Reactive Oxygen Species Scavenging Enzymes Research Articles

Flagellin, a plant-defense-activating protein identified from Xanthomonas axonopodis pv. Dieffenbachiae invokes defense response in tobacco

BackgroundSecretome analysis is a valuable tool to study host-pathogen protein interactions and to identify new proteins that are important for plant health. Microbial signatures elicit defense responses in plants, and by that, the plant immune system gets triggered prior to pathogen infection. Functional properties of secretory proteins from Xanthomonas axonopodis pv. dieffenbachiae (Xad1) involved in priming plant immunity was evaluated.ResultsIn this study, the secretome of Xad1 was analyzed under host plant extract-induced conditions, and mass spectroscopic analysis of differentially expressed protein was identified as plant-defense-activating protein viz., flagellin C (FliC). The flagellin and Flg22 peptides both elicited hypersensitive reaction (HR) in non-host tobacco, activated reactive oxygen species (ROS) scavenging enzymes, and increased pathogenesis-related (PR) gene expression viz., NPR1, PR1, and down-regulation of PR2 (β-1,3-glucanase). Protein docking studies revealed the Flg22 epitope of Xad1, a 22 amino acid peptide region in FliC that recognizes plant receptor FLS2 to initiate downstream defense signaling.ConclusionThe flagellin or the Flg22 peptide from Xad1 was efficient in eliciting an HR in tobacco via salicylic acid (SA)-mediated defense signaling that subsequently triggers systemic immune response epigenetically. The insights from this study can be used for the development of bio-based products (small PAMPs) for plant immunity and health.

ROSes-FINDER: a multi-task deep learning framework for accurate prediction of microorganism reactive oxygen species scavenging enzymes.

Reactive oxygen species (ROS) are highly reactive molecules that play important roles in microbial biological processes. However, excessive accumulation of ROS can lead to oxidative stress and cellular damage. Microorganism have evolved a diverse suite of enzymes to mitigate the harmful effects of ROS. Accurate prediction of ROS scavenging enzymes classes (ROSes) is crucial for understanding the mechanisms of oxidative stress and developing strategies to combat related diseases. Nevertheless, the existing approaches for categorizing ROS-related proteins exhibit certain drawbacks with regards to their precision and inclusiveness. To address this, we propose a new multi-task deep learning framework called ROSes-FINDER. This framework integrates three component methods using a voting-based approach to predict multiple ROSes properties simultaneously. It can identify whether a given protein sequence is a ROSes and determine its type. The three component methods used in the framework are ROSes-CNN, which extracts raw sequence encoding features, ROSes-NN, which predicts protein functions based on sequence information, and ROSes-XGBoost, which performs functional classification using ensemble machine learning. Comprehensive experiments demonstrate the superior performance and robustness of our method. ROSes-FINDER is freely available at https://github.com/alienn233/ROSes-Finder for predicting ROSes classes.

Allantoin: A Potential Compound for the Mitigation of Adverse Effects of Abiotic Stresses in Plants.

Stress-induced alterations vary with the species of plants, the intensity and duration of the exposure, and stressors availability in nature or soil. Purine catabolism acts as an inherent defensive mechanism against various abiotic stresses and plays a pivotal role in the stress acclimatisation of plants. The intermediate metabolite of purine catabolism, allantoin, compensates for soil nitrogen deficiency due to the low carbon/nitrogen ratio, thereby maintaining nitrogen homeostasis and supporting plant growth and development. Allantoin accounts for 90% of the total nitrogenous compound in legumes, while it contributes only 15% in non-leguminous plants. Moreover, studies on a variety of plant species have reported the differential accumulation of allantoin in response to abiotic stresses, endowing allantoin as a stress modulator. Allantoin functions as signalling molecule to stimulate stress-responsive genes (P5CS; pyrroline-5-carboxylase synthase) and ROS (reactive oxygen species) scavenging enzymes (antioxidant). Moreover, it regulates cross-talk between the abscisic acid and jasmonic acid pathway, and maintains ion homeostasis by increasing the accumulation of putrescine and/or spermine, consequently enhancing the tolerance against stress conditions. Further, key enzymes of purine catabolism (xanthine dehydrogenase and allantoinase) have also been explored by constructing various knockdown/knockout mutant lines to decipher their impact on ROS-mediated oxidative injury in plants. Thus, it is established that allantoin serves as a regulatory signalling metabolite in stress protection, and therefore a lower accumulation of allantoin also reduces plant stress tolerance mechanisms. This review gives an account of metabolic regulation and the possible contribution of allantoin as a photo protectant, osmoprotectant, and nitrogen recycler to reduce abiotic-stress-induced impacts on plants.

