutathione cycle. The EgLCY cDNA sequence was obtained via homology searching E. gracilis transcriptome data. An enzyme assay using Escherichia coli demonstrated that EgLCY converts lycopene to β-carotene. E. gracilis treated with EgLCY double-stranded RNA (dsRNA) produced colorless cells with hypertrophic appearance, inhibited growth, and marked decrease in carotenoid and chlorophyll content, suggesting that EgLCY is essential for the synthesis of β-carotene and downstream carotenoids, which are abundant and physiologically functional. In EgLCY dsRNA-treated cells, the ascorbate-glutathione cycle, composed of ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDAR), and glutathione reductase (GR), was unusually modulated; APX and GR activities significantly decreased, whereas DHAR and MDAR activities increased. Ascorbate content was significantly increased and glutathione content significantly decreased in EgLCY dsRNA-treated cells and was correlated with their recycling enzyme activities. Fluorescent imaging demonstrated that EgLCY dsRNA-treated cells accumulated higher levels of H2O2 compared to wild-type cells. Taken together, this study revealed that EgLCY-mediated synthesis of β-carotene and downstream carotenoid species upregulates APX activity and increases glutathione pool size for H2O2 scavenging. Our study suggests a possible relationship between carotenoid synthesis and the ascorbate-glutathione cycle for ROS scavenging in E. gracilis.

Metabolic Changes in Seed Embryos of Hypoxia-Tolerant Rice and Hypoxia-Sensitive Barley at the Onset of Germination

Rice (Oryza sativa L.) and barley (Hordeum vulgare L.) are the cereal species differing in tolerance to oxygen deficiency. To understand metabolic differences determining the sensitivity to low oxygen, we germinated rice and barley seeds and studied changes in the levels of reactive oxygen species (ROS) and reactive nitrogen species (RNS), activities of the enzymes involved in their scavenging, and measured cell damage parameters. The results show that alcohol dehydrogenase activity was higher in rice than in barley embryos providing efficient anaerobic fermentation. Nitric oxide (NO) levels were also higher in rice embryos indicating higher NO turnover. Both fermentation and NO turnover can explain higher ATP/ADP ratio values in rice embryos as compared to barley. Rice embryos were characterized by higher activity of S-nitrosoglutathione reductase than in barley and a higher level of free thiols in proteins. The activities of antioxidant enzymes (superoxide dismutase, ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase) in imbibed embryos were higher in rice than in barley, which corresponded to the reduced levels of ROS, malonic dialdehyde and electrolyte leakage. The observed differences in metabolic changes in embryos of the two cereal species differing in tolerance to hypoxia can partly explain the adaptation of rice to low oxygen environments.

Praseodymium enhanced the tolerance of maize seedlings subjected to cadmium stress by up-regulating the enzymes in the regeneration and biosynthetic pathways of ascorbate and glutathione

To test whether praseodymium (Pr) regulates cadmium (Cd) tolerance, we explored the effects of Pr on enzymatic activities in the regeneration and biosynthetic pathways of ascorbate and glutathione in maize seedlings under Cd stress. The findings demonstrated that Cd stress increased enzymatic activities in the regeneration pathway (ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR)) and in the biosynthetic pathway of ascorbate and glutathione (γ-ECS and GalLDH), as well as ascorbate (AsA) and glutathione (GSH) contents. However, Cd stress significantly decreased AsA/dehydroascorbic acid (DHA) ratio and GSH/oxidised glutathione (GSSG) ratio, net photosynthetic rate (P_n), chlorophylls (Chl) and carotenoids (Car) contents, maximum photochemical efficiency of PSII (F_v/F_m), photochemical quenching (qP) and quantum efficiency of PSII photochemistry (Φ_PSII), as well as plant height and biomass. Application of Pr to Cd-stressed seedlings enhanced above enzymatic activities, AsA and GSH contents, AsA/DHA and GSH/GSSG ratios, P_n, Chl and Car contents, F_v/F_m, qP and Φ_PSII, as well as plant height and biomass. Meanwhile, the application of Pr to Cd-stressed seedlings reduced malondialdehyde (MDA) content and electrolyte leakage. The above results indicated that Pr enhanced Cd tolerance of maize by up-regulating enzymatic activities in regeneration and biosynthetic pathways of ascorbate and glutathione.

