High Levels Of Hydrogen Peroxide Research Articles

Selective targeting of cancer cells using a hydrogen peroxide-activated Hsp90 inhibitor

Heat shock protein 90 (Hsp90) plays an important role in cancer cell proliferation, survival, and migration by regulating the maturation and stabilization of numerous oncoproteins. Despite significant efforts in developing Hsp90 inhibitors, none of these have been approved for clinical use, mostly due to toxicity, such as liver, cardiac, and retinal toxicity. To avoid undesirable toxicity, we herein report a hydrogen peroxide-activated Hsp90 inhibitor, Boro-BZide (3), which is capable of selectively targeting cancer cells over normal cells. Boro-BZide (3) can be activated by high levels of hydrogen peroxide, releasing its parent active Hsp90 inhibitor. The mechanism of action was determined by a series of experiments including fluorescence polarization assay, cell viability assay, western blotting, high-pressure liquid chromatography (HPLC), and fluorescence-activated cell sorting (FACS) analysis. These efforts ultimately led to the identification of a novel hydrogen peroxide-activated Hsp90 prodrug with improved therapeutic index, which was less prone to furnish unwanted adverse effects. This hydrogen peroxide-responsive prodrug strategy will be beneficial for overcoming the toxicity hurdles of Hsp90 inhibitors for clinical application.

Glyoxylate reductase/hydroxypyruvate reductase regulates the free d-aspartate level in mammalian cells.

Multiple d-amino acids are present in mammalian cells, and these compounds have distinctive physiological functions. Among the free d-amino acids identified in mammals, d-aspartate plays critical roles in the neuroendocrine and endocrine systems, as well as in the central nervous system. Mammalian cells have the molecular apparatus necessary to take up, degrade, synthesize, and release d-aspartate. In particular, d-aspartate is degraded by d-aspartate oxidase (DDO), a peroxisome-localized enzyme that catalyzes the oxidative deamination of d-aspartate to generate oxaloacetate, hydrogen peroxide, and ammonia. However, little is known about the molecular mechanisms underlying d-aspartate homeostasis in cells. In this study, we established a cell line that overexpresses cytoplasm-localized DDO; this cell line cannot survive in the presence of high concentrations of d-aspartate, presumably because high levels of toxic hydrogen peroxide are produced by metabolism of abundant d-aspartate by DDO in the cytoplasm, where hydrogen peroxide cannot be removed due to the absence of catalase. Next, we transfected these cells with a complementary DNA library derived from the human brain and screened for clones that affected d-aspartate metabolism and improved cell survival, even when the cells were challenged with high concentrations of d-aspartate. The screen identified a clone of glyoxylate reductase/hydroxypyruvate reductase (GRHPR). Moreover, the GRHPR metabolites glyoxylate and hydroxypyruvate inhibited the enzymatic activity of DDO. Furthermore, we evaluated the effects of GRHPR and peroxisome-localized DDO on d- and l-aspartate levels in cultured mammalian cells. Our findings show that GRHPR contributes to the homeostasis of these amino acids in mammalian cells.

MeRAV5 promotes drought stress resistance in cassava by modulating hydrogen peroxide and lignin accumulation.

Cassava, an important food and energy crop, is relatively more resistant to drought stress than other crops. However, the molecular mechanism underlying this resistance remains elusive. Herein, we report that silencing a drought stress-responsive transcription factor MeRAV5 significantly reduced drought stress resistance, with higher levels of hydrogen peroxide (H2 O2 ) and less lignin during drought stress. Yeast two-hybrid, pull down and bimolecular fluorescence complementation (BiFC) showed that MeRAV5 physically interacted with peroxidase (MePOD) and lignin-related cinnamyl alcohol dehydrogenase 15 (MeCAD15) invitro and invivo. MeRAV5 promoted the activities of both MePOD and MeCAD15 to affect H2 O2 and endogenous lignin accumulation respectively, which are important in drought stress resistance in cassava. When either MeCAD15 or MeRAV5 was silenced, or both were co-silenced, cassava showed lower lignin content and drought-sensitive phenotype, whereas exogenous lignin alkali treatment increased drought stress resistance and alleviated the drought-sensitive phenotype of these silenced cassava plants. This study documents that the modulation of H2 O2 and lignin by MeRAV5 is essential for drought stress resistance in cassava.

