Reactive Oxygen Species Metabolism Research Articles

Radiotherapy-induced ferroptosis for cancer treatment.

Ferroptosis is a regulated cell death mechanism controlled by iron, amino acid and reactive oxygen species metabolisms, which is very relevant for cancer therapy. Radiotherapy-induced ferroptosis is critical for tumor suppression and several preclinical studies have demonstrated that the combination of ionizing radiation with small molecules or nano-systems is effective in combating cancer growth and overcoming drug or ionizing radiation resistance. Here, we briefly overview the mechanisms of ferroptosis and the cross-talk existing between the cellular pathways activated by ferroptosis and those induced by radiotherapy. Lastly, we discuss the recently reported combinational studies involving radiotherapy, small molecules as well as nano-systems and report the recent findings achieved in this field for the treatment of tumors.

Salt priming induces low-temperature tolerance in sugar beet via xanthine metabolism

To understand the physiological mechanisms involved in xanthine metabolism during salt priming for improving low-temperature tolerance, salt priming (SP), xanthine dehydrogenase inhibitor (XOI), exogenous allantoin (EA), and back-supplemented EA (XOI + EA) treatments were given and the low-temperature tolerance of sugar beet was tested. Under low-temperature stress, salt priming promoted the growth of sugar beet leaves and increased the maximum quantum efficiency of PS II (Fv/Fm). However, during salt priming, either XOI or EA treatment alone increased the content of reactive oxygen species (ROS), such as superoxide anion and hydrogen peroxide, in the leaves under low-temperature stress. XOI treatment increased allantoinase activity with its gene (BvallB) expression under low-temperature stress. Compared to the XOI treatment, the EA treatment alone and the XOI + EA treatment increased the activities of antioxidant enzymes. At low temperatures, the sucrose content and the activity of key carbohydrate enzymes (AGPase, Cylnv, and FK) were significantly reduced by XOI compared to the changes under salt priming. XOI also stimulated the expression of protein phosphatase 2C and sucrose non-fermenting1-related protein kinase (BvSNRK2). The results of a correlation network analysis showed that BvallB was positively correlated with malondialdehyde, D-Fructose-6-phosphate, and D-Glucose-6-phosphate, and negatively correlated with BvPOX42, BvSNRK2, dehydroascorbate reductase, and catalase. These results suggested that salt-induced xanthine metabolism modulated ROS metabolism, photosynthetic carbon assimilation, and carbohydrate metabolism, thus enhancing low-temperature tolerance in sugar beet. Additionally, xanthine and allantoin were found to play key roles in plant stress resistance.

Cover Caption

AbstractJune Online Cover: Possible mechanism of reactive oxygen species (ROS) metabolism in fruit tissue through the regulation of oxygen concentration in the microenvironment, with Illustrations of avocados in a sealed glass jar and of the microenvironment and inner gas release process; photographs of grapes from the different treatment groups; possible mechanism of the microenvironment delaying fruit senescence through the regulation of oxygen metabolism; and correlation analysis of the microenvironment, from “Antioxidant mechanism of a continuous microenvironment regulation system for prolonging the shelf‐life of red grapes” by Wenqing Hu, Fan Gong, Qingfeng Dong, Yongfeng Kang, Yan Zhou, Boqiang Liu, and Li Li. p. 2496–2511.

Involvement of Nitric Oxide and Melatonin Enhances Cadmium Resistance of Tomato Seedlings through Regulation of the Ascorbate-Glutathione Cycle and ROS Metabolism.

