Reduced Reactive Oxygen Species Accumulation Research Articles

Allelochemical p-hydroxybenzoic acid inhibits root growth via regulating ROS accumulation in cucumber (Cucumis sativus L.)

Allelopathy is prevalent in agricultural ecosystems and mediated by plant-derived secondary metabolites (allelochemicals). Allelochemicals are released by donor plants and affect the root growth and development of receptor plants. Allelopathy is responsible for the continuous cropping obstacles in cucumber (Cucumis sativus L.). p-Hydroxybenzoic acid (pHBA), an autotoxin from root exudates of cucumber, has been proposed to be an important allelopathic chemical. However, the molecular mechanism by which pHBA affect root growth and development in cucumber is unknown. Here, we found that pHBA treatment suppressed root growth of cucumber by reducing the meristem activity and cell length. This root growth defect is caused by reduced reactive oxygen species (ROS) accumulation in root tips. After pHBA treatment, the expression levels of several ROS-scavenging-related genes were increased, including peroxidase (POD), catalase (CAT) and metallothionein (MT). Moreover, exogenously application of salicylhydroxamate (SHAM), a peroxidase inhibitor, can partially restore the pHBA treatment induced root growth inhibition. Furthermore, we found that there is natural variation for the inhibitory effect of pHBA on root growth. We also showed that pHBA treatment could maintain higher level of ROS accumulated in the pHBA less sensitive cucumber than that in the pHBA-sensitive cucumber. These results suggest that pHBA inhibits root growth by reducing root tip ROS level in cucumber.

Reduced culture temperature attenuates oxidative stress and inflammatory response facilitating expansion and differentiation of adipose-derived stem cells

BackgroundAdipose-derived stem cell (ASC) expansion under atmospheric oxygen levels (21%) was previously shown to cause increased reactive oxygen species (ROS) accumulation and genetic instability compared to cells cultured under physiological oxygen levels (2–8%). However, since culture under physiological oxygen levels is costly and complicated, a simpler method to reduce ROS accumulation is desirable. The current study aimed to determine whether lower culture temperature can reduce ROS production in ASCs without impairing their culture expansion.MethodsProliferation, differentiation, ROS accumulation, and gene expression were compared between ASC cultures at 35 °C and 37 °C. ASCs isolated either from rat fat depots or from human lipoaspirates were examined in the study.ResultsRat visceral ASCs (vASCs) cultured at 35 °C demonstrated reduced ROS production and apoptosis and enhanced expansion and adipogenic differentiation compared to vASCs cultured at 37 °C. Similarly, the culture of human ASCs (hASCs) at 35 °C led to reduced ROS accumulation and apoptosis, with no effect on the proliferation rate, compared to hASCs cultured at 37 °C. Comparison of gene expression profiles of 35 °C versus 37 °C vASCs uncovered the development of a pro-inflammatory phenotype in 37 °C vASCs in correlation with culture temperature and ROS overproduction. This correlation was reaffirmed in both hASCs and subcutaneous rat ASCs.ConclusionsThis is the first evidence of the effect of culture temperature on ASC growth and differentiation properties. Reduced temperatures may result in superior ASC cultures with enhanced expansion capacities in vitro and effectiveness in vivo.

Puerarin Relieved Compression-Induced Apoptosis and Mitochondrial Dysfunction in Human Nucleus Pulposus Mesenchymal Stem Cells via the PI3K/Akt Pathway.

Puerarin (PUR), an 8-C-glucoside of daidzein extracted from Pueraria plants, is closely related to autophagy, reduced reactive oxygen species (ROS) production, and anti-inflammatory effects, but its effects on human nucleus pulposus mesenchymal stem cells (NPMSCs) have not yet been identified. In this study, NPMSCs were cultured in a compression apparatus to simulate the microenvironment of the intervertebral disc under controlled pressure (1.0 MPa), and we found that cell viability was decreased and apoptosis level was gradually increased as compression duration was prolonged. After PUR administration, apoptosis level evaluated by flow cytometry and caspase-3 activity was remitted, and protein levels of Bas as well as cleaved caspase-3 were decreased, while elevated Bcl-2 level was identified. Moreover, ATP production detection, ROS, and JC-1 fluorography as well as quantitative analysis suggested that PUR could attenuate intercellular ROS accumulation and mitochondrial dysfunction. Besides, the rat tail compression model was utilized, which indicated that PUR could restore impaired nucleus pulposus degeneration induced by compression. The PI3K/Akt pathway was identified to be deactivated after compression stimulation by western blot, and PUR could rescue the phosphorylation of Akt, thus reactivating the pathway. The effects of PUR, such as antiapoptosis, cell viability restoration, antioxidation, and mitochondrial maintenance, were all counteracted by application of the PI3K/Akt pathway inhibitor (LY294002). Summarily, PUR could alleviate compression-induced apoptosis and cell death of human NPMSCs in vitro as well as on the rat compression model and maintain intracellular homeostasis by stabilizing mitochondrial membrane potential and attenuating ROS accumulation through activating the PI3K/Akt pathway.