Methyl salicylate-accelerated cellular NO production improves mitochondrial and cytosolic antioxidants in preserving ‘Sucrier’ banana peel spotting during storage

The mitochondria, serving as the central organelle, play a pivotal role in driving cellular processes and orchestrating senescence in plant. Additionally, they are implicated in the occurrence of physiological disorders in harvested crops. This study aimed to elucidate the connection between mitochondrial activity and the development of senescent spotting and uncover the supplementary function of MeSA immersion in facilitating the generation of nitric oxide (NO), which in turn activates the expression of mitochondrial and cytosolic reactive oxygen species (ROS) scavenging enzymes, thereby enhancing their efficiency in neutralizing ROS. The findings of this study revealed a compelling association between diminished mitochondrial activity and the development of peel spotting. Notably, a decline in mitochondrial redox status on day 3 to day 6, as evidenced by triphenyltrtrazolium chloride reduction, coupled with reduction in the expression of antioxidant genes, became increasingly prominent as the severity of spotting lesions intensified, particularly in the vicinity of the affected areas. Intriguingly, treatment with MeSA exhibited a significant elevation in NO accumulation during the initial 3 d coinciding with the upregulation of mitochondrial antioxidant gene expressions. The augmented expression of these genes facilitated the preservation of mitochondrial redox capacity, consequently leading to a reduction in oxidative damage, which corresponded remarkably well with the decrease in peel spotting. Hence, our study highlights the role of MeSA upregulated NO-mediated antioxidative genes, which, in turn, assume a pivotal function in safeguarding mitochondrial integrity and mitigating the occurrence of peel spotting in ‘Sucrier’ bananas.

Hydrogen Sulfide Attenuates Mesenchymal Stem Cell Aging Progress via the Calcineurin-NFAT Signaling Pathway.

Aging is a gradual process that is coupled with a decline in the regenerative capacity of stem cells and a subsequent reduction in tissue function and repair. Hydrogen sulfide (H2S) plays an important role in maintaining the function of stem cells. The present study aimed to investigate the role of H2S in mesenchymal stem cell aging and the underlying mechanism and to provide novel insights into stem cell therapies in elderly people. Bone marrow mesenchymal stem cells (BMMSCs) were isolated from young mice (2 months) and from old mice (12 months). Senescence-associated β-galactosidase (SA-β-Gal) activity, reactive oxygen species (ROS) production, ROS scavenging enzymes, and the expression of cell-cycle-related genes were compared between those young and old BMMSCs. The expression of H2S-producing enzymes and the production of H2S in BMMSCs were examined. In vitro osteogenic differentiation and cell senescence were analyzed in young and old BMMSCs before and after H2S treatment. The underlying mechanism was investigated using calcineurin and NFAT1 inhibitors or a Foxp3 siRNA. Bone volume/tissue volume (BV/TV) of femurs in mice was examined using micro-CT with or without systemic injection of an H2S donor. Here, we found that H2S levels in BMMSCs declined with age. When the generation of H2S was blocked with the CBS inhibitor hydroxylamine and the CSE inhibitor dl-propargylglycine, BMMSCs underwent senescence. The elevation of H2S levels rescued BMMSC function in vitro and prevented bone loss in vivo. Mechanistically, H2S represses cell aging via the calcineurin-NFAT1 signaling pathway.

Divergent Cross-Adaptation of Herbicide-Treated Wheat and Triticale Affected by Drought or Waterlogging.