Melatonin (MT) and nitric oxide (NO) act as signaling molecules that can enhance cadmium (Cd) stress resistance in plants. However, little information is available about the relationship between MT and NO during seedling growth under Cd stress. We hypothesize that NO may be involved in how MT responds to Cd stress during seedling growth. The aim of this study is to evaluate the relationship and mechanism of response. The results indicate that different concentrations of Cd inhibit the growth of tomato seedlings. Exogenous MT or NO promotes seedling growth under Cd stress, with a maximal biological response at 100 μM MT or NO. The promotive effects of MT-induced seedling growth under Cd stress are suppressed by NO scavenger 2-4-carboxyphenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (cPTIO), suggesting that NO may be involved in MT-induced seedling growth under Cd stress. MT or NO decreases the content of hydrogen peroxide (H2O2), malonaldehyde (MDA), dehydroascorbic acid (DHA), and oxidized glutathione (GSSG); improves the content of ascorbic acid (AsA) and glutathione (GSH) and the ratios of AsA/DHA and GSH/GSSG; and enhances the activities of glutathione reductase (GR), monodehydroascorbic acid reductase (MDHAR), dehydroascorbic acid reductase (DHAR), ascorbic acid oxidase (AAO), and ascorbate peroxidase (APX) to alleviate oxidative damage. Moreover, the expression of genes associated with the ascorbate-glutathione (AsA-GSH) cycle and reactive oxygen species (ROS) are up-regulated by MT or NO under Cd conditions, including AAO, AAOH, APX1, APX6, DHAR1, DHAR2, MDHAR, and GR. However, NO scavenger cPTIO reverses the positive effects regulated by MT. The results indicate that MT-mediated NO enhances Cd tolerance by regulating AsA-GSH cycle and ROS metabolism.

Differential Responses of Antioxidant Enzymes and Lignin Metabolism in Susceptible and Resistant Sweetpotato Cultivars during Root-Knot Nematode Infection.

Root-knot nematodes (RKN) cause significant damage to sweetpotato plants and cause significant losses in yield and quality. Reactive oxygen species (ROS) play an important role in plant defenses, with levels of ROS-detoxifying antioxidant enzymes tightly regulated during pathogen infection. In this study, ROS metabolism was examined in three RKN-resistant and three RKN-susceptible sweetpotato cultivars. The antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) were assessed, as was lignin-related metabolism. In RKN-infected roots, both resistant and susceptible cultivars increased SOD activity to produce higher levels of hydrogen peroxide (H2O2). However, H2O2 removal by CAT activity differed between cultivars, with susceptible cultivars having higher CAT activity and lower overall H2O2 levels. In addition, the expression of phenylpropanoid-related phenylalanine ammonia-lyase and cinnamyl alcohol dehydrogenase genes, which encode enzymes involved in lignin metabolism, were higher in resistant cultivars, as were total phenolic and lignin contents. Enzyme activities and H2O2 levels were examined during the early (7 days) and late (28 days) phases of infection in representative susceptible and resistant cultivars, revealing contrasting changes in ROS levels and antioxidant responses in the different stages of infection. This study suggests that differences in antioxidant enzyme activities and ROS regulation in resistant and susceptible cultivars might explain reduced RKN infection in resistant cultivars, resulting in smaller RKN populations and overall higher resistance to infection and infestation by RKNs.

Lighting the light reactions of photosynthesis by means of redox-responsive genetically encoded biosensors for photosynthetic intermediates.

Oxygenic photosynthesis involves light and dark phases. In the light phase, photosynthetic electron transport provides reducing power and energy to support the carbon assimilation process. It also contributes signals to defensive, repair, and metabolic pathways critical for plant growth and survival. The redox state of components of the photosynthetic machinery and associated routes determines the extent and direction of plant responses to environmental and developmental stimuli, and therefore, their space- and time-resolved detection in planta becomes critical to understand and engineer plant metabolism. Until recently, studies in living systems have been hampered by the inadequacy of disruptive analytical methods. Genetically encoded indicators based on fluorescent proteins provide new opportunities to illuminate these important issues. We summarize here information about available biosensors designed to monitor the levels and redox state of various components of the light reactions, including NADP(H), glutathione, thioredoxin, and reactive oxygen species. Comparatively few probes have been used in plants, and their application to chloroplasts poses still additional challenges. We discuss advantages and limitations of biosensors based on different principles and propose rationales for the design of novel probes to estimate the NADP(H) and ferredoxin/flavodoxin redox poise, as examples of the exciting questions that could be addressed by further development of these tools. Genetically encoded fluorescent biosensors are remarkable tools to monitor the levels and/or redox state of components of the photosynthetic light reactions and accessory pathways. Reducing equivalents generated at the photosynthetic electron transport chain in the form of NADPH and reduced ferredoxin (FD) are used in central metabolism, regulation, and detoxification of reactive oxygen species (ROS). Redox components of these pathways whose levels and/or redox status have been imaged in plants using biosensors are highlighted in green (NADPH, glutathione, H2O2, thioredoxins). Analytes with available biosensors not tried in plants are shown in pink (NADP+). Finally, redox shuttles with no existing biosensors are circled in light blue. APX, ASC peroxidase; ASC, ascorbate; DHA, dehydroascorbate; DHAR, DHA reductase; FNR, FD-NADP+ reductase; FTR, FD-TRX reductase; GPX, glutathione peroxidase; GR, glutathione reductase; GSH, reduced glutathione; GSSG, oxidized glutathione; MDA, monodehydroascorbate; MDAR, MDA reductase; NTRC, NADPH-TRX reductase C; OAA, oxaloacetate; PRX, peroxiredoxin; PSI, photosystem I; PSII: photosystem II; SOD, superoxide dismutase; TRX, thioredoxin.