Hyperglycemia-induced endothelial dysfunction is alleviated by thioredoxin mimetic peptides through the restoration of VEGFR-2-induced responses and improved cell survival

BackgroundDiabetes mellitus is an important cardiovascular risk factor characterized by elevated plasma glucose levels. High glucose (HG) negatively influences endothelial cell (EC) function, which is characterized by the inability of ECs to respond to vascular endothelial growth factor (VEGF-A) stimulation. We aimed to identify potential strategies to improve EC function in diabetes. Methods and resultsHuman umbilical cord endothelial cells (HUVECs) were subjected to hyperglycemic milieu by exposing cells to HG together with glucose metabolite, methylglyoxal (MG) in vitro. Hyperglycemic cells showed reduced chemotactic responses towards VEGF-A as revealed by Boyden chamber migration assays, indicating the development of “VEGF resistance” phenotype. Furthermore, HG/MG-exposed cells were defective in their general migratory and proliferative responses and were in a pro-apoptotic state. Mechanistically, the exposure to HG/MG resulted in reactive oxygen species (ROS) accumulation which is secondary to the impairment of thioredoxin (Trx) activity in these cells. Pharmacological and genetic targeting of Trx recapitulated VEGF resistance. Functional supplementation of Trx using thioredoxin mimetic peptides (TMP) reversed the HG/MG-induced ROS generation, improved the migration, proliferation, survival and restored VEGF-A-induced chemotaxis and sprouting angiogenesis of hyperglycemic ECs. Importantly, TMP treatment reduced ROS accumulation and improved VEGF-A responses of placental arterial endothelial cells isolated from gestational diabetes mellitus patients. ConclusionsOur findings suggest a putative role for Trx in modulating EC function and its functional impairment in HG conditions contribute to EC dysfunction. Supplementation of TMP could be used as a novel strategy to improve endothelial cell function in diabetes.

Constitutive expression of the wheat TaSOD5 gene enhances salinity tolerance of Arabidopsis thaliana

Superoxide dismutase is a crucial reactive oxygen species (ROS) scavenger and converts the superoxide radical (O2-) to H2O2, so it is thought to enhance abiotic stress tolerance by reducing ROS accumulation and so avoiding oxidative damage. In this study, we isolated a salt- and oxidative stress-responsive Cu/Zn SOD gene TaSOD5 from wheat. The ectopic overexpression of TaSOD5 in Arabidopsis increased total and Cu/Zn SOD activities, and offered the plant tolerance to salt stress. Arabidopsis ectopically expressing TaSOD5 possessed a superior resistance to oxidative stress induced H2O2. The TaSOD5 ectopic overexpression elevated the activities of both ROS scavengers and O2- producer NADPH oxidase. These findings show that Cu/Zn SOD enhances salt tolerance via regulating the machinery of redox homeostasis rather than improving SOD activity alone.

Interactive Effects of Salicylic Acid and Nitric Oxide in Enhancing Rice Tolerance to Cadmium Stress.