The Use of Topical Estrogen Supplementation to Combat Arsenic‐Induced Redox Stress in Ovariectomized Murine Models

Background Chronic environmental arsenic exposure is a health hazard that can lead to cancer, cardiovascular disease and diabetes. Arsenic exposure stimulates the production of reactive oxygen species (ROS) increasing systemic oxidative stress. Estrogen contains antioxidant properties that assist in the upregulation of various antioxidant enzymes, including glutathione. The present study investigated the effects of estrogen as a therapeutic agent to combat oxidative stress caused by arsenic exposure in ovariectomized murine models. Methods Ovariectomized mice (10 weeks old) (n=24) were randomized to four treatment groups (CON= control, CE=control + estrogen, ARS=arsenic, AE= arsenic + estrogen). Arsenic treated groups (ARS & AE) were exposed to controlled arsenic levels (10µM) via drinking water for 3-wks. Following the arsenic exposure, estrogen supplementation (0.75 µg/g of 0.06% Estradiol Benzoate) was applied topically to respective treatment groups (CE & AE). One week following the first estrogen dosage (3 total doses), mice were euthanized, and skeletal muscle and cardiac tissue were collected. Oxidized and reduced glutathione and catalase (CAT) activity were measured in skeletal muscle. Superoxide dismutase (SOD) and thioredoxin reductase (TrxR) were measured in cardiac tissue. A One-way ANOVA was performed with post-hoc comparisons using Least Significant Differences (LSD) for pairwise comparisons. Results For reduced glutathione levels, CE was significantly higher than CON (p = 0.004) and ARS (p = 0.001). CE was also significantly higher than all other groups for GSSG (p < 0.05) for all pairwise comparisons). Catalase activity was significantly higher in AE compared to CON (p = 0.032), and CE (p = 0.001), and ARS (p = 0.009). Superoxide dismutase (SOD) and thioredoxin reductase (TrxR) levels did not differ across treatment groups. Conclusion Estrogen treatment increased total glutathione content in the CE group indicated by the high levels of GSH as well as GSSG. Estrogen treatment combined with the arsenic exposure also increased catalase activity levels pointing to higher levels of hydrogen peroxide in the AE group, although this was not measured directly. Taken together, these data suggest estrogen may increase antioxidant capacity by elevating total glutathione and catalase content within skeletal muscle. These data indicate that estrogen alone and estrogen in the presence arsenic may play a role in modulating antioxidant activity in arsenic-induced oxidative damage and may be a potential treatment option for individuals suffering from environmental arsenic exposure.

Differential oxidative stress responses and tobacco-specific nitrosamine accumulation in two burley varieties

Tobacco-specific nitrosamines (TSNAs) are carcinogens that accumulate in tobacco leaves during curing, storage, and processing, and their amounts in processed tobacco vary dependent on several intrinsic and extrinsic factors. Here, we assessed the hypothesis that there is a link between reactive oxygen species levels in leaves and TSNA formation during curing. First, we show that burley varieties KT 204LC and NCBH 129LC accumulate TSNAs to different levels but not as a result of a variety-specific abundance of TSNA precursors. Next, we measured the levels of reactive oxygen species, and we show that the variety that accumulates more TSNAs, NCBH 129LC, had significantly higher levels of hydrogen peroxide than KT 204LC. The NCBH 129LC also has more oxidatively damaged and glutathionylated proteins. Finally, we analyzed the antioxidant levels in KT 204LC and NCBH 129LC and their tolerance to oxidative stress. NCBH 129LC contained more of the essential antioxidant glutathione and was more tolerant to the oxidative stress-generating compound paraquat. Collectively, our data suggest that there is indeed a link between foliar oxidative stress parameters and the extent to which TSNAs accumulate in cured tobacco leaves.