Transcriptome analysis reveals the effects of hydrogen gas on NaCl stress in purslane roots

Purslane development may be affected by high salt concentrations. It has been found that H2 enhanced salt-tolerance of plant. The goal of present study was to investigate the impact of H2 on root growth of purslane under salt stress, and to analyze the molecular mechanism of H2 effect on salt tolerance of purslane by using transcriptome method. Under hydroponic conditions, H2 significantly reduced electrolyte leakage and malondialdehyde content, substantially enhance the activities of superoxide dismutase, peroxidase, catalase and ascorbate peroxidase in roots of purslane (‘Tall Green’) under NaCl stress. Transcriptome analysis showed that NaCl stress significantly affected the expression of differential genes in purslane roots. The expression of differential genes was significantly affected by the addition of H2, especially under salt stress. The qRT-PCR analysis was used to confirm the RNA-seq results. GO and KEGG analysis showed that the DEGs were mainly involved in metabolism of reactive oxygen species (ROS), plant hormone signal transduction and UDP-glucosyltransferase activity. At the transcriptional level, ROS accumulation and clearance; auxin, cytokinin, gibberellin, abscisic acid signal transduction; sucrose synthesis and catabolism, and the synthesis of plant cell wall component may play a crucial role in the influence of H2 on the salt tolerance of purslane. These findings have crucial implications for improved understanding the role of H2 in plant salt tolerance and provide specific data for in-depth identification of purslane salt tolerance genes.

A network pharmacological-based study of the mechanism of Liuwei Dihuang pill in the treatment of chronic kidney disease.

Chronic kidney disease (CKD) is a progressive disease that poses a huge economic burden to society. Liuwei Dihuanng pill is an effective treatment for chronic kidney disease, but its treatment mechanism is unclear. The rapid development of network pharmacology has provided new strategies for studying Chinese medicine. The traditional Chinese medicine systems pharmacology database and analysis platform was used to obtain the bioactive components and targets of Liuwei Dihuanng pill. The sources for the CKD-related targets were then obtained from the Genecards, OMIM, TTD, and DisGeNET databases. R was used to identify the intersecting genes for Liuwei Dihuang pill and CKD-related targets. Analysis of protein-protein interactions (PPI) was performed using STRING, and PPI networks and drug-component-target networks were constructed using Cytoscape software. Kyoto encyclopedia of genes and genomes pathway and gene ontology enrichment analyses were performed using R. Finally, molecular docking was performed to determine the binding activity between bioactive components and the targets. After screening and data de-duplication of 74 active components, 209 drug targets, and 14,794 disease targets, a total of 204 drug-disease targets were acquired. Subsequently, a drug-component-target network and PPI network were established. The primary components of Liuwei Dihuang pill included quercetin, stigmasterol, kaempferol, beta-sitosterol, tetrahydroalstonine, kadsurenone, hederagenin, hancinone C, diosgenin, and sitosterol. In addition, JUN, AKT1, TP53, RELA, MAPK1, FOS, TNF, IL6, ESR1, and RXRA were identified as the main targets. Gene ontology function enrichment analysis revealed that these targets were involved in reactive oxygen species metabolic processes, responses to metal ions and to chemical stimuli, G protein-coupled amine receptor activity, and nuclear factor receptor activity. Kyoto encyclopedia of genes and genomes enrichment analysis showed that these targets were involved in the AGE-RAGE signaling pathway, IL-17 signaling pathway, TNF signaling pathway, and so on. Molecular docking results indicated good binding activity between the core targets and core components. The potential mechanism of Liuwei Dihuanng pill in the treatment of CKD was preliminarily discussed in this study, providing a theoretical basis and evidence for further experimental research.