Cadmium (Cd) is one of the prominent environmental hazards, affecting plant productivity and posing human health risks worldwide. Although salicylic acid (SA) and nitric oxide (NO) are known to have stress mitigating roles, little was explored on how they work together against Cd-toxicity in rice. This study evaluated the individual and combined effects of SA and sodium nitroprusside (SNP), a precursor of NO, on Cd-stress tolerance in rice. Results revealed that Cd at toxic concentrations caused rice biomass reduction, which was linked to enhanced accumulation of Cd in roots and leaves, reduced photosynthetic pigment contents, and decreased leaf water status. Cd also potentiated its phytotoxicity by triggering reactive oxygen species (ROS) generation and depleting several non-enzymatic and enzymatic components in rice leaves. In contrast, SA and/or SNP supplementation with Cd resulted in growth recovery, as evidenced by greater biomass content, improved leaf water content, and protection of photosynthetic pigments. These signaling molecules were particularly effective in restricting Cd uptake and accumulation, with the highest effect being observed in “SA + SNP + Cd” plants. SA and/or SNP alleviated Cd-induced oxidative damage by reducing ROS accumulation and malondialdehyde production through the maintenance of ascorbate and glutathione levels, and redox status, as well as the better activities of antioxidant enzymes superoxide dismutase, catalase, glutathione S-transferase, and monodehydroascorbate reductase. Combined effects of SA and SNP were observed to be more prominent in Cd-stress mitigation than the individual effects of SA followed by that of SNP, suggesting that SA and NO in combination more efficiently boosted physiological and biochemical responses to alleviate Cd-toxicity than either SA or NO alone. This finding signifies a cooperative action of SA and NO in mitigating Cd-induced adverse effects in rice, and perhaps in other crop plants.

WHIRLY1 Regulates HSP21.5A Expression to Promote Thermotolerance in Tomato.

Heat stress poses a major threat to plant productivity and crop yields. The induction of heat shock proteins (HSPs) by heat shock factors is a principal defense response of plants exposed to heat stress. In this study, we identified and analyzed the heat stress-induced Whirly1 (SlWHY1) gene in tomato (Solanum lycopersicum). We generated various SlWHY1-overexpressing (OE) and SlWHY1-RNA interference (RNAi) lines to investigate the role of WHIRLY1 in thermotolerance. Compared with the wild type (WT), the OE lines showed less wilting, as reflected by their increased membrane stability and soluble sugar content and reduced reactive oxygen species (ROS) accumulation under heat stress. By contrast, RNAi lines with inhibited SlWHY1 expression showed the opposite phenotype and corresponding physiological indices under heat stress. The heat-induced gene SlHSP21.5A, encoding an endoplasmic reticulum-localized HSP, was upregulated in the OE lines and downregulated in the RNAi lines compared with the WT. RNAi-mediated inhibition of SlHSP21.5A expression also resulted in reduced membrane stability and soluble sugar content and increased ROS accumulation under heat stress compared with the WT. SlWHY1 binds to the elicitor response element-like element in the promoter of SlHSP21.5A to activate its transcription. These findings suggest that SlWHY1 promotes thermotolerance in tomato by regulating SlHSP21.5A expression.

Biopriming with Piriformospora indica ameliorates cadmium stress in rice by lowering oxidative stress and cell death in root cells

Piriformospora indica is known for plant growth promotion and abiotic stress alleviation potential in several agricultural crops. However, a systemic analysis is warranted to explore potential application of this important fungus to augment heavy metal tolerance in rice. The present study explores potential of P. indica in ameliorating the effect of cadmium (Cd) stress in rice cultivars N22 and IR64. Seedlings inoculated with P. indica recorded significantly higher root-shoot length and biomass as compared to non-inoculated plants under control and Cd stress, respectively. Moreover, P. indica inoculated stressed roots accumulated more Cd as compared to non-inoculated stressed roots in both the varieties. Interestingly, cell death and reactive oxygen species (ROS) accumulation were significantly lower in the inoculated plant roots as compare with non-inoculated roots under Cd stress. The results emphasized significantly higher accumulation of Cd in fungal spores could reduce ROS accumulation in root cells resulting in lower cell death.

Boron mitigates citrus root injuries by regulating intracellular pH and reactive oxygen species to resist H+-toxicity

Boron (B)-deficiency and H+-toxicity are important limiting factors for plants growth in acid soils. High B supply may reduce H+-toxicity-induced inhibition of growth in citrus. Trifoliate orange rootstock seedlings were irrigated with nutrient solution containing either 0 μM or 10 μM H3BO3 at two pH levels (pH4 (H+-toxicity) and pH6 (normal)). The results showed that H+-toxicity without B severely hampered main root elongation. Simultaneously, oxidative damage caused by H+-toxicity led to severe damage to the apical structure of root such as root crown abscission. However, B application promoted the root length, root cell viability and reduced cell wall (CW) thickness of root tips under H+-toxicity. Additionally, B application reduced the H+-toxicity-induced reactive oxygen species (ROS) accumulation in roots as characterized by lower fluorescence intensity of H2O2 and O2- staining. Moreover, 31P-NMR (31P nuclear magnetic resonance) spectra revealed B application regulated the pH of vacuoles and cytoplasm in root tips by reducing phosphoenolpyruvate carboxykinase (PEPCase) activity while enhancing NADP malic enzyme (NADP-ME) activity during H+-toxicity. Collectively, our results demonstrate that B supply alleviates H+-toxicity and promotes root growth by reducing ROS accumulation, attenuating intracellular acidic microenvironment to ensure normal chemical reactions in root tip cells.