Widely used agrochemicals that do not exert negative effects on crops and selectively target weeds could influence plant resilience under unfavorable conditions. The cross-adaptation of wheat (Triticum aestivum L.) and triticale (×Triticosecale Wittm.) exposed to two environmental abiotic stressors (drought and waterlogging) was evaluated after treatment with a selective herbicide (Serrate®, Syngenta). The ambivalent effects of the herbicide on the two studied crops were particularly distinct in waterlogged plants, showing a significant reduction in wheat growth and better performance of triticale individuals exposed to the same combined treatment. Histochemical staining for the detection of reactive oxygen species (ROS) confirmed that the herbicide treatment increased the accumulation of superoxide anion in the flooded wheat plants, and this effect persisted in the younger leaves of the recovered individuals. Comparative transcript profiling of ROS scavenging enzymes (superoxide dismutase, peroxidase, glutathione reductase, and catalase) in stressed and recovered plants revealed crop-specific variations resulting from the unfavorable water regimes in combination with the herbicide treatment. Short-term dehydration was relatively well tolerated by the hybrid crop triticale and this aligned with the considerable upregulation of genes for L-Proline biosynthesis. Its drought resilience was diminished by herbicide application, as evidenced by increased ROS accumulation after prolonged water deprivation.

Variability and heterosis of seed vigour traits in hybrid rice (Oryza sativa L.)

The cultivars with high seed vigor are desirable for farmers to establish optimum plant stand under sub-optimal field conditions. The basic physiological processes are required to understand the hybrid vigor phenomenon, particularly for seed vigor and associated characteristics. Limited information on seed vigor in rice hybrids and their parental lines in relation to physical, physiological and biochemical characters are available. Three released rice hybrids and their respective female (A), maintainer (B) and restorer/male (R) lines were assessed for physical, physiological and biochemical traits related to seed vigor. Significant differences were found among the three rice hybrids and its parental lines for 100 seed weight, weight of kernel, husk, endosperm and embryo. Germination percentage, seed vigor index-I and seed vigor index-II were highly significant among female and male lines and hybrids in fresh seed because of potential heterosis in hybrids. Higher activity of reactive oxygen species (ROS) scavenging enzymes such as catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) in hybrids was estimated as compared to that of their parental lines. The study showed the presence of the genetic variability, phenotypic, genotypic and environmental coefficient of variation, heritability, genetic advance and heterosis for seed vigor traits in three rice hybrids and its parental lines. A favorable combination of seed vigor traits would be useful to develop desired variety and hybrids with superior seed vigour for realizing higher grain yield and better adaptability in any condition.

Tardigrade proteins: molecular tools in the phenomenon of anhydrobiosis

Abstract Tardigrada (water bears) are a group of small invertebrates known for their ability to survive extreme desiccation by entering an anhydrobiotic state. This state is characterized by the loss of ≤ 98% of body water, resulting in body shrinkage and almost complete cessation of metabolism. In anhydrobiosis, multiple mechanical and biochemical stress effects, such as protein denaturation and aggregation, DNA damage, formation of reactive oxygen species, membrane disintegration and degeneration of cellular compartments, are counteracted by various proteins. Although multiple studies of individual proteins and tardigrade proteomes have been conducted, our knowledge of tardigrade anhydrobiosis-related proteins has not yet been reviewed. To fill this gap, based on available papers, we present a review of all tardigrade proteins known to contribute to desiccation survival, including intrinsically disordered proteins, heat shock proteins and reactive oxygen species-scavenging enzymes. Although proteins are the major anhydrobiosis-related factors in tardigrades, many questions regarding the role of the tardigrade proteome in anhydrobiosis remain unanswered.

Protective Activity of Rhizobium leguminosarum bv. viciae Strain 33504-Mat209 against Alfalfa Mosaic Virus Infection in Faba Bean Plants.