Transcriptome Analysis of the Late-Acting Self-Incompatibility Associated with RNase T2 Family in Camellia oleifera.

The Camellia oil tree (Camellia oleifera Abel.) is an important nonwood forest species in China, and the majority of its cultivars are late-acting self-incompatibility (LSI) types. Although several studies have examined the mechanism of LSI, the process is quite complicated and unclear. In this study, pollen tube growth and fruit setting of two Camellia oil tree cultivars Huashuo (HS) and Huajin (HJ) were investigated after non and self-pollination, and transcriptomic analysis of the ovaries was performed 48 h after self-pollination to identify the potential genes implicated in the LSI of Camellia oil trees. The results showed that the fruit set of HS was significantly higher than that of HJ after self-pollination. Transcriptomic analysis revealed that plant hormone signal transduction, the phosphatidylinositol signaling system, ATP-binding cassette (ABC) transporters, reactive oxygen species (ROS) metabolism, and Ca2+ signaling were mainly contributed in the LSI of reaction of Camellia oil tree. Moreover, nine RNase T2 genes were identified from the transcriptome analysis, which also showed that CoRNase7 participated in the self-incompatibility reaction in HS. Based on phylogenetic analysis, CoRNase6 was closely related to S-RNase from coffee, and CoRNase7 and CoRNase8 were closely related to S-RNase from Camellia sinensis. The 9 RNase T2 genes successfully produced proteins in prokaryotes. Subcellular localization indicated that CoRNase1 and CoRNase5 were cytoplasmic proteins, while CoRNase7 was a plasma membrane protein. These results screened the main metabolic pathways closely related to LSI in Camellia oil tree, and SI signal transduction might be regulated by a large molecular regulatory network. The discovery of T2 RNases provided evidence that Camellia oil tree might be under RNase-based gametophytic self-incompatibility.

A novel effector RipBT contributes to Ralstonia solanacearum virulence on potato.

Ralstonia solanacearum is one of the most destructive plant-pathogenic bacteria, infecting more than 200 plant species, including potato (Solanum tuberosum) and many other solanaceous crops. R. solanacearum has numerous pathogenicity factors, and type III effectors secreted through type III secretion system (T3SS) are key factors to counteract host immunity. Here, we show that RipBT is a novel T3SS-secreted effector by using a cyaA reporter system. Transient expression of RipBT in Nicotiania benthamiana induced strong cell death in a plasma membrane-localization dependent manner. Notably, mutation of RipBT in R. solanacearum showed attenuated virulence on potato, while RipBT transgenic potato plants exhibited enhanced susceptibility to R. solanacearum. Interestingly, transcriptomic analyses suggest that RipBT may interfere with plant reactive oxygen species (ROS) metabolism during the R. solanacearum infection of potato roots. In addition, the expression of RipBT remarkably suppressed the flg22-induced pathogen-associated molecular pattern-triggered immunity responses, such as the ROS burst. Taken together, RipBT acts as a T3SS effector, promoting R. solanacearum infection on potato and presumably disturbing ROS homeostasis.