Excessive Accumulation of Ca2 + in Mitochondria of Y522S-RYR1 Knock-in Mice: A Link Between Leak From the Sarcoplasmic Reticulum and Altered Redox State.

Mice (Y522S or YS), carrying a mutation of the sarcoplasmic reticulum (SR) Ca2+ release channel of skeletal muscle fibers (ryanodine receptor type-1, RyR1) which causes Ca2+ leak, are a widely accepted and intensively studied model for human malignant hyperthermia (MH) susceptibility. Since the involvement of reactive oxygen species (ROS) and of mitochondria in MH crisis has been previously debated, here we sought to determine Ca2+ uptake in mitochondria and its possible link with ROS production in single fibers isolated from flexor digitorum brevis (FDB) of YS mice. We found that Ca2+ concentration in the mitochondrial matrix, as detected with the ratiometric FRET-based 4mtD3cpv probe, was higher in YS than in wild-type (WT) fibers at rest and after Ca2+ release from SR during repetitive electrical stimulation or caffeine administration. Also mitochondrial ROS production associated with contractile activity (detected with Mitosox probe) was much higher in YS fibers than in WT. Importantly, the inhibition of mitochondrial Ca2+ uptake achieved by silencing MCU reduced ROS accumulation in the matrix and Ca2+ release from SR. Finally, inhibition of mitochondrial ROS accumulation using Mitotempo reduced SR Ca2+ release in YS fibers exposed to caffeine. The present results support the view that mitochondria take up larger amounts of Ca2+ in YS than in WT fibers and that mitochondrial ROS production substantially contributes to the increased caffeine-sensitivity and to the enhanced Ca2+ release from SR in YS fibers.

Thioredoxin-1 confines T cell alloresponse and pathogenicity in graft-versus-host disease.

Oxidative stress is elevated in the recipients of allogeneic hematopoietic transplantation (allo-HCT) and likely contributes to the development of graft-versus-host disease (GVHD). GVHD is characterized by activation, expansion, cytokine production and migration of alloreactive donor T cells, and remains a major cause of morbidity and mortality after allo-HCT. Hence, strategies to limit oxidative stress in GVHD are highly desirable. Thioredoxin1 (Trx1) counteracts oxidative stress by scavenging reactive oxygen species (ROS) and regulating other enzymes that metabolize H2O2. The present study sought to elucidate the role of Trx1 in the pathophysiology of GVHD. Using murine and xenograft models of allogeneic bone marrow transplantation (allo-BMT) and genetic (human Trx1-transgenic, Trx1-Tg) as well as pharmacologic (human recombinant Trx1, RTrx1) strategies; we found that Trx1-Tg donor T cells or administration of the recipients with RTrx1 significantly reduced GVHD severity. Mechanistically, we observed RTrx1 reduced ROS accumulation and cytokine production of mouse and human T cells in response to alloantigen stimulation in vitro. In allo-BMT settings, we found that Trx1-Tg or RTrx1 decreased downstream signaling molecules including NFκB activation and T-bet expression, and reduced proliferation, IFN-γ production and ROS accumulation in donor T cells within GVHD target organs. More importantly, administration of RTrx1 did not impair the graft-versus-leukemia (GVL) effect. Taken together, the current work provides a strong rationale and demonstrates feasibility to target the ROS pathway, which can be readily translated into clinic.

Ebselen rescues oxidative-stress-suppressed osteogenic differentiation of bone-marrow-derived mesenchymal stem cells via an antioxidant effect and the PI3K/Akt pathway