Distribution and Functions of Monodehydroascorbate Reductases in Plants: Comprehensive Reverse Genetic Analysis of Arabidopsis thaliana Enzymes

Monodehydroascorbate reductase (MDAR) is an enzyme involved in ascorbate recycling. Arabidopsis thaliana has five MDAR genes that encode two cytosolic, one cytosolic/peroxisomal, one peroxisomal membrane-attached, and one chloroplastic/mitochondrial isoform. In contrast, tomato plants possess only three enzymes, lacking the cytosol-specific enzymes. Thus, the number and distribution of MDAR isoforms differ according to plant species. Moreover, the physiological significance of MDARs remains poorly understood. In this study, we classify plant MDARs into three classes: class I, chloroplastic/mitochondrial enzymes; class II, peroxisomal membrane-attached enzymes; and class III, cytosolic/peroxisomal enzymes. The cytosol-specific isoforms form a subclass of class III and are conserved only in Brassicaceae plants. With some exceptions, all land plants and a charophyte algae, Klebsormidium flaccidum, contain all three classes. Using reverse genetic analysis of Arabidopsis thaliana mutants lacking one or more isoforms, we provide new insight into the roles of MDARs; for example, (1) the lack of two isoforms in a specific combination results in lethality, and (2) the role of MDARs in ascorbate redox regulation in leaves can be largely compensated by other systems. Based on these findings, we discuss the distribution and function of MDAR isoforms in land plants and their cooperation with other recycling systems.

Response to Antimony Toxicity in Dittrichia viscosa Plants: ROS, NO, H2S, and the Antioxidant System

Dittrichia viscosa plants were grown hydroponically with different concentrations of Sb. There was preferential accumulation of Sb in roots. Fe and Cu decreased, while Mn decreased in roots but not in leaves. Chlorophyll content declined, but the carotenoid content increased, and photosynthetic efficiency was unaltered. O2●− generation increased slightly, while lipid peroxidation increased only in roots. H2O2, NO, ONOO−, S-nitrosothiols, and H2S showed significant increases, and the enzymatic antioxidant system was altered. In roots, superoxide dismutase (SOD) and monodehydroascorbate reductase (MDAR) activities declined, dehydroscorbate reductase (DHAR) rose, and ascorbate peroxidase (APX), peroxidase (POX), and glutathione reductase (GR) were unaffected. In leaves, SOD and POX increased, MDAR decreased, and APX was unaltered, while GR increased. S-nitrosoglutathione reductase (GSNOR) and l-cysteine desulfhydrilase (l-DES) increased in activity, while glutathione S-transferase (GST) decreased in leaves but was enhanced in roots. Components of the AsA/GSH cycle decreased. The great capacity of Dittrichia roots to accumulate Sb is the reason for the differing behaviour observed in the enzymatic antioxidant systems of the two organs. Sb appears to act by binding to thiol groups, which can alter free GSH content and SOD and GST activities. The coniferyl alcohol peroxidase activity increased, possibly to lignify the roots’ cell walls. Sb altered the ROS balance, especially with respect to H2O2. This led to an increase in NO and H2S acting on the antioxidant system to limit that Sb-induced redox imbalance. The interaction NO, H2S and H2O2 appears key to the response to stress induced by Sb. The interaction between ROS, NO, and H2S appears to be involved in the response to Sb.

Supplemental Selenium and Boron Mitigate Salt-Induced Oxidative Damages in Glycine max L.

