水分胁迫下玉米叶片光合的活性氧限制

玉米植株叶片在水分胁迫下,活性氧代谢平衡失调。随着胁迫强度的增加,O^-₂和H₂O₂大量产生,导致叶绿素氧化破坏。光合受抑在中度至重度胁迫时,非气孔因素中活性氧伤害起主导作用。抗旱杂交种抵御活性氧伤害的能力强于干旱敏感杂交种。     

An Ethanol Extract Derived from Bonnemaisonia hamifera Scavenges Ultraviolet B (UVB) Radiation-Induced Reactive Oxygen Species and Attenuates UVB-Induced Cell Damage in Human Keratinocytes

The present study investigated the photoprotective properties of an ethanol extract derived from the red alga Bonnemaisonia hamifera against ultraviolet B (UVB)-induced cell damage in human HaCaT keratinocytes. The Bonnemaisonia hamifera ethanol extract (BHE) scavenged the superoxide anion generated by the xanthine/xanthine oxidase system and the hydroxyl radical generated by the Fenton reaction (FeSO₄ + H₂O₂), both of which were detected by using electron spin resonance spectrometry. In addition, BHE exhibited scavenging activity against the 1,1-diphenyl-2-picrylhydrazyl radical and intracellular reactive oxygen species (ROS) that were induced by either hydrogen peroxide or UVB radiation. BHE reduced UVB-induced apoptosis, as shown by decreased apoptotic body formation and DNA fragmentation. BHE also attenuated DNA damage and the elevated levels of 8-isoprostane and protein carbonyls resulting from UVB-mediated oxidative stress. Furthermore, BHE absorbed electromagnetic radiation in the UVB range (280–320 nm). These results suggest that BHE protects human HaCaT keratinocytes against UVB-induced oxidative damage by scavenging ROS and absorbing UVB photons, thereby reducing injury to cellular components.     

Anti-Aging Effects of R-Phycocyanin from Porphyra haitanensis on HUVEC Cells and Drosophila melanogaster

Aging has become a global public health challenge. Many studies have revealed that the excessive generation of ROS and oxidative stress could be the major causative factors contributing to aging. In this study, R-phycocyanin (R-PC) was isolated from Porphyra haitanensis, and its anti-aging ability was explored by natural aging Drosophila melanogaster and H₂O₂-induced HUVEC cells as the aging model. Results showed that R-PC α and β subunits expressed have antioxidant activity and can inhibit the generation of radicals, exhibiting a protective effect against H₂O₂-induced apoptotic HUVEC cells death. R-PC prevented the H₂O₂-induced HUVEC cell cycle phase arrest by regulating cell cycle-related protein. Furthermore, R-PC prevented the H₂O₂-induced HUVEC cell cycle phase arrest by regulating cell-cycle-related protein expression. In vivo study also indicated that R-PC significantly increased the survival time and alleviated the oxidative stress of Drosophila melanogaster. Moreover, R-PC notably decreased levels of ROS in natural aging flies and inhibited lipid peroxidation by enhancing the expressions of the endogenous stress marker genes (SOD1, SOD2, CAT of Drosophila melanogaster). Taken together, a study on the antioxidation extract from Porphyra haitanensis, such as R-PC, may open a new window for the prevention of anti-aging.     

Induction of Freezing Tolerance by the Application of Hydrogen Peroxide and Salicylic Acid as Tuber-Dip or Canopy Spraying in <Emphasis Type="Italic">Solanum tuberosum</Emphasis> L. Plants