Chloroplast thylakoid ascorbate peroxidase PtotAPX plays a key role in chloroplast development by decreasing hydrogen peroxide in Populus tomentosa.

Chloroplast development is a complex process that is critical to the growth and development of plants. However, the detailed mechanism of chloroplast development in woody plants remains unclear. In this study, we showed that chloroplasts with elaborate thylakoids could develop from proplastids in the cells of calli derived from leaf tissues of Populus tomentosa upon exposure to light. Chloroplast development was confirmed at the molecular and cellular levels. Transcriptome analysis revealed that genes related to photoreceptors and photosynthesis were significantly up-regulated during chloroplast development in a time-dependent manner. In light-induced chloroplast development, a key process was the removal of hydrogen peroxide, in which thylakoid-localized PtotAPX played a major role; light-induced chloroplast development was enhanced in PtotAPX-overexpressing transgenic P. tomentosa callus with lower levels of hydrogen peroxide, but was suppressed in PtotAPX antisense transgenic callus with higher levels of hydrogen peroxide. Moreover, the suppression of light-induced chloroplast development in PtotAPX antisense transgenic callus was relieved by the exogenous reactive oxygen species scavenging agent N,N'-dimethylthiourea (DMTU). Based on these results, we propose that PtotAPX-mediated removal of reactive oxygen species plays a key role in chloroplast development from proplastids upon exposure to light in P. tomentosa.

Zinc Oxide Nanoparticles and Zinc Sulfate Impact Physiological Parameters and Boosts Lipid Peroxidation in Soil Grown Coriander Plants (Coriandrum sativum).

The objective of this study was to determine the oxidative stress and the physiological and antioxidant responses of coriander plants (Coriandrum sativum) grown for 58 days in soil with zinc oxide nanoparticles (ZnO NPs) and zinc sulfate (ZnSO4) at concentrations of 0, 100, 200, 300, and 400 mg of Zn/kg of soil. The results revealed that all Zn compounds increased the total chlorophyll content (CHLt) by at least 45%, compared to the control group; however, with 400 mg/kg of ZnSO4, chlorophyll accumulation decreased by 34.6%. Zn determination by induction-plasma-coupled atomic emission spectrometry (ICP–AES) showed that Zn absorption in roots and shoots occurred in plants exposed to ZnSO4 at all concentrations, which resulted in high levels of hydrogen peroxide (H2O2) and malondialdehyde (MDA). Only at 400 mg/kg of ZnSO4, a 78.6% decrease in the MDA levels was observed. According to the results, the ZnSO4 treatments were more effective than the ZnO NPs to increase the antioxidant activity of catalase (CAT), ascorbate peroxidase (APX), and peroxidases (POD). The results corroborate that phytotoxicity was higher in plants subjected to ZnSO4 compared to treatments with ZnO NPs, which suggests that the toxicity was due to Zn accumulation in the tissues by absorbing dissolved Zn++ ions.

Ascorbate-glutathione pathways mediated by cytokinin regulate H2O2 levels in light-controlled rose bud burst.