The present investigation was executed with an aim to evaluate the role of exogenous selenium (Se) and boron (B) in mitigating different levels of salt stress by enhancing the reactive oxygen species (ROS) scavenging, antioxidant defense and glyoxalase systems in soybean. Plants were treated with 0, 150, 300 and 450 mM NaCl at 20 days after sowing (DAS). Foliar application of Se (50 µM Na2SeO4) and B (1 mM H3BO3) was accomplished individually and in combined (Se+B) at three-day intervals, at 16, 20, 24 and 28 DAS under non-saline and saline conditions. Salt stress adversely affected the growth parameters. In salt-treated plants, proline content and oxidative stress indicators such as malondialdehyde (MDA) content and hydrogen peroxide (H2O2) content were increased with the increment of salt concentration but the relative water content decreased. Due to salt stress catalase (CAT), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glyoxalase I (Gly I) and glyoxalase II (Gly II) activity decreased. However, the activity of ascorbate peroxidase (APX), glutathione reductase (GR), glutathione peroxidase (GPX), glutathione S-transferase (GST) and peroxidase (POD) increased under salt stress. On the contrary, supplementation of Se, B and Se+B enhanced the activities of APX, MDHAR, DHAR, GR, CAT, GPX, GST, POD, Gly I and Gly II which consequently diminished the H2O2 content and MDA content under salt stress, and also improved the growth parameters. The results reflected that exogenous Se, B and Se+B enhanced the enzymatic activity of the antioxidant defense system as well as the glyoxalase systems under different levels of salt stress, ultimately alleviated the salt-induced oxidative stress, among them Se+B was more effective than a single treatment.

Phytochemical Analysis Reveals an Antioxidant Defense Response in Lonicera Japonica To Cadmium-Induced Oxidative Stress

Cadmium (Cd), though potentially beneficial at lower levels to some plant species, at higher levels is a toxic metal that is detrimental to plant growth and development. Cadmium is also a carcinogen to humans and other contaminated plant consumers, affecting the kidneys and reducing bone strength. In this study we investigated responses of growth, chlorophyll content, reactive oxygen species (ROS) levels, and antioxidant responses to Cd2+ in honeysuckle leaves (Lonicera japonica Thunb.), a potential cadmium hyperaccumulator. Results indicated that plant height, dry weight, leaf area, and chlorophyll content increased when honeysuckle was exposed to 10 or 30 mg/kg Cd2+ (low concentration). However, in response to 150 or 200 mg/kg Cd2+ (high concentration) these growth parameters and chlorophyll content significantly decreased relative to untreated control plant groups. Higher levels of superoxide radical (O2.−) and hydrogen peroxide (H2O2) were observed in high concentration Cd2+ groups. The activities of ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR), and glutathione reductase (GR) were enhanced with exposure to increasing levels of Cd2+. Additionally, the AsA-GSH cycle was activated for the removal of H2O2 in honeysuckle in response to elevated Cd2+. The Pearson correlation analysis, a redundancy analysis (RDA), and a permutation test indicated that proline and APX were dominant antioxidants for removing O2.− and H2O2. The antioxidants GSH and NPTs also increased as the concentration of Cd2+ increased.

Maintenance of Postharvest Quality and Reactive Oxygen Species Homeostasis of Pitaya Fruit by Essential Oil p-Anisaldehyde Treatment

The performance of p-Anisaldehyde (PAA) for preserving pitaya fruit quality and the underpinning regulatory mechanism were investigated in this study. Results showed that PAA treatment significantly reduced fruit decay, weight loss and loss of firmness, and maintained higher content of total soluble solids, betacyanins, betaxanthins, total phenolics and flavonoids in postharvest pitaya fruits. Compared with control, the increase in hydrogen peroxide (H2O2) content and superoxide anion (O2•−) production was inhibited in fruit treated with PAA. Meanwhile, PAA significantly improved the activity of antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT). Moreover, PAA-treated pitaya fruit maintained higher ascorbic acid (AsA) and reduced-glutathione (GSH) content but lower dehydroascorbate (DHA) and oxidized glutathione (GSSG) content, thus sustaining higher ratio of AsA/DHA and GSH/GSSG. In addition, activities of ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydrogenation ascorbic acid reductase (DHAR), as well as the expression of HpSOD, HpPOD, HpCAT, HpAPX, HpGR, HpDHAR and HpMDHAR, were enhanced after PAA treatment. The findings suggest that postharvest application of PAA may be a reliable method to control postharvest decay and preserve quality of harvested pitaya fruit by enhancing the antioxidant potential of the AsA-GSH cycle and activating an antioxidant defense system to alleviate reactive oxygen species (ROS) accumulation.