Low temperature stress is a current challenge to plants that is associated with climate change. In plants, exposure to extreme temperatures is followed by the accumulation of reactive oxygen species, such as hydrogen peroxide (H₂O₂), leading to oxidative stress. Salicylic acid (SA) and H₂O₂ mediate the tolerance responses to stress and have been reported to induce freezing tolerance in potato microplants. The objectives of the present investigation were (1) to evaluate the short- and long-term effects of H₂O₂ and SA treatments on freezing tolerance in potato (Solanum tuberosum L.) plants grown from tubers and (2) to analyse the relationship between catalase (CAT) activity and H₂O₂ concentration associated with freezing tolerance responses. We observed the lowest freezing survival rates in 45-day-old potato plants (cv. Granate) compared to younger plants. The two treatments consisted of (1) the tuber-dip (long-term) treatment in which sprouted minitubers were saturated for 1 h in SA 10^?5 M or H₂O₂ 1 mM and planted in soil under greenhouse conditions and (2) the crop-spray (short-term) treatment in which plants 5–8 cm high were sprayed twice a week with SA 10^?5 M or H₂O₂ 1 mM until 45 days of age. In all treatments, 45-day-old plants were then exposed to ? 6?±?1 °C for 4 h. The survival rate was measured 15 days after freezing. CAT and H₂O₂ measurements were performed 1 h before and after the freezing treatment. The results showed that SA and H₂O₂ induced freezing tolerance in both the short- and long-term treatments. Survival was significantly higher in SA- and H₂O₂-treated plants than in control plants. In both the long- and short-term treatments this higher survival was associated with lower internal H₂O₂ concentrations after freezing compared with control plants and decreasing oxidative stress. SA and H₂O₂ induced different levels of CAT activity after freezing compared to that found in the control plants in the long- and the short-term treatments. These results suggest the SA and H₂O₂ function in independent pathways in terms of their induction of freezing tolerance that depends on the method the treatment was applied, by spraying the canopy or by immersion of the sprouted seed tuber.     

Bioprospecting of Less-Polar Constituents from Endemic Brown Macroalga Fucus virsoides J. Agardh from the Adriatic Sea and Targeted Antioxidant Effects In Vitro and In Vivo (Zebrafish Model)

The endemic brown macroalga Fucus virsoides J. Agardh from the Adriatic Sea was in the focus of the present research. The volatiles of fresh (FrFv) and air-dried (DrFv) samples of F. virsoides obtained by headspace solid-phase microextraction (HS-SPME) and hydrodistillation (HD) were analyzed by gas chromatography equipped with flame ionization detector and mass spectrometry (GC-FID/MS). The major HS-FrFv compound was pentadecane (61.90–71.55%) followed by pentadec-1-ene (11.00–7.98%). In HS-DrFv, pentadec-1-ene was not present, and few lower aliphatic compounds appeared, as well as benzaldehyde and benzyl alcohol. In HD-FrFv, particularly abundant were alkenes (such as pentadec-1-ene (19.32%), or (E)-pentadec-7-ene (8.35%)). In HD-DrFv, more oxidation products were present (e.g., carbonyl compounds such as tridecanal (18.51%)). The fatty acids profile of freeze-dried sample (FdFv) after conversion to methyl esters was determined by GC-FID, and oleic acid was dominant (42.28%), followed by arachidonic acid (15.00%). High-performance liquid chromatography-high-resolution mass spectrometry with electrospray ionization (HPLC-ESI-HRMS) was used for the screening of less polar fractions (F3 and F4) of F. virsoides. Mono- and diglycerides of stearic, palmitic, oleic, and arachidonic acids were found. Terpenoids and steroids comprised the compounds C₂₀H₃₀₍₃₂₎O₂ and C₂₉H₄₈O₍₂₎. Among carotenoids, fucoxanthin was identified. Chlorophyll derivatives were also found (C₅₅H₇₄₍₇₂₎N₄O_(5-7)), dominated by pheophytin a. The antioxidant activity of the fractions was investigated by in vitro assays (oxygen radical absorbance capacity (ORAC), reduction of radical cation (ABTS•⁺), 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay, and ferric reducing antioxidant power (FRAP)) and by in vivo zebrafish model (along with fish embryotoxicity). In vitro experiments proved good radical scavenging abilities of F3 and F4 fractions, which were additionally supported by the protective effect against hydrogen peroxide-induced oxidative stress in zebrafish embryos.     