BackgroundPatients with metabolic bone diseases often have high risk of titanium implant failure due to compromised bone regeneration ability. Clinical evidence indicates that the poor osteogenic ability is partly because of excessive oxidative stress. To date, specific treatments for these patients are urgently needed. Ebselen, a non-toxic organoselenium compound, is reported to be a potent antioxidant agent. In this study, we hypothesized that ebselen exerted protective effects on osteogenic differentiation of bone-marrow-derived mesenchymal stem cells (BMSCs) under oxidative stress. MethodsBMSCs were isolated from SD rats, and their morphology and multiple differentiation abilities were characterized. Proliferation rates of BMSCs treated with different concentrations of ebselen were analyzed. Then BMSCs were pretreated by hydrogen peroxide (H2O2), after which ebselen at different concentrations (0, 1, 5, 10 μM) was added, alkaline phosphatase (ALP) activity, mineralization and osteogenic-related protein levels were evaluated and an optimum concentration of ebselen was selected. Subsequently, intracellular reactive oxygen species (ROS) generation and the role of the PI3K/AKT pathway were also investigated. ResultsEbselen within a proper range could promote the proliferation of BMSCs. H2O2-induced oxidative stress suppressed osteogenic differentiation of BMSCs, which was verified by the decrease in ALP activity, calcium deposition, Runx2 and β-catenin expression. However, ebselen could alleviate osteogenic dysfunction of BMSCs. We also observed that ebselen reduced ROS accumulation in H2O2-pretreated BMSCs. Moreover, the pro-osteogenic effects afforded by ebselen were almost abolished by the Akt inhibitor. ConclusionWe concluded that ebselen could attenuate osteogenic dysfunction of BMSCs induced by H2O2 through an antioxidant effect and the activation of the PI3K/Akt pathway, suggesting that ebselen has a potential therapeutic effect for patients with metabolic bone diseases.

The Antioxidant Moiety of MitoQ Imparts Minimal Metabolic Effects in Adipose Tissue of High Fat Fed Mice.

Mitochondrial dysfunction is associated with a diverse array of diseases ranging from dystrophy and heart failure to obesity and hepatosteatosis. One of the major biochemical consequences of impaired mitochondrial function is an accumulation of mitochondrial superoxide, or reactive oxygen species (ROS). Excessive ROS can be detrimental to cellular health and is proposed to underpin many mitochondrial diseases. Accordingly, much research has been committed to understanding ways to therapeutically prevent and reduce ROS accumulation. In white adipose tissue (WAT), ROS is associated with obesity and its subsequent complications, and thus reducing mitochondrial ROS may represent a novel strategy for treating obesity related disorders. One therapeutic approach employed to reduce ROS abundance is the mitochondrial-targeted coenzyme Q (MitoQ), which enables mitochondrial specific delivery of a CoQ10 antioxidant via its triphenylphosphonium bromide (TPP+) cation. Indeed, MitoQ has been successfully shown to accumulate at the outer mitochondrial membrane and prevent ROS accumulation in several tissues in vivo; however, the specific effects of MitoQ on adipose tissue metabolism in vivo have not been studied. Here we demonstrate that mice fed high-fat diet with concomitant administration of MitoQ, exhibit minimal metabolic benefit in adipose tissue. We also demonstrate that both MitoQ and its control agent dTPP+ had significant and equivalent effects on whole-body metabolism, suggesting that the dTPP+ cation rather than the antioxidant moiety, was responsible for these changes. These findings have important implications for future studies using MitoQ and other TPP+ compounds.

Effects of exogenous spermidine and spermine on antioxidant metabolism associated with cold-induced leaf senescence in Zoysiagrass (Zoysia japonica Steud.)

Zoysiagrass (Zoysia japonica Steud.) leaves lose their green colour as they undergo cold acclimation (CA) in late fall in many regions, and delaying leaf senescence can extend their green period. The objective of this study was to investigate whether exogenous spermidine (Spd) and spermine (Spm) can delay leaf senescence by enhancing reactive oxygen species (ROS)-scavenging antioxidant enzyme activity. Mature zoysiagrass (cv. Zenith) plants were treated with Spm (1 mM) or Spd (1 mM) or water (control). The treated plants were grown under normal conditions or subjected to cold acclimation CA for 35 days. CA treatment increased leaf electrolyte leakage (EL) and malondialdehyde (MDA) level, decreased chlorophyll (Chl) content, and induced leaf senescence. Foliar application of Spd and Spm reduced the decline in Chl content and reduced EL and MDA levels. CA treatment increased the ROS level and reduced antioxidant enzyme activity. Application of Spm and Spd reduced ROS accumulation and enhanced the activities of superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase under CA conditions. The results of this study indicated that exogenous Spd and Spm at 1 mM can delay CA-induced leaf senescence through enhancement of ROS-scavenging antioxidant activity in zoysiagrass.