The application of Rhizobium spp., nitrogen-fixing plant growth-promoting rhizobacteria, as biocontrol agents to enhance systemic disease resistance against plant viral infections is a promising approach towards achieving sustainable and eco-friendly agriculture. However, their potential as antivirals and biocontrol agents is less studied. Herein, the capability of Rhizobium leguminosarum bv. viciae strain 33504-Mat209 was evaluated to promote plant growth and enhance faba bean systemic resistance against alfalfa mosaic virus (AMV) infection. Under greenhouse conditions, the soil inoculation with 3504-Mat209 resulted in notable improvements in growth and an increase in chlorophyll content. This led to a marked decrease in the disease incidence, severity, and viral accumulation level by 48, 74, and 87%, respectively. The protective effect of 33504-Mat209 was linked to significant decreases in non-enzymatic oxidative stress indicators, specifically H2O2 and MDA. Additionally, there were significant increases in the activity of reactive oxygen species scavenging enzymes, such as peroxidase (POX) and polyphenol oxidase (PPO), compared to the virus treatment. The elevated transcript levels of polyphenolic pathway genes (C4H, HCT, C3H, and CHS) and pathogenesis-related protein-1 were also observed. Out of 18 detected compounds, HPLC analysis revealed that 33504-Mat209-treated plants increased the accumulation of several compounds, such as gallic acid, chlorogenic acid, catechin, pyrocatechol, daidzein, quercetin, and cinnamic acid. Therefore, the ability of 33504-Mat209 to promote plant growth and induce systemic resistance against AMV infection has implications for utilizing 33504-Mat209 as a fertilizer and biocontrol agent. This could potentially introduce a new strategy for safeguarding crops, promoting sustainability, and ensuring environmental safety in the agricultural sector. As far as we know, this is the first study of biological control of AMV mediated by Rhizobium spp. in faba bean plants.

Effects of ROS pathway inhibitors and NADH and FADH2 linked substrates on mitochondrial bioenergetics and ROS emission in the heart and kidney cortex and outer medulla

Mitochondria are major sources of reactive oxygen species (ROS), which play important roles in both physiological and pathological processes. However, the specific contributions of different ROS production and scavenging components in the mitochondria of metabolically active tissues such as heart and kidney cortex and outer medulla (OM) are not well understood. Therefore, the goal of this study was to determine contributions of different ROS production and scavenging components and provide detailed comparisons of mitochondrial respiration, bioenergetics, ROS emission between the heart and kidney cortex and OM using tissues obtained from the same Sprague-Dawley rat under identical conditions and perturbations. Specifically, data were obtained using both NADH-linked substrate pyruvate + malate and FADH2-linked substrate succinate followed by additions of inhibitors of different components of the electron transport chain (ETC) and oxidative phosphorylation (OxPhos) and other ROS production and scavenging systems. Currently, there is limited data available for the mitochondria of kidney cortex and OM, the two major energy-consuming tissues in the body only next to the heart, and scarce quantitative information on the interplay between mitochondrial ROS production and scavenging systems in the three tissues. The findings from this study demonstrate significant differences in mitochondrial respiratory and bioenergetic functions and ROS emission among the three tissues. The results quantify the rates of ROS production from different complexes of the ETC, identify the complexes responsible for variations in mitochondrial membrane depolarization and regulations of ROS production, and quantify the contributions of ROS scavenging enzymes towards overall mitochondrial ROS emission. These findings advance our fundamental knowledge of tissue-specific and substrate-dependent mitochondrial respiratory and bioenergetic functions and ROS emission. This is important given the critical role that excess ROS production, oxidative stress, and mitochondrial dysfunction in the heart and kidney cortex and OM play in the pathogenesis of cardiovascular and renal diseases, including salt-sensitive hypertension.

Physiological and Genetic Modifications Induced by Plant-Growth-Promoting Rhizobacteria (PGPR) in Tomato Plants under Moderate Water Stress.