Evaluation of the Effects of Fucoidans from Fucus Species and Laminaria hyperborea against Oxidative Stress and Iron-Dependent Cell Death

Fucoidans are algal polysaccharides that exhibit protective properties against oxidative stress. The aim of this study was to investigate different fucoidans from brown seaweeds for their ability to protect against iron-dependent oxidative stress (ferroptosis), a main hallmark of retinal and brain diseases, including hemorrhage. We investigated five new high-molecular weight fucoidan extracts from Fucus vesiculosus, F. serratus, and F. distichus subsp. evanescens, a previously published Laminaria hyperborean extract, and commercially available extracts from F. vesiculosus and Undaria pinnatifida. We induced oxidative stress by glutathione depletion (erastin) and H₂O₂ in four retinal and neuronal cell lines as well as primary cortical neurons. Only extracts from F. serratus, F. distichus subsp. evanescens, and Laminaria hyperborea were partially protective against erastin-induced cell death in ARPE-19 and OMM-1 cells, while none of the extracts showed beneficial effects in neuronal cells. Protective fucoidans also attenuated the decrease in protein levels of the antioxidant enzyme GPX4, a key regulator of ferroptosis. This comprehensive analysis demonstrates that the antioxidant abilities of fucoidans may be cell type-specific, besides depending on the algal species and extraction method. Future studies are needed to further characterize the health-benefiting effects of fucoidans and to determine the exact mechanism underlying their antioxidative abilities.     

Identifying Potential Antioxidant Properties from the Viscera of Sea Snails (Turbo cornutus)

Turbo cornutus, the horned turban sea snail, is found along the intertidal and basaltic shorelines of Jeju Island, Korea. T. cornutus feeds on seaweeds (e.g., Undaria sp., and Ecklonia sp.) composed of diverse antioxidants. This study identified potential antioxidant properties from T. cornutus viscera tissues. Diverse extracts were evaluated for their hydrogen peroxide (H₂O₂) scavenging activities. T. cornutus viscera protamex-assisted extracts (TVP) were purified by gel filtration chromatography (GFC), and potential antioxidant properties were analyzed for their amino acid sequences and its peroxidase inhibition effects by in silico molecular docking and in vitro analysis. According to the results, T. cornutus viscera tissues are composed of many protein contents with each over 50%. Among the extracts, TVP possessed the highest H₂O₂ scavenging activity. In addition, TVP-GFC-3 significantly decreased intracellular reactive oxygen species (ROS) levels and increased cell viability in H₂O₂-treated HepG2 cells without cytotoxicity. TVP-GFC-3 comprises nine low molecular bioactive peptides (ELR, VGPQ, TDY, ALPHA, PAH, VDY, WSDK, VFSP, and FAPQY). Notably, the peptides dock to the active site of the myeloperoxidase (MPO), especially TDY and FAPQY showed the MPO inhibition effects with IC₅₀ values of 646.0 ± 45.0 µM and 57.1 ± 17.7 µM, respectively. Altogether, our findings demonstrated that T. cornutus viscera have potential antioxidant properties that can be used as high value-added ingredients.     

Antioxidant Activity of Gracilaria lemaneiformis Polysaccharide Degradation Based on Nrf-2/Keap-1 Signaling Pathway in HepG2 Cells with Oxidative Stress Induced by H2O2

The objective of this research was to investigate the antioxidant activity of Gracilaria lemaneiformis polysaccharide degradation and its underlying mechanism involved in the Nrf-2/Keap-1 signaling pathway in HepG2 cells with oxidative stress induced by H₂O₂. The result of the scavenging ability of free radicals showed that GLP-HV (polysaccharide degraded by H₂O₂–vitamin C (Vc)) performed a better scavenging ability than GLP (G. lemaneiformis polysaccharide). Moreover, the scavenging ability of polysaccharide to these free radicals from strong to weak was as follows: superoxide radical, ferric ion, ABTS⁺, and DPPH radical, and their IC₅₀ values were 3.56 ± 0.0028, 4.97 ± 0.18, 9.62 ± 0.35, and 23.85 ± 1.78 mg/mL, respectively. Furthermore, GLP-HV obviously relieved oxidative stress in HepG2 cells, which strengthened the activity of T-AOC, CAT, GSH-PX, and SOD, and diminished the intensity of MDA, intracellular ROS, and calcium ion based on the Nrf-2/Keap-1 signaling pathway. The PCR result revealed that polysaccharide upregulated the expression of the genes Nrf-2, HO-1, NQO-1, and ZO-1 and downregulated Keap-1. The correlation between chemical properties and antioxidant mechanism of GLP-HV was evaluated via a heat map. The results illustrated that reducing sugar and active groups presented a positive correlation, and molecular weight and viscosity exhibited a negative relation with antioxidant activity.     