Rosebush (Rosa “Radrazz”) plants are an excellent model to study light control of bud outgrowth since bud outgrowth only arises in the presence of light and never occurs in darkness. Recently, we demonstrated high levels of hydrogen peroxide (H2O2) present in the quiescent axillary buds strongly repress the outgrowth process. In light, the outgrowing process occurred after H2O2 scavenging through the promotion of Ascorbic acid–Glutathione (AsA–GSH)-dependent pathways and the continuous decrease in H2O2 production. Here we showed Respiratory Burst Oxidase Homologs expression decreased in buds during the outgrowth process in light. In continuous darkness, the same decrease was observed although H2O2 remained at high levels in axillary buds, as a consequence of the strong inhibition of AsA–GSH cycle and GSH synthesis preventing the outgrowth process. Cytokinin (CK) application can evoke bud outgrowth in light as well as in continuous darkness. Furthermore, CKs are the initial targets of light in the photocontrol process. We showed CK application to cultured buds in darkness decreases bud H2O2 to a level that is similar to that observed in light. Furthermore, this treatment restores GSH levels and engages bud burst. We treated plants with buthionine sulfoximine, an inhibitor of GSH synthesis, to solve the sequence of events involving H2O2/GSH metabolisms in the photocontrol process. This treatment prevented bud burst, even in the presence of CK, suggesting the sequence of actions starts with the positive CK effect on GSH that in turn stimulates H2O2 scavenging, resulting in initiation of bud outgrowth.

Nitric Oxide (NO) Scaffolds the Peroxisomal Protein-Protein Interaction Network in Higher Plants.

The peroxisome is a single-membrane subcellular compartment present in almost all eukaryotic cells from simple protists and fungi to complex organisms such as higher plants and animals. Historically, the name of the peroxisome came from a subcellular structure that contained high levels of hydrogen peroxide (H2O2) and the antioxidant enzyme catalase, which indicated that this organelle had basically an oxidative metabolism. During the last 20 years, it has been shown that plant peroxisomes also contain nitric oxide (NO), a radical molecule than leads to a family of derived molecules designated as reactive nitrogen species (RNS). These reactive species can mediate post-translational modifications (PTMs) of proteins, such as S-nitrosation and tyrosine nitration, thus affecting their function. This review aims to provide a comprehensive overview of how NO could affect peroxisomal metabolism and its internal protein-protein interactions (PPIs). Remarkably, many of the identified NO-target proteins in plant peroxisomes are involved in the metabolism of reactive oxygen species (ROS), either in its generation or its scavenging. Therefore, it is proposed that NO is a molecule with signaling properties with the capacity to modulate the peroxisomal protein-protein network and consequently the peroxisomal functions, especially under adverse environmental conditions.

Streptococcus pneumoniae and Its Virulence Factors H2O2 and Pneumolysin Are Potent Mediators of the Acute Chest Syndrome in Sickle Cell Disease.

Sickle cell disease (SCD) is one of the most common autosomal recessive disorders in the world. Due to functional asplenia, a dysfunctional antibody response, antibiotic drug resistance and poor response to immunization, SCD patients have impaired immunity. A leading cause of hospitalization and death in SCD patients is the acute chest syndrome (ACS). This complication is especially manifested upon infection of SCD patients with Streptococcus pneumoniae (Spn)—a facultative anaerobic Gram-positive bacterium that causes lower respiratory tract infections. Spn has developed increased rates of antibiotics resistance and is particularly virulent in SCD patients. The primary defense against Spn is the generation of reactive oxygen species (ROS) during the oxidative burst of neutrophils and macrophages. Paradoxically, Spn itself produces high levels of the ROS hydrogen peroxide (H2O2) as a virulence strategy. Apart from H2O2, Spn also secretes another virulence factor, i.e., the pore-forming exotoxin pneumolysin (PLY), a potent mediator of lung injury in patients with pneumonia in general and particularly in those with SCD. PLY is released early on in infection either by autolysis or bacterial lysis following the treatment with antibiotics and has a broad range of biological activities. This review will discuss recent findings on the role of pneumococci in ACS pathogenesis and on strategies to counteract the devastating effects of its virulence factors on the lungs in SCD patients.