Identification and validation of ferroptosis-related genes in lipopolysaccharide-induced acute lung injury

BackgroundEmerging evidence reveals the important role of ferroptosis in the pathophysiological process of acute lung injury (ALI). We aimed to identify and validate the potential ferroptosis-related genes of ALI through bioinformatics analysis and experimental validation. MethodsMurine ALI model was established via intratracheal instillation with LPS and confirmed by H&E staining and transmission electronic microscopy (TEM). RNA sequencing (RNA-seq) was used to screen differentially expressed genes (DEGs) between control and ALI model mice. The potential differentially expressed ferroptosis-related genes of ALI were identified using the limma R package. Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, gene set enrichment analysis (GSEA), and protein-protein interactions (PPI) were applied for the differentially expressed ferroptosis-related genes. CIBERSORT tool was used to conduct immune cell infiltration analysis. Finally, protein expressions and RNA expression of ferroptosis DEGs were validated in vivo and in vitro by western blots and RT-qPCR. ResultsAmong 5009 DEGs, a total of 86 differentially expressed ferroptosis-related genes (45 up-regulated genes and 41 down-regulated genes) were identified in the lungs between control and ALI. GSEA analysis showed that the genes enriched were mainly involved in response to molecule of bacterial origin and fatty acid metabolic process. The GO and KEGG enrichment analysis indicated that the top 40 ferroptosis DEGs were mainly enriched in reactive oxygen species metabolic process, HIF-1signaling pathway, lipid and atherosclerosis, and ferroptosis. The PPI results and Spearman correlation analysis suggested that these ferroptosis-related genes interacted with each other. Immune infiltration analysis confirmed that ferroptosis DEGs were closely related to immune response. Consistent with the RNA-seq data, the western blot and RT-qPCR unveiled increased mRNA expressions of Cxcl2, Il-6, Il-1β, and Tnfα, and protein expressions of FTH1, TLR4 as well as decreased ACSL3 in LPS-induced ALI. In vitro, the upregulated mRNA levels of CXCL2, IL-6, SLC2A1, FTH1, TNFAIP3, and downregulated NQO1 and CAV1 in LPS-stimulated BEAS-2B and A549 cells were verified. ConclusionWe identified 86 potential ferroptosis-related genes of LPS-induced ALI through RNA-seq. Several pivotal ferroptosis-related genes involved in lipid metabolism and iron metabolism were implicated in ALI. This study may be helpful to expand our understanding of ALI and provide some potential targets to counteract ferroptosis in ALI.

Bergapten inhibits NLRP3 inflammasome activation and pyroptosis via promoting mitophagy.

Inhibition of NLRP3 inflammasome activation produces potent therapeutic effects in a wide array of inflammatory diseases. Bergapten (BeG), a furocoumarin phytohormone present in many herbal medicines and fruits, exibits anti-inflammatory activity. In this study we characterized the therapeutic potential of BeG against bacterial infection and inflammation-related disorders, and elucidated the underlying mechanisms. We showed that pre-treatment with BeG (20 μM) effectively inhibited NLRP3 inflammasome activation in both lipopolysaccharides (LPS)-primed J774A.1 cells and bone marrow-derived macrophages (BMDMs), evidenced by attenuated cleaved caspase-1 and mature IL-1β release, as well as reduced ASC speck formation and subsequent gasdermin D (GSDMD)-mediated pyroptosis. Transcriptome analysis revealed that BeG regulated the expression of genes involved in mitochondrial and reactive oxygen species (ROS) metabolism in BMDMs. Moreover, BeG treatment reversed the diminished mitochondrial activity and ROS production after NLRP3 activation, and elevated the expression of LC3-II and enhanced the co-localization of LC3 with mitochondria. Treatment with 3-methyladenine (3-MA, 5 mM) reversed the inhibitory effects of BeG on IL-1β, cleaved caspase-1 and LDH release, GSDMD-N formation as well as ROS production. In mouse model of Escherichia coli-induced sepsis and mouse model of Citrobacter rodentium-induced intestinal inflammation, pre-treatment with BeG (50 mg/kg) significantly ameliorated tissue inflammation and injury. In conclusion, BeG inhibits NLRP3 inflammasome activation and pyroptosis by promoting mitophagy and maintaining mitochondrial homeostasis. These results suggest BeG as a promising drug candidate for the treatment of bacterial infection and inflammation-related disorders.