Astaxanthin Attenuates the Apoptosis of Retinal Ganglion Cells in db/db Mice by Inhibition of Oxidative Stress

Diabetic retinopathy is a common diabetic eye disease caused by changes in retinal ganglion cells (RGCs). It is an ocular manifestation of systemic disease, which affects up to 80% of all patients who have had diabetes for 10 years or more. The genetically diabetic db/db mouse, as a model of type-2 diabetes, shows diabetic retinopathy induced by apoptosis of RGCs. Astaxanthin is a carotenoid with powerful antioxidant properties that exists naturally in various plants, algae and seafood. Here, astaxanthin was shown to reduce the apoptosis of RGCs and improve the levels of oxidative stress markers, including superoxide anion, malondialdehyde (MDA, a marker of lipid peroxidation), 8-hydroxy-2-deoxyguanosine (8-OHdG, indicator of oxidative DNA damage) and MnSOD (manganese superoxide dismutase) activity in the retinal tissue of db/db mouse. In addition, astaxanthin attenuated hydrogen peroxide(H₂O₂)-induced apoptosis in the transformed rat retinal ganglion cell line RGC-5. Therefore, astaxanthin may be developed as an antioxidant drug to treat diabetic retinopathy.     

Chitosan Nanoparticles Attenuate Hydrogen Peroxide-Induced Stress Injury in Mouse Macrophage RAW264.7 Cells

This study was carried out to investigate the protective effects of chitosan nanoparticles (CNP) against hydrogen peroxide (H₂O₂)-induced oxidative damage in murine macrophages RAW264.7 cells. After 24 h pre-incubation with CNP (25–200 μg/mL) and chitosan (CS) (50–200 μg/mL, as controls), the viability loss in RAW264.7 cells induced by H₂O₂ (500 μM) for 12 h was markedly restored in a concentration-dependent manner as measured by MTT assay (P < 0.05) and decreased in cellular LDH release (P < 0.05). Moreover, CNP also exerted preventive effects on suppressing the production of lipid peroxidation such as malondialdehyde (MDA) (P < 0.05), restoring activities of endogenous antioxidant including superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) (P < 0.05), along with increasing total antioxidant capacity (T-AOC) (P < 0.05). In addition, pre-incubation of CNP with RAW264.7 cells for 24 h resulted in the increase of the gene expression level of endogenous antioxidant enzymes, such as MnSOD and GSH-Px (P < 0.05). At the same concentration, CNP significantly decreased LDH release and MDA (P < 0.05) as well as increased MnSOD, GSH-Px, and T-AOC activities (P < 0.05) as compared to CS. Taken together, our findings suggest that CNP can more effectively protect RAW264.7 cells against oxidative stress by H₂O₂ as compared to CS, which might be used as a potential natural compound-based antioxidant in the functional food and pharmaceutical industries.     