Interference of CsGPA1, the α-submit of G protein, reduces drought tolerance in cucumber seedlings

The G protein α-subunit, GPA1, is an integral component of several signaling pathways in plants, including response to abiotic stress. However, the molecular mechanism behind these processes remains largely unknown in the cucumber plant (Cucumis sativus L.). In order to further understand the role of CsGPA1 in cucumber under drought stress, changes in plant growth, physiological parameters, and gene expression of CsAQPs were all measured under water stress induced by polyethylene glycol (PEG) using wild type (WT) and CsGPA1-interference (RNAi) cucumber seedlings. Our results demonstrated that the RNAi plants had lower drought tolerance, displaying seriously withered leaves, lower relative growth rate, lower root-shoot ratio, and lower root activity under drought stress compared to WT plants. Physiological studies indicated that the suppression of CsGPA1 weakened drought stress tolerance due to higher water loss rate in the leaves, higher levels of hydrogen peroxide (H2O2), increased malondialdehyde (MDA) content, lower free proline content, lower soluble sugar content, lower soluble protein content, and decreased antioxidant enzyme activities. qRT-PCR analysis demonstrated that the interference of CsGPA1 up-regulated the expression of most AQP genes (except for CsPIP2;3 in leaves) and down-regulated the expression of CsPIP1;2, CsPIP1;4, CsPIP2;1, and CsPIP2;4 in roots under drought stress when compared to WT plants. Our results demonstrated that CsGPA1 could function as a positive regulator in drought stress response by decreasing the accumulation of reactive oxygen species (ROS), improving permeable potentials, and reducing water loss in cucumber plants.

Gly Betaine Surpasses Melatonin to Improve Salt Tolerance in Dalbergia odorifera.

Salinity is one of the most serious factors limiting plant growth which can provoke significant losses in agricultural crop production, particularly in arid and semi-arid areas. This study aimed to investigate whether melatonin (MT; 0.05 and 0.1 mM), which has pleiotropic roles, has a better effect than glycine betaine (GB; 10 and 50 mM) on providing salt tolerance in a woody plant Dalbergia odorifera T. Chen. Also, the alternative oxidase activity (AOX) in plant subjected to MT or GB under salinity (150 and 250 mM) was evaluated given that the effect of exogenous MT or GB on AOX has not been reported yet. The results showed that the exogenous application of GB on the seedlings of D. odorifera increased the plant growth parameters, relative water content, total of chlorophyll content, and carotenoid content compared with well-watered and MT treatments. Under severe salinity, the seedlings subjected to GB showed, a significant enhancement in water use efficiency, transpiration, and net photosynthetic rate regardless to MT-treated seedlings. The levels of proline and soluble sugar in the seedlings treated with MT or GB decreased significantly under mild and severe salinity correlated with those in salt-stressed seedlings. Furthermore, GB-treated plants exhibited a significant inhibition of malondialdehyde content compared with MT-treated plants. The concentration of thiols and phenolic compounds were significantly enhanced in the leaves of seedlings treated with MT compared with those treated with GB. Under salt stress condition, GB scavenged significantly higher levels of hydrogen peroxide than MT; while under severe salinity, plants subjected to MT showed better scavenging ability for hydroxyl radicals compared with GB-treated seedlings. The results demonstrated also an enhancement of the levels of superoxide dismutase (SOD), guaiacol peroxidase, and AOX activities in seedlings treated with GB or MT compared with salt-stressed plants. The catalase activity (CAT) was increased by 0.05 mM MT and 0.1 mM GB under mild salinity. Meanwhile, the AOX activity under severe salinity was enhanced only by GB 50 mM. The findings of this study suggested that GB-treated seedlings possessed a better salt tolerance in comparison with MT-treated seedlings.

Thioredoxin pathway in Anabaena sp. PCC 7120: activity of NADPH-thioredoxin reductase C.