Physiological, metabolic, and genetic changes produced by two plant growth promoting rhizobacteria (PGPR) Pseudomonas sp. (internal code of the laboratory: N 5.12 and N 21.24) inoculated in tomato plants subjected to moderate water stress (10% polyethylene glycol-6000; PEG) were studied. Photosynthesis efficiency, photosynthetic pigments, compatible osmolytes, reactive oxygen species (ROS) scavenging enzymes activities, oxidative stress level and expression of genes related to abscisic acid synthesis (ABA; 9-cis-epoxycarotenoid dioxygenase NCDE1 gene), proline synthesis (Pyrroline-5-carboxylate synthase P5CS gene), and plasma membrane ATPase (PM ATPase gene) were measured. Photosynthetic efficiency was compromised by PEG, but bacterial-inoculated plants reversed the effects: while N5.12 increased carbon fixation (37.5%) maintaining transpiration, N21.24 increased both (14.2% and 31%), negatively affecting stomatal closure, despite the enhanced expression of NCDE1 and plasma membrane ATPase genes, evidencing the activation of different adaptive mechanisms. Among all parameters evaluated, photosynthetic pigments and antioxidant enzymes guaiacol peroxidase (GPX) and ascorbate peroxidase (APX) responded differently to both strains. N 5.12 increased photosynthetic pigments (70% chlorophyll a, 69% chlorophyll b, and 65% carotenoids), proline (33%), glycine betaine (4.3%), and phenolic compounds (21.5%) to a greater extent, thereby decreasing oxidative stress (12.5% in Malondialdehyde, MDA). Both bacteria have highly beneficial effects on tomato plants subjected to moderate water stress, improving their physiological state. The use of these bacteria in agricultural production systems could reduce the amount of water for agricultural irrigation without having a negative impact on food production.

Selective oxidative protection leads to tissue topological changes orchestrated by macrophage during ulcerative colitis

Ulcerative colitis is a chronic inflammatory bowel disorder with cellular heterogeneity. To understand the composition and spatial changes of the ulcerative colitis ecosystem, here we use imaging mass cytometry and single-cell RNA sequencing to depict the single-cell landscape of the human colon ecosystem. We find tissue topological changes featured with macrophage disappearance reaction in the ulcerative colitis region, occurring only for tissue-resident macrophages. Reactive oxygen species levels are higher in the ulcerative colitis region, but reactive oxygen species scavenging enzyme SOD2 is barely detected in resident macrophages, resulting in distinct reactive oxygen species vulnerability for inflammatory macrophages and resident macrophages. Inflammatory macrophages replace resident macrophages and cause a spatial shift of TNF production during ulcerative colitis via a cytokine production network formed with T and B cells. Our study suggests components of a mechanism for the observed macrophage disappearance reaction of resident macrophages, providing mechanistic hints for macrophage disappearance reaction in other inflammation or infection situations.

Effects of Reactive Oxygen Levels on Chilling Injury and Storability in 21 Apricot Varieties from Different Production Areas in China.

The key factors for resistance to chilling injury in apricot fruits were obtained by analyzing the low-temperature storage characteristics of 21 varieties of apricot fruits in the main producing areas of China. Twenty-one varieties of apricots from different production areas in China were stored at 0 °C for 50 d and then shelved at 25 °C. The storage quality, chilling injury, reactive oxygen species (ROS), antioxidant ability, and contents of bioactive substances of the apricots were measured and analyzed. The results showed that the 21 varieties of apricot fruits could be divided into two categories according to tolerance during low-temperature storage, where there was chilling tolerance and lack of chilling tolerance. Eleven varieties of apricots, of which Xiangbai and Yunbai are representative, suffered from severe chilling injury after cold storage and shelf life. After 50 d of storage at 0 °C, the levels of superoxide anions and hydrogen peroxide accumulated in the 11 varieties of apricots with a lack of chilling tolerance during storage were significantly higher than those in the remaining 10 varieties of apricots with chilling tolerance. In addition, the activities of ROS scavenging enzymes, represented by superoxide dismutase, catalase and peroxidase, were significantly decreased in 11 varieties of apricots with a lack of chilling tolerance during storage. The contents of bioactive substances with ROS scavenging ability, represented by ascorbic acid, total phenols, carotenoids, and total flavonoids, also significantly decreased. The 10 varieties of apricots, of which Akeximixi and Suanmao are representative, were less affected by chilling injury because the production and removal of ROS were maintained at normal levels, avoiding the damaging effects of ROS accumulation in the fruit. In addition, the 10 apricot varieties with chilling tolerance during storage had higher sugar and acid contents after harvest. This could supply energy for physiological metabolism during cold storage and provide carbon skeletons for secondary metabolism, thus enhancing the chilling tolerance of the fruits. Based on the results of cluster analysis combined with the geographical distribution of the 21 fruit varieties, it was found that apricot varieties with chilling tolerance during storage were all from the northwestern region of China where diurnal temperature differences and rapid climate changes occur. In conclusion, maintaining the balance of ROS production and removal in apricots during cold storage is a key factor to enhance the storage tolerance of apricots. Moreover, apricots with higher initial glycolic acid and bioactive substance contents are less susceptible to chilling injury.