A Search for Hepatoprotective Activity of Fruit Extract of Mangifera indica L. Against Oxidative Stress Cytotoxicity

Mango (Mangifera indica L.) and their components are commonly used in folk medicine for many curative effects. The protective effects of different concentrations of aqueous extract of Mangifera indica L. fruit (Mango Extract) (20, 50 and 100 μg/ml) and also gallic acid (100 μM) as a pure compound in the extract were examined against oxidative stress toxicity induced by cumene hydroperoxide (CHP) in isolated rat hepatocytes. The extracts and gallic acid (100 μM) protected the hepatocyte against all oxidative stress markers including cell lysis, ROS generation, lipid peroxidation, glutathione depletion, mitochondrial membrane potential decrease, lysosomal membrane oxidative damage and cellular proteolysis. Mango Extracts (20, 50 and 100 μg/ml) were more effective than gallic acid (100 μM) in protecting hepatocytes against CHP induced lipid peroxidation. On the other hand gallic acid (100 μM) acted more effective than Mango Extracts (20, 50 and 100 μg/ml) at preventing lysosomal membrane damage. In addition H₂O₂ scavenging effect of all extracts were determined in hepatocytes and compared with gallic acid (100 μM). There were no significance differences (P<0.05) between all plant extracts and gallic acid (100 μM) in H₂O₂ scavenging activity. These results suggest a hepatoprotective role for Mango Extract against liver injury associated with oxidative stress.     

Ameliorating Effects of Fermented Rice Bran Extract on Oxidative Stress Induced by High Glucose and Hydrogen Peroxide in 3T3-L1 Adipocytes

In this study, we investigated whether fermented rice bran (FRB) can ameliorate the oxidative stress induced by high glucose and hydrogen peroxide (H₂O₂) in 3T3-L1 adipocytes by analyzing reactive oxygen species (ROS), oil red O staining, as well as the expression of mRNAs related to glucose homeostasis and adipogenesis. It was first confirmed that rice bran fermented by Issatchenkia orientalis MFST1 extract increased free phenolic content compared to non-fermented rice bran. The FRB extract strongly inhibited ROS generation and upregulated the expression of PPAR-γ and adiponectin. Moreover, FRB upregulated GLUT4 related to glucose transportation and insulin sensitivity. Taken together, FRB extract ameliorated oxidative stress-induced insulin resistance by neutralizing free radicals and upregulating adiponectin in adipocytes. Our results provide information toward understanding the beneficial effects of FRB on oxidative stress.     

White Tea (<Emphasis Type="Italic">Camellia sinensis</Emphasis> Kuntze) Exerts Neuroprotection against Hydrogen Peroxide-Induced Toxicity in PC12 Cells

Tea is a popular beverage whose consumption is associated with prevention of certain disorders. The objective of the study was to investigate the potential neuroprotective effect of white tea extract (WTE) on hydrogen peroxide induced toxicity in PC12 cells. Cells were treated with various doses of WTE (10–250 μg/ml) before exposition to 250 μM hydrogen peroxide and cell survival was determined through the MTT and LDH assays. Oxidative stress was quantified in the cells after treatments as intracellular reactive oxygen species (ROS) production and the antioxidant activity of the extract was assessed in a cell free system in terms of free radical scavenging capacity. Results showed that WTE has a significant protective effect in the PC12 cell line against hydrogen peroxide as cell survival was significantly superior in WTE-treated cells compared to hydrogen peroxide-treated cells. A reduction on intracellular oxidative stress as well as radical scavenging properties were produced by WTE. Results suggest that WTE protects PC12 cells against H₂O₂-induced toxicity, and that an antioxidant mechanism through ROS scavenging may be in part responsible for cells neuroprotection.     

Short-term evaluation of oxygen transfer from rice (Oryza sativa) to mixed planted drought-adapted upland crops under hydroponic culture

Mixed cropping is a cultivation method widely practiced in tropical regions. The newly developed close mixed planting technique mitigates the flood stress of drought-adapted upland cereal species by co-growing rice (Oryza sativa) plants under field flood conditions. We tested the hypothesis that O₂ was transferred from rice to upland crops using the model system of hydroponic culture. To confirm the hypothesis, the phenomena of O₂ absorption and release by plants were evaluated in a water culture condition without soil. Experiments were conducted in a climate chamber to estimate the amount of O₂ released from the roots of rice and pearl millet (Pennisetum glaucum) under both O₂-rich (20.0 ± .0% conc. in phase I) and O₂-free dark (.8 ± .0% conc. in phase II) conditions. The total O₂ change (between the two phases) in a single planting of rice and pearl millet was significantly higher than that of the mixed planting of rice and pearl millet, which indicated that O₂ was transferred from rice to pearl millet under a water culture condition. The result indicated that approximately 7 μM O₂ g fresh root weight^?1 h^?1 was transferred between the two plant species. O₂ transfer was confirmed between the two plant species in a mix cultured in water, implying its contribution to the phenomenon that improved the physiological status of drought-adapted upland crops under field flood conditions.     