To understand the physiological role of NADPH-thioredoxin reductase C (NTRC) in cyanobacteria, we investigated an NTRC-deficient mutant strain of Anabaena sp., PCC 7120, cultivated under different regimes of nitrogen supplementation and light exposure. The deletion of ntrC did not induce a change in the cell structure and metabolic pathways. However, time-dependent changes in the abundance of specific proteins and metabolites were observed. A decrease in chlorophyll a was correlated with a decrease in chlorophyll a biosynthesis enzymes and photosystem I subunits. The deletion of ntrC led to a deregulation of nitrogen metabolism, including the NtcA accumulation and heterocyst-specific proteins while nitrate ions were available in the culture medium. Interestingly, this deletion resulted in a redox imbalance, indicated by higher peroxide levels, higher catalase activity and the induction of chaperones such as MsrA. Surprisingly, the antioxidant protein 2-CysPrx was downregulated. The deficiency in ntrC also resulted in the accumulation of metabolites such as 6-phosphogluconate, ADP and ATP. Higher levels of NADP+ and NADPH partly correlated with higher G6PDH activity. Rather than impacting protein expression levels, NTRC appears to be involved in the direct regulation of enzymes, especially during the dark-to-light transition period.

An Ultrasmall SnFe2O4 Nanozyme with Endogenous Oxygen Generation and Glutathione Depletion for Synergistic Cancer Therapy

AbstractThe tumor microenvironment (TME) with the characteristics of severe hypoxia, overexpressed glutathione (GSH), and high levels of hydrogen peroxide (H2O2) dramatically limits the antitumor efficiency by monotherapy. Herein, a novel TME‐modulated nanozyme employing tin ferrite (SnFe2O4, abbreviated as SFO) is presented for simultaneous photothermal therapy (PTT), photodynamic therapy (PDT), and chemodynamic therapy (CDT). The as‐fabricated SFO nanozyme demonstrates both catalase‐like and GSH peroxidase‐like activities. In the TME, the activation of H2O2 leads to the generation of hydroxyl radicals (•OH) in situ for CDT and the consumption of GSH to relieve antioxidant capability of the tumors. Meanwhile, the nanozyme can catalyze H2O2 to generate oxygen to meliorate the tumor hypoxia, which is beneficial to achieve better PDT. Furthermore, the SFO nanozyme irradiated with 808 nm laser displays a prominent phototherapeutic effect on account of the enhanced photothermal conversion efficiency (η = 42.3%) and highly toxic free radical production performance. This “all in one” nanozyme integrated with multiple treatment modalities, computed tomography, and magnetic resonance imaging properties, and persistent modulation of TME exhibits excellent tumor theranostic performance. This strategy may provide a new dimension for the design of other TME‐based anticancer strategies.

Chemiluminescent Measurement of Hydrogen Peroxide in the Exhaled Breath Condensate of Healthy and Asthmatic Adults.

Reactive oxygen species are centrally involved in the pathophysiology of airway diseases such as asthma and chronic obstructive pulmonary disease. This study reports the development of a chemiluminescence assay and a device for measuring hydrogen peroxide in the exhaled breath condensate of asthma patients and healthy participants. A stand-alone photon detection device was constructed for use with an optimized chemiluminescence assay. Calibrations using a catalase control to scavenge residual hydrogen peroxide in calibrant solutions provided analytically sensitive and specific measurements. We evaluated exhaled breath condensate hydrogen peroxide in 60 patients (ages 20-83; 30 healthy patients and 30 asthma patients) recruited from the John Peter Smith Hospital Network. The exhaled breath condensate hydrogen peroxide concentrations trended toward higher values in asthma patients compared to healthy participants (mean 142.5 vs 115.5 nM; p = 0.32). Asthma patients who had not used an albuterol rescue inhaler in the past week were compared to those who had and showed a trend toward higher hydrogen peroxide levels (mean 172.8 vs 115.9 nM; p = 0.25), and these patients also trended toward higher hydrogen peroxide than healthy participants (mean 172.8 vs 115.5 nM; p = 0.14). This pilot study demonstrates the ability of the newly developed assay and device to measure exhaled breath condensate hydrogen peroxide in asthma patients and healthy participants. The trends observed in this study are in agreement with previous literature and warrant further investigation of using this system to measure exhaled breath condensate hydrogen peroxide for monitoring oxidative stress in asthma.