SoHSC70 positively regulates thermotolerance by alleviating cell membrane damage, reducing ROS accumulation, and improving activities of antioxidant enzymes

High temperature is a major environmental factor affecting plant growth. Heat shock proteins (Hsps) are molecular chaperones that play important roles in improving plant thermotolerance during heat stress. Spinach (Spinacia oleracea) is very sensitive to high temperature; however, the specific function of Hsps in spinach is unclear. In this study, cytosolic heat shock 70 protein (SoHSC70), which was induced by heat stress, was cloned from spinach. Overexpressing SoHSC70 in spinach calli and Arabidopsis enhanced their thermotolerance. In contrast, spinach seedlings with silenced SoHSC70 by virus-induced gene silencing (VIGS) showed more sensitivity to heat stress. Further analysis revealed that overexpressing SoHSC70 altered relative electrical conductivity (REC), malondialdehyde (MDA) content, photosynthetic rate, reactive oxygen species (ROS) accumulation and the activities of antioxidant enzymes, such as superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT) after the heat treatment. Taken together, our results suggest that overexpressing SoHSC70 positively affects heat tolerance by reducing membrane damage and ROS accumulation and improving activities of antioxidant enzymes.

Grafting improves tomato drought tolerance through enhancing photosynthetic capacity and reducing ROS accumulation.

Drought is the main meteorological threat to plants and limits plant growth, development, and adaptation to environmental changes. However, root-shoot communication plays a vital role in improving tomato plant drought tolerance, especially when cultivars are grafted onto drought-tolerant rootstock. In this study, the relationship between photosynthetic capacity and reactive oxygen species (ROS) in response to drought stress was studied in tomato grafted with different drought-resistant tomato seedlings. To determine the drought-relieving effect of drought-tolerant rootstocks, we measured the effects of grafting on plant growth, net photosynthetic rate (Pn), ROS accumulation, and antioxidant enzyme activities in tomato leaves and roots under drought stress. Plant growth and Pn were significantly inhibited by drought, but ROS accumulation and antioxidant enzyme activities were significantly increased. Treatment with drought-tolerant tomato seedlings significantly increased plant growth and increased Pn under water-deficit conditions compared with those grafted with drought-susceptible rootstock. In addition, the plants grafted with drought-tolerant seedlings had increased activities of partial antioxidant enzymes, leading to decreased ROS production. Our results indicate that tomato grafted with drought-tolerant seedlings alleviated the phytotoxicity and oxidative damage caused by drought by regulating antioxidant enzymes under drought stress.

Peroxisomal fatty acid β-oxidation negatively impacts plant survival under salt stress

ABSTRACTPeroxisomal β-oxidation is the sole pathway for metabolic breakdown of fatty acids to generate energy and carbon skeletons in plants, is essential for oilseed germination and plays an important role in growth, development and cellular homeostasis. Yet, this process also produces cytotoxic reactive oxygen species (ROS) as byproducts. We recently showed that disruption of fatty acid β-oxidation enhance plant survival under carbon starvation conditions. Here, we extend these findings by demonstrating that blocking fatty acid import into peroxisomes reduces ROS accumulation and increases plant tolerance to salt stress, whereas increasing fatty acid flux into the β-oxidation pathway has opposite effects. Together, these results support the view that peroxisomal β-oxidation of fatty acids enhances stress-induced ROS production, thereby negatively impacting plant survival under adverse environmental conditions.

Linking monoterpenes and abiotic stress resistance in grapevines

Rising temperatures and ozone levels are among the most striking stressful phenomena of global climate changes, and they threaten plants that are unable to react rapidly and efficiently. Generic responses of plants to stresses include the production of excess reactive oxygen species (ROS). Excessive ROS accumulation can lead to extensive oxidation of important components such as nucleic acids, proteins and lipids which can further exacerbate ROS accumulation leading to programmed cell death. Although most studies on plant antioxidants have focused on non-volatile compounds, volatiles belonging to the isoprenoid family have been implicated in the protection against abiotic stresses, in particular thermal and oxidative stress whose frequency and extent is being exacerbated by ongoing global change and anthropogenic pollution. Historically, research has focused on isoprene, demonstrating that isoprene-emitting plants are more tolerant to ozone exposure and heat stress, reducing ROS accumulation. Yet, evidence is being compiled that shows other volatile isoprenoids may be involved in plant responses against abiotic stresses. Grapevines are not isoprene emitters but some varieties produce other volatile isoprenoids such as monoterpenes. We investigated photosynthesis and emission of volatile organic compounds upon heat stress in two Vitis vinifera cv. ‘Chardonnay’ clones differing only for a mutation in the DXS gene (2-C-methyl-D-erythritol 4-phosphate (MEP) pathway), regulating volatile isoprenoid biosynthesis. We showed that the mutation led to a strong increase in monoterpene emission upon heat stress. At the same time, maximum photochemical quantum yield (Fv/Fm ratio) of PSII was affected by the stress in the non-emitting clone while the monoterpene emitter showed a significant resilience, thus indicating a possible antioxidant role of monoterpenes in grapevine. Future mechanistic studies should focus on unveiling the actual mechanism responsible for such findings.