The Cytoprotective Effect of Petalonia binghamiae Methanol Extract against Oxidative Stress in C2C12 Myoblasts: Mediation by Upregulation of Heme Oxygenase-1 and Nuclear Factor-Erythroid 2 Related Factor 2

This study was designed to examine the protective effects of the marine brown algae Petalonia binghamiae against oxidative stress-induced cellular damage and to elucidate the underlying mechanisms. P. binghamiae methanol extract (PBME) prevented hydrogen peroxide (H₂O₂)-induced growth inhibition and exhibited scavenging activity against intracellular reactive oxygen species (ROS) induced by H₂O₂ in mouse-derived C2C12 myoblasts. PBME also significantly attenuated H₂O₂-induced comet tail formation in a comet assay, histone γH2A.X phosphorylation, and annexin V-positive cells, suggesting that PBME prevented H₂O₂-induced cellular DNA damage and apoptotic cell death. Furthermore, PBME increased the levels of heme oxygenase-1 (HO-1), a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2 related factor 2 (Nrf2). However, zinc protoporphyrin IX, a HO-1 competitive inhibitor, significantly abolished the protective effects of PBME on H₂O₂-induced ROS generation, growth inhibition, and apoptosis. Collectively, these results demonstrate that PBME augments the antioxidant defense capacity through activation of the Nrf2/HO-1 pathway.     

Antioxidative Effect of Chlorella Pyrenoidosa Protein Hydrolysates and Their Application in Krill Oil-in-Water Emulsions

Chlorella pyrenoidosa is an excellent source of protein, and in this research, we assessed the antioxidant and emulsifying effects of Chlorella protein hydrolysate (CPH) using neutral proteases and alkaline proteases, as well as the properties of CPH-derived krill oil-in-water (O/W) emulsions. The CPHs exhibited the ability to scavenge several kinds of free radicals, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), O₂⁻, hydroxyl, and ABTS. Additionally, the CPHs (5 mg/mL) scavenged approximately 100% of the DPPH and ABTS. The CPHs showed similar emulsifying activities to Tween 20 and excellent foaming activities (max FS 74%), which helped to stabilize the krill oil-in-water emulsion. Less than 10 mg/mL CPHs was able to form fresh krill oil-in-water emulsions; moreover, the CPHs (5 mg/mL) in a krill O/W emulsion were homogenous, opaque, and stable for at least 30 days. Based on their inhibitory effects on the peroxide value (POV) and thiobarbituric acid reactive substances (TRABS), the CPHs were found to be able to inhibit lipid oxidation in both emulsifying systems and krill O/W emulsions. Thus, the CPHs could improve superoxide dismutase (SOD) activities by 5- or 10-fold and decrease the high reactive oxygen species (ROS) level caused by the addition of H₂O₂ in vitro. In conclusion, health-promoting CPHs could be applied in krill oil-in-water emulsions as both emulsifiers and antioxidants, which could help to improve the oxidative and physical stability of emulsions.     