Ferulic acid enhanced resistance against blue mold of Malus domestica by regulating reactive oxygen species and phenylpropanoid metabolism

Postharvest decay of fruit caused by phytopathogen often causes tremendous losses during logistics. Ferulic acid is a kind of phenolic acid which has potential antimicrobial ability and initiates defense responses. This study performed to evaluate the induced resistance of ferulic acid against blue mold of apples (cv. Qiujin) as well as the mechanism involved in its action. Results suggested that 1.0 g L−1 ferulic acid remarkably reduced lesion diameter of Penicillium expansum-inoculated apples and colony diameter in vitro. The activities and gene expressions of catalase, superoxide dismutase, peroxidase, dehydroascorbate reductase, glutathione reductase, monodehydroascorbate reductase, ascorbate peroxidase, and nicotinamide adenine dinucleotide phosphate oxidase were distinctly improved by ferulic acid immersion. Ferulic acid also promoted ascorbic acid and reduced glutathione contents, whereas decreased hydrogen peroxide and malondialdehyde contents in apples. Moreover, ferulic acid enhanced the gene expressions and activities of cinnamyl alcohol dehydrogenase, 4-coumarate coenzyme A ligase, cinnamate-4-hydroxylase, and phenylalanine ammonia-lyase, resulting in caffeic acid, flavonoids, lignin, total phenolics, and p-coumaric acid accumulation in apples. Taken together, ferulic acid could activate the activities and gene expressions of the key enzymes in phenylpropanoid pathway as well as reactive oxygen species metabolism to promote the synthesis of antioxidants and secondary metabolites, thereby increasing disease resistance of apples.

Transcriptome profiling of Nephelium lappaceum: Sodium nitroprusside-induced delays of postharvest browning.

Sodium nitroprusside (SNP), as a nitric oxide donor, is widely used in postharvest fruit physiology and metabolism. Our previous study has indicated that SNP plays a crucial role in postharvest browning control of rambutan, but the molecular mechanism underlying this process is still unclear. In this research, we investigated the gene expression and function of postharvest rambutan in response to SNP during browning. We found 7336 differentially expressed genes (DEGs), among which 2206 were upregulated and 5130 were downregulated. Gene Ontology (GO) enrichment as well as Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed, and the real-time quantitative PCR (qPCR) data were consistent with transcriptome data. The DEGs relevant to rambutan pericarp browning were mainly involved in anthocyanin biosynthesis, phenolic oxidation, reactive oxygen species (ROS) production, and energy supply. It was shown that SNP regulated the synthesis and degradation of anthocyanins, accumulation of phenols, level of ROS and energy metabolism to suppress the postharvest browning of rambutan. Also, one WRKY transcription factor involved in ROS metabolism was observed to be differentially regulated. These findings add to our insights into the molecular mechanisms of the SNP-induced browning delays of rambutan, which has implications for subsequent studies on molecular mechanisms of fruit browning.

The critical role of copper homeostasis during the immune response

Abstract Copper homeostasis is tightly regulated in living organisms, as copper is essential to numerous cellular processes but toxic in excess. Dysregulated copper homeostasis is associated with numerous human diseases, including genetic diseases of copper transport, cancer, autoimmune disease, and infection. Although patients with mutations in copper transporters have an increased susceptibility to infection, little is known about the changes in immune function in these diseases. Further, altering copper availability has shown promise in treating either autoimmunity or infection, but the mechanisms underlying this effect are unresolved. We examined serum copper levels in human samples during acute influenza or SARS-CoV-2 virus infections, revealing unique virus-dependent copper dynamics. As copper is required for important immune cell processes such as metabolism, signaling and maintenance of reactive oxygen species, we next investigated the effect of modulating copper levels on T cell activation. We found that copper chelation impaired T cell proliferation, reduced expression of T cell activation markers, and altered gene expression following stimulation. To determine the impact of copper levels during the immune response in vivo, we then treated mice with copper chelation prior to and during an influenza A virus infection. Notably, mice treated with copper chelation prior to infection had altered serum copper dynamics, greater weight loss, and more flu-specific T cells compared to untreated mice. Overall, our results demonstrate the importance of copper homeostasis during the immune response. Ongoing experiments will investigate the mechanisms underlying this role in immune function during disease. Supported by ALSAC and NIH (5T32AI106700).