Resveratrol Alleviates Rheumatoid Arthritis via Reducing ROS and Inflammation, Inhibiting MAPK Signaling Pathways, and Suppressing Angiogenesis

Rheumatoid arthritis (RA) is a systemic autoimmune disease primarily affecting joints and is featured by chronic synovial inflammation and angiogenesis. We employed a bovine type-II collagen (BIIC)-induced Sprague-Dawley rat arthritis model and an in vitro RA model based on interleukin (IL)-1β-stimulated rat synovial cells (RSC-364) to explore the preventive effect of resveratrol on RA and the underlying mechanisms. We found that resveratrol ameliorated BIIC-elicited synovitis and RA-related pathological hallmarks such as inflammatory cell infiltration and angiogenesis in the synovial tissue. Also, BIIC-stimulated rats displayed increased serum levels of proinflammatory cytokines and reactive oxygen species (ROS), as manifested by elevated serum malonaldehyde contents combined with reduced superoxide dismutase activity. It is noteworthy that resveratrol abolished BIIC-induced ROS and inflammation, confirming the antioxidative and anti-inflammatory actions of resveratrol in the context of RA. Furthermore, immunoblotting indicated that resveratrol downregulated the increase in the levels of hypoxia-inducible factor-1α (HIF-1α) and that of the activated phosphorylation of p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase in IL-1β-stimulated RSC-364 cells. Moreover, we observed that resveratrol-treated RSC-364 cells displayed both G0/G1 cell-cycle arrest and enhanced levels of apoptosis. Altogether, the present evidence established the preventive role of resveratrol in RA progression. Mechanistically, resveratrol inhibits MAPK signaling pathways, likely by reducing ROS accumulation, to suppress the inflammatory response and cell proliferation and to provoke cell apoptosis in the synovial tissue, along with mitigation of HIF-1α-mediated angiogenesis. Thus resveratrol appears to hold great potential for clinical translation as a novel RA therapeutic.

RBOH1-dependent apoplastic H2O2 mediates epigallocatechin-3-gallate-induced abiotic stress tolerance in Solanum lycopersicum L.

Responses of plants to abiotic stimuli are mediated by a plethora of bioactive compounds. Flavonoids are a large group of plant secondary metabolites, playing diverse roles in plant growth, development and responses to stress. Despite the recent report on epigallocatechin-3-gallate (EGCG, a bioactive flavonoids)-induced enhanced tolerance to salt stress, the underlying molecular mechanisms of stress tolerance remain unclear. Moreover, whether such stress protective role of EGCG is common for other environmental stressors also remains elusive. In the current study, we showed that EGCG functions as a broad spectrum stress protectant that alleviates a variety of abiotic stresses such as cold, heat, salinity and drought in tomato plants. While those abiotic stresses inhibited leaf photosynthetic activity by enhancing reactive oxygen species (ROS) production and membrane damage, exogenous EGCG protected photosynthetic apparatus by reducing ROS accumulation. Furthermore, EGCG-induced attenuation of ROS accumulation was largely attributed to the enhanced activity of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX). Analysis of sub-cellular localization of ROS revealed that EGCG preferentially induced apoplastic H2O2 production by activation of RESPIRATORY BURST OXIDASE HOMOLOG1 (RBOH1)-encoded NADPH oxidase. Further investigation using virus-induced gene silencing showed that suppression of RBOH1 expression aggravated abiotic stress-induced ROS accumulation and compromised EGCG-induced ROS scavenging, implying that RBOH1-dependent H2O2 production is critical for the EGCG-induced tolerance to abiotic stress. This study advances our current understanding of stress protective role of flavonoids and unveils a critical mechanism of EGCG-induced abiotic stress tolerance in tomato plants.