CHARACTERIZATION OF PHYSICOCHEMICAL PROPERTIES AND BIOACTIVE COMPOUNDS OF OIL FROM NASCENT SEEDS OF AZADIRACHTA INDICA

Depending on composition, fatty acids of plant sources are explicitly useful in food, chemical and pharmaceutical industries. The present study characterizes Soxhlet extracted neem seed oil by determination of physicochemical properties and bioactive compounds. Extraction involved the use of solvent semi-continuous extraction method for 6 hours with N-hexane. Percentage Oil yield was at 39. Saponification value (50.490mgKOH/g), Iodine value (50.28g/100g), Acid value (5.610mgKOH/g), Free fatty acid (2.805w/w), Peroxide value (62.75meq/kg) and Specific gravity (0.948) were determined. Density at 250C, Flash point, boiling point, melting point, colour and Odour provided supportive information to the major physicochemical properties analyzed. Gas Chromatography - Mass Spectrometry (GC-MS) of the Soxhlet extracted Neem Seed oil showed different peak values of industrially essential compounds such as 7-Pentadecyne, Glycidyl palmitate, Oleic Acid, 9,12-Octadecadienoic acid and methyl ester (Omega-6 acid). Saponification value was found to be high, suggesting the presence of FFA with low molecular weight, which brands the oil as essential in cosmetic and food industries. Low Iodine value and low total acidity seems to consolidate the usefulness of neem seed oil in cosmetic industries. Conversely, the high peroxide level makes it unsuitable for cooking. It may therefore be inferred that Soxhlet extracted Azadirachta indica seed may be industrially viable but less suitable for consumption.

Volatiles from the fungal phytopathogen Penicillium aurantiogriseum modulate root metabolism and architecture through proteome resetting.

Volatile compounds (VCs) emitted by the fungal phytopathogen Penicillium aurantiogriseum promote root growth and developmental changes in Arabidopsis. Here we characterised the metabolic and molecular responses of roots to fungal volatiles. Proteomic analyses revealed that these compounds reduce the levels of aquaporins, the iron carrier IRT1 and apoplastic peroxidases. Fungal VCs also increased the levels of enzymes involved in the production of mevalonate (MVA)-derived isoprenoids, nitrogen assimilation and conversion of methionine to ethylene and cyanide. Consistently, fungal VC-treated roots accumulated high levels of hydrogen peroxide (H2 O2 ), MVA-derived cytokinins, ethylene, cyanide and long-distance nitrogen transport amino acids. qRT-PCR analyses showed that many proteins differentially expressed by fungal VCs are encoded by VC non-responsive genes. Expression patterns of hormone reporters and developmental characterisation of mutants provided evidence for the involvement of cyanide scavenging and enhanced auxin, ethylene, cytokinin and H2 O2 signalling in the root architecture changes promoted by fungal VCs. Our findings show that VCs from P. aurantiogriseum modify root metabolism and architecture, and improve nutrient and water use efficiencies through transcriptionally and non-transcriptionally regulated proteome resetting mechanisms. Some of these mechanisms are subject to long-distance regulation by photosynthesis and differ from those triggered by VCs emitted by beneficial microorganisms.

Effects of hydrogen water treatment on antioxidant system of litchi fruit during the pericarp browning

Litchi fruit were exposed to 0.7 PPM hydrogen water (HW) before storage at 25 ± 1 ℃. HW treatment delayed the pericarp browning and maintained the total soluble solids (TSS) of litchi fruit. Then, a total of 25 antioxidant system-related characters were determined to evaluate the effects of HW on antioxidant system during pericarp browning. Compared with control pericarp, the pericarp of HW-treated litchi fruit exhibited higher levels of superoxide radical (O2−·) scavenging activity, glutathione (GSH), monodehydroascorbate reductase (MDHAR), polyphenol oxidase (PPO) and total flavonoids during whole storage, higher levels of hydrogen peroxide (H2O2), catalase (CAT), glutathione disulfide (GSSG), ascorbate oxidase (AAO) and total phenols only on day 1, and higher levels of ascorbate peroxidase (APX), total anthocyanin, glutathione reductase (GR) and glutathione peroxidases (GPX) at later stage of storage. Those HW-induced antioxidant system-related characters might directly or indirectly enhanced the antioxidant capacity and delayed the pericarp browning of litchi.