ROS accumulation-induced tapetal PCD timing changes leads to microspore abortion in cotton CMS lines

BackgroundCytoplasmic male sterility (CMS) is the basis of heterosis exploitation. CMS has been used to hybrid production in cotton, but its molecular mechanism remains unclear. CMS is associated with advanced or delayed tapetal programmed cell death (PCD), and reactive oxygen species (ROS) may mediate this process. In this study, we obtained Jin A and Yamian A, two CMS lines with different cytoplasmic sources.ResultsCompared with maintainer Jin B, Jin A anthers showed advanced tapetal PCD with DNA fragmentation, producing excessive ROS which accumulated around the cell membrane, intercellular space and mitochondrial membrane. The activities of peroxidase (POD) and catalase (CAT) enzymes which can scavenge ROS were significantly decreased. However, Yamian A tapetal PCD was delayed with lower ROS content, and the activities of superoxide dismutase (SOD) and POD were higher than its maintainer. These differences in ROS scavenging enzyme activities may be caused by isoenzyme gene expressions. In addition, we found the excess ROS generated in Jin A mitochondria and ROS overflow from complex III might be the source in parallel with the reduction of ATP content.ConclusionROS accumulation or abrogation were mainly caused by the joint action of ROS generation and scavenging enzyme activities transformation, which led to the abnormal progression of tapetal PCD, affected the development of microspores, and eventually contributed to male sterility. In Jin A, tapetal PCD in advance might be caused by mitochondrial ROS overproduction, accompanied by energy deficiency. The above studies will provide new insights into the cotton CMS and guide the follow-up research ideas.

Salinity effects on the activities of ROS scavenging enzymes in leaves of two sweet potato clones

AbstractSweet potato production, particularly in coastal areas is often prone to salinity. Salt‐tolerant clones will be needed to maintain production, but to date, little is known about salt tolerance traits in sweet potato. Salt stress may result in excessive uptake of unwanted ions into plant tissues leading to the formation of reactive oxygen species (ROS), which in turn may destroy membranes and reduce photosynthesis and growth. Antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POX), catalase (CAT), glutathione reductase (GR) and ascorbate peroxidase (APX) scavenge ROS and early changes in the activities of such enzymes could be used to identify salinity tolerant genotypes. Therefore, cuttings of two contrasting cultivars of sweet potato, BARI SP 8 (tolerant) and BARI SP 4 (sensitive) were greenhouse‐cultivated in nutrient solution for 21 days and then exposed to 100 mmol NaCl for 7 days. Three, five and seven days after salt application the youngest leaves were sampled individually and enzyme activities, potassium (K) and sodium (Na) concentrations, and SPAD (as a proxy for chlorophyll content) were determined. In both varieties leaf growth was not affected by salinity and young leaves grown under salinity had higher SPAD values than older leaves. Na concentration increased over time, particularly in earlier and in older leaves, whereas K was reduced in younger leaves. In general, enzyme activities were strongly affected by leaf age and leaf position. SOD and APX showed varietal but no salinity effects, CAT increased under salinity in both varieties, whereas POX was strongly reduced and GR was strongly increased under salinity in BARI SP 8 with no effect in BARI SP 4. Enzyme activities were not correlated to leaf Na, neither in relation to leaf age, nor leaf number or duration of salt stress in both varieties. However, varietal differences were observed regarding leaf K. Activities of SOD were highly positive and of CAT highly negatively correlated with leaf K under salinity in BARI SP 8 but not in BARI SP 4, whereas activities of GR and POX were strongly positively correlated with leaf K in BARI SP 4 under salinity but not in BARI SP 8. We conclude that potassium may have a strong regulating role on leaf stress levels and therefore on the activities of antioxidant enzymes. Varieties may differ in their tolerance strategy and we have shown that salinity does not generally increase levels of ROS‐scavenging enzymes in sweet potato leaves under salt stress. Confounding factors such as leaf age and leaf position as well as maintaining high leaf level K concentrations need to be considered when evaluating metabolic traits for salinity tolerance traits.