Reactive Oxygen Species Metabolism Modulation on the Quality of Apple Fruits Inoculated with Penicillium expansum under Different Ambient pHs

Apple blue mold is a significant postharvest disease caused by Penicillium expansum. pH modification in colonized tissues leads to the production of organic substances, the modulation of enzymes, and then increases fungal pathogenicity. This study evaluated Penicillium expansum-inoculated apple fruits’ quality responding to pH treatments ranging from 2.5 to 8.5 and analyzed the reactive oxygen species (ROS) metabolism modulation in inoculated apple fruits at the same pH. The results showed that the fruit quality of the firmness, total soluble solids, and titratable acid displayed a quick loss at pHs 5.0 and 7.0, compared with 2.5 and 8.5. Similarly, higher disease incidence was observed at pHs 5.0 and 7.0. Apple fruits infected with P. expansum at pHs 2.5 and 8.5 had less content of O2•−, H2O2, and malondialdehyde (MDA); lower enzymatic activity of NADPH oxidase (NOX); and greater cell membrane integrity than those at pHs 5.0 and 7.0. The analysis of the antioxidant enzymatic activities showed upregulation of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), and glutathione reductase (GR) at pHs 2.5 and 8.5 compared with those at pHs 5.0 and 7.0. Similar trends were shown in ascorbic acid and glutathione. These results support the hypothesis that inoculated apple fruits at pHs 2.5 and 8.5 improve resistance to P. expansum by modulating ROS metabolism, compared with pHs 5.0 and 7.0.

The translocase of the inner mitochondrial membrane 22-2 is required for mitochondrial membrane function during Arabidopsis seed development.

The carrier translocase (also known as translocase of the inner membrane 22; TIM22 complex) is an important component of the mitochondrial protein import apparatus. However, the biological functions of AtTIM22-2 in Arabidopsis remain poorly defined. Here, we report studies on two tim22-2 mutants that exhibit defects in embryo and endosperm development, leading to seed abortion. AtTIM22-2, which was localized in mitochondria, was widely expressed in embryos and in various seedling organs. Loss of AtTIM22-2 function resulted in irregular mitochondrial cristae, decreased respiratory activity, and a lower membrane potential, together with changes in gene expression and enzyme activity related to reactive oxygen species (ROS) metabolism, leading to increased accumulation of ROS in the embryo. The levels of transcripts encoding mitochondrial protein import components were also altered in the tim22-2 mutants. Furthermore, mass spectrometry, bimolecular fluorescence complementation and co-immunoprecipitation assays revealed that AtTIM22-2 interacted with AtTIM23-2, AtB14.7 (a member of Arabidopsis OEP16 family encoded by At2G42210), and AT5G27395 (mitochondrial inner membrane translocase complex, subunit TIM44-related protein). Taken together, these results demonstrate that AtTIM22-2 is essential for maintaining mitochondrial membrane functions during seed development. These findings lay the foundations for a new model of the composition and functions of the TIM22 complex in higher plants.

Genomic capacities for Reactive Oxygen Species metabolism across marine phytoplankton.

Marine phytoplankton produce and scavenge Reactive Oxygen Species, to support cellular processes, while limiting damaging reactions. Some prokaryotic picophytoplankton have, however, lost all genes encoding scavenging of hydrogen peroxide. Such losses of metabolic function can only apply to Reactive Oxygen Species which potentially traverse the cell membrane outwards, before provoking damaging intracellular reactions. We hypothesized that cell radius influences which elements of Reactive Oxygen Species metabolism are partially or fully dispensable from a cell. We therefore investigated genomes and transcriptomes from diverse marine eukaryotic phytoplankton, ranging from 0.4 to 44 μm radius, to analyze the genomic allocations encoding enzymes metabolizing Reactive Oxygen Species. Superoxide has high reactivity, short lifetimes and limited membrane permeability. Genes encoding superoxide scavenging are ubiquitous across phytoplankton, but the fractional gene allocation decreased with increasing cell radius, consistent with a nearly fixed set of core genes for scavenging superoxide pools. Hydrogen peroxide has lower reactivity, longer intracellular and extracellular lifetimes and readily crosses cell membranes. Genomic allocations to both hydrogen peroxide production and scavenging decrease with increasing cell radius. Nitric Oxide has low reactivity, long intracellular and extracellular lifetimes and readily crosses cell membranes. Neither Nitric Oxide production nor scavenging genomic allocations changed with increasing cell radius. Many taxa, however, lack the genomic capacity for nitric oxide production or scavenging. The probability of presence of capacity to produce nitric oxide decreases with increasing cell size, and is influenced by flagella and colony formation. In contrast, the probability of presence of capacity to scavenge nitric oxide increases with increasing cell size, and is again influenced by flagella and colony formation.