MicroRNA156 (miR156) Negatively Impacts Mg-Protoporphyrin IX (Mg-Proto IX) Biosynthesis and Its Plastid-Nucleus Retrograde Signaling in Apple.

Plastid-nucleus retrograde signaling (PNRS) play essential roles in regulating nuclear gene expression during plant growth and development. Excessive reactive oxygen species can trigger PNRS. We previously reported that in apple (Malus domestica Borkh.) seedlings, the expression of microRNA156 (miR156) was significantly low in the adult phase, which was accompanied by high levels of hydrogen peroxide (H2O2) accumulation in chloroplasts. However, it was unclear whether adult-phase-specific chloroplast H2O2 may induce PNRS and affect miR156 expression, or miR156 triggers adult phase PNRS during the ontogenesis. In this paper, we examined the relationship between miR156 levels and six PNRS components in juvenile and adult phase leaves from ‘Zisai Pearl’בRed Fuji’ hybrids. We found that PNRS generated by singlet oxygen (1O2), the photosynthetic redox state, methylerythritol cyclodiphosphate (MEcPP), SAL1-3-phosphoadenosine 5-phosphate (PAP) and WHIRLY1 were not involved. The accumulation of Mg-protoporphyrin IX (Mg-Proto IX), the expression of the synthetic genes MdGUN5 and MdGUN6, and Mg-Proto IX PNRS related nuclear genes increased with ontogenesis. These changes were negatively correlated with miR156 expression. Manipulating Mg-Proto IX synthesis with 5-aminolevulinic acid (ALA) or gabaculine did not affect miR156 expression in vitro shoots. In contrast, modulating miR156 expression via MdGGT1 or MdMIR156a6 transgenesis led to changes in Mg-Proto IX contents and the corresponding gene expressions. It was concluded that the Mg-Proto IX PNRS was regulated downstream of miR156 regardless of adult-phase-specific plastid H2O2 accumulation. The findings may facilitate the understanding of the mechanism of ontogenesis in higher plants.

Reactive Oxygen Species, Antioxidant Agents, and DNA Damage in Developing Maize Mitochondria and Plastids.

Maize shoot development progresses from non-pigmented meristematic cells at the base of the leaf to expanded and non-dividing green cells of the leaf blade. This transition is accompanied by the conversion of promitochondria and proplastids to their mature forms and massive fragmentation of both mitochondrial DNA (mtDNA) and plastid DNA (ptDNA), collectively termed organellar DNA (orgDNA). We measured developmental changes in reactive oxygen species (ROS), which at high concentrations can lead to oxidative stress and DNA damage, as well as antioxidant agents and oxidative damage in orgDNA. Our plants were grown under normal, non-stressful conditions. Nonetheless, we found more oxidative damage in orgDNA from leaf than stalk tissues and higher levels of hydrogen peroxide, superoxide, and superoxide dismutase in leaf than stalk tissues and in light-grown compared to dark-grown leaves. In both mitochondria and plastids, activities of the antioxidant enzyme peroxidase were higher in stalk than in leaves and in dark-grown than light-grown leaves. In protoplasts, the amount of the small-molecule antioxidants, glutathione and ascorbic acid, and catalase activity were also higher in the stalk than in leaf tissue. The data suggest that the degree of oxidative stress in the organelles is lower in stalk than leaf and lower in dark than light growth conditions. We speculate that the damaged/fragmented orgDNA in leaves (but not the basal meristem) results from ROS signaling to the nucleus to stop delivering DNA repair proteins to mature organelles producing large amounts of ROS.