外施海藻糖对高温胁迫下小麦幼苗膜脂过氧化的影响

在非生物胁迫条件下,海藻糖可以提高植物的抗逆性,减少逆境胁迫对植物组织的伤害,维持植物的相对正常生长。为了解其具体的作用机制,以小麦为材料,通过检测高温胁迫及室温恢复过程中丙二醛(MDA)含量、过氧化氢(H2O2)含量、超氧自由基(O·-2)含量、抗氧化物质[抗坏血酸(AsA)、还原型谷胱甘肽(GSH)]含量、抗氧化酶[超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)]活性及其基因转录的表达调控,以探究外源海藻糖对小麦幼苗抗氧化系统的影响。结果表明,在高温胁迫下外源海藻糖处理主要提高了AsA含量,增强了CAT和APX活性,同时上调了Mn-SOD、Cu/ZnSOD、CAT、POD和APX的相对表达量,从而降低了MDA及H2O2的产生。而随后的室温恢复过程基本与高温胁迫的结果一致,也提高了这些酶的基因转录水平以及AsA含量,主要差别是对抗氧化酶活性的影响,即室温恢复阶段外源海藻糖主要提高了POD和APX活性。综合来看,外源海藻糖在小麦幼苗的高温胁迫及室温恢复过程中,通过促进抗氧化酶基因的上调表达,提高抗氧化酶活性和抗氧化物质含量,以酶促和非酶促两种机制共同清除高温胁迫产生的活性氧,减少氧化胁迫的损伤,维持小麦幼苗的生长。     

Involvement of triadimefon induced early ABA-dependent H2O2 accumulation in soybean against water stress

The present study showed that pretreatment of triadimefon (TDM), a triazole compound, could improve tolerance of soybean seedlings to subsequent water stress. TDM pretreatment resulted in early and late rise in superoxide dismutase (SOD) and catalase (CAT) activities, and upregulation of ascorbate (AsA) content in non-stressed and water-stressed seedlings, leading to late increase in net photosynthetic rate (P_n), late decrease in hydrogen peroxide (H₂O₂) and electrolyte leakage in stressed ones. These TDM-induced changes were blocked by application of abscisic acid (ABA) biosynthesis inhibitor tungstate, which inhibited early rise of ABA and H₂O₂ contents in non-stressed and stressed seedlings. However, ABA pretreatment overcomed the effects of this inhibitor. Application of NADPH oxidase inhibitor diphenyleneiodonium (DPI), polyamine oxidase (PAO) inhibitor 2-hydroxyethylhydrazine (2-HEH) and H₂O₂ scavenger dimethylthiourea (DMTU) prevented early TDM-induced rise of H₂O₂ content. DPI, 2-HEH and DMTU also decreased SOD, CAT and AsA levels, but did not affect ABA content during early and late phases in both seedlings pretreated with TDM. In addition, these chemicals decreased P_n, and increased H₂O₂ content and electrolyte leakage during late phase in TDM-pretreated stressed seedlings. Overall, these results indicated that TDM pretreatment alleviated adverse effects of water stress on soybean seedlings, which was at least in part, due to increase of antioxidant capacity and decrease of oxidative damage induced by early ABA-dependent H₂O₂ generation.     

Comparative Analyses of Cu-Zn Superoxide Dismutase (SOD1) and Thioredoxin Reductase (TrxR) at the mRNA Level between Apis mellifera L. and Apis cerana F. (Hymenoptera: Apidae) Under Stress Conditions

This study compared stress-induced expression of Cu-Zn superoxide dismutase (SOD1) and thioredoxin reductase (TrxR) genes in the European honeybee Apis mellifera L. and Asian honeybee Apis cerana F. Expression of both SOD1 and TrxR rapidly increased up to 5 h after exposure to cold (4°C) or heat (37°C) treatment and then gradually decreased, with a stronger effect induced by cold stress in A. mellifera compared with A. cerana. Injection of stress-inducing substances (methyl viologen, [MV] and H₂O₂) also increased SOD1 and TrxR expression in both A. mellifera and A. cerana, and this effect was more pronounced with MV than H₂O₂. Additionally, we heterologously expressed the A. mellifera and A. cerana SOD1 and TrxR proteins in an Escherichia coli expression system, and detection by SDS-PAGE, confirmed by Western blotting using anti-His tag antibodies, revealed bands at 16 and 60 kDa, respectively. Our results show that the expression patterns of SOD1 and TrxR differ between A. mellifera and A. cerana under conditions of low or high temperature as well as oxidative stress.