Effect of Nisin on the Quality and Antioxidant Activity of Fresh-Cut Pumpkins (Cucurbita moschata Duch.)

Fresh-cut pumpkins refer to fresh pumpkin that has been graded, cleaned, peeled, sliced, preserved, and packaged. It has the qualities of freshness, nutrition, convenience, and being 100% edible. However, mechanical damages during the cutting processing can accelerate the quality deterioration, aging, and loss of nutritional values of fresh-cut pumpkins. Nisin, a natural preservative, has been widely used in fruits and vegetables with good preservation effects. To investigate the effect of different concentrations (0, 0.2, 0.4, and 0.6 g/L) of nisin on the quality of fresh-cut pumpkins, the critical indexes involved in weight loss, firmness, color, respiration intensity, reactive oxygen species (ROS) metabolism, ascorbate (AsA)—glutathione (GSH) cycle, and antioxidant capacity were monitored for fresh-cut pumpkins during storage at 4 °C for 10 days. The results showed that 0.4 g/L nisin was the best preservation concentration. Compared with 0 g/L nisin, 0.4 g/L nisin reduced the weight loss rate and whitening rate of fresh-cut pumpkins by 13.53% and 13.61%, inhibited respiration rate by 45.83%, and maintained hardness by 1.18 times. Meanwhile, 0.4 g/L nisin increased the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) and maintained higher contents of GSH and AsA. It prevented the rapid increase in ROS levels by improving antioxidant capacity, including DPPH, ABTS free radical scavenging rate, and T-AOC (total antioxidant capacity). The collected results showed that nisin has an obvious influence on the quality by regulating physiological and antioxidant activity metabolism. It is envisaged that the combination of nisin and physical and chemical preservation technology will further enhance the quality of fresh-cut pumpkins during storage in the future.

A risk assessment on Zostera chilensis, the last relict of marine angiosperms in the South-East Pacific Ocean, due to the development of the desalination industry in Chile

Seagrasses, which are considered among the most ecologically valuable and endangered coastal ecosystems, have a narrowly limited distribution in the south-east Pacific, where Zostera chilensis is the only remaining relict. Due to water scarcity, desalination industry has grown in the last decades in the central-north coasts of Chile, which may be relevant to address in terms of potential impacts on benthic communities due to their associated high-salinity brine discharges to subtidal ecosystems. In this work, we assessed ecophysiological and cellular responses to desalination-extrapolable hypersalinity conditions on Z. chilensis. Mesocosms experiments were performed for 10 days, where plants were exposed to 3 different salinity treatments: 34 psu (control), 37 psu and 40 psu. Photosynthetic performance, H2O2 accumulation, and ascorbate content (reduced and oxidized) were measured, as well as relative gene expression of enzymes related to osmotic regulation and oxidative stress; these, at 1, 3, 6 and 10 days. Z. chilensis showed a decrease in photosynthetic parameters such as electron transport rate (ETRmax) and saturation irradiance (EkETR) under hypersalinity treatments, while non-photochemical quenching (NPQmax) presented an initial increment and a subsequent decline at 40 psu. H2O2 levels increased with hypersalinity, while ascorbate and dehydroascorbate only increased under 37 psu, although decreased along the experimental period. Increased salinities also triggered the expression of genes related to ion transport and osmolyte syntheses, but salinity-dependent up-regulated genes were mostly those related to the reactive oxygen species metabolism. The relict seagrass Z. chilensis has shown to withstand increased salinities that may be extrapolable to desalination effects in the short-term. As the latter is not fully clear in the long-term, and considering the restricted distribution and ecological importance, direct brine discharges to Z. chilensis meadows may not be recommended.