Unraveling the Mechanism of StWRKY6 in Potato (Solanum tuberosum)'s Cadmium Tolerance for Ensuring Food Safety.

The WRKY transcription factor plays a crucial role in plant stress adaptation. Our research has found that WRKY6 in Solanum tuberosum (potatoes) is closely related to cadmium (Cd) tolerance. Therefore, investigating the mechanism of StWRKY6 in plant resistance to Cd toxicity is of great scientific importance for food safety. This research further analyzed the gene structure and functional regions of the nuclear transcription factor WRKY6 in potatoes, discovering that StWRKY6 contains W box, GB/box, ABRE, and other elements that can act as a nuclear transcription regulatory factor to execute multiple functional regulations. The results of the heterologous expression of StWRKY6 in Arabidopsis under Cd stress showed that the overexpression line (StWRKY6-OE) had significantly higher SAPD values and content of reactive oxygen species scavenging enzymes than the wild type, indicating that StWRKY6 plays a crucial role in protecting the photosynthetic system and promoting carbohydrate synthesis. Transcriptome analysis also revealed that the Cd-induced expression of StWRKY6 up-regulated many potential gene targets, including APR2, DFRA, ABCG1, VSP2, ERF013, SAUR64/67, and BBX20, which are involved in Cd chelation (APR2, DFRA), plant defense (VSP2, PDF1.4), toxic substance efflux (ABCG1), light morphology development (BBX20), and auxin signal (SAUR64/67). These genes coordinate the regulation of Cd tolerance in the StWRKY6 overexpression line. In summary, this study identified a potential gene set of the co-expression module of StWRKY6, providing useful evidence for the remediation of Cd-contaminated soil and the genetic breeding of low Cd-accumulating crops, thereby ensuring food safety.

Physiological Variables Influenced by 'Candidatus Liberibacter asiaticus' Infection in Two Citrus Species.

'Candidatus Liberibacter asiaticus' is the bacterium associated with the citrus disease known as huanglongbing (HLB). This study evaluated the influence of 'Ca. L. asiaticus' infection on a number of key plant physiological variables concerning photosynthesis, cell integrity, reactive oxygen species scavengers' activity, and osmoregulation of two different species of citrus-the pomelo Citrus maxima and the mandarin C. reticulata 'Tankan'-relative to their measured 'Ca. L. asiaticus' infection load. Results indicated that all measured physiological variables except soluble sugar were affected by increased 'Ca. L. asiaticus' infection titers, wherein the variety C. maxima proved overall more resistant than C. reticulata. 'Ca. L. asiaticus' infection was linked in both plants to decrease in chlorophyll concentration, cell membrane permeability, and malondialdehyde, as well as increased free proline and starch contents. Chlorophyll fluorescence measurements taken 9 months after grafting the mandarin C. reticulata with 'Ca. L. asiaticus' scions revealed a significant decrease in the photosynthesis variables maximum photochemical quantum yield of photosystem II (PSII), effective photochemical quantum yield of PSII, and coefficient of photochemical fluorescence quenching assuming interconnected PSII antennae, whereas nonphotochemical fluorescence quenching increased significantly; C. maxima plants, on the other hand, did not show significant differences until the 12th month from infection exposure. The variables superoxide dismutase, catalase, peroxidase, and soluble protein initially increased and later decreased. In addition, progression of 'Ca. L. asiaticus' replication in both citrus species was accompanied by rapid changes in three reactive oxygen species scavenging enzymes in C. maxima, while the pattern was different in C. reticulata. We hypothesize that the observed interspecific differences in physiological change are related to their relative resistance against 'Ca. L. asiaticus' infection. These results provide a scaffold for better describing the pathogenesis, selecting the most resistant breeds, or even validating pertaining omics research; ultimately, these detailed observations can facilitate the diagnosis of 'Ca. L. asiaticus' infection.

White Collar 1 Modulates Oxidative Sensitivity and Virulence by Regulating the HOG1 Pathway in Fusarium asiaticum.

Fusarium asiaticum is an epidemiologically important pathogen of cereal crops in east Asia, accounting for both yield losses and mycotoxin contamination problems in food and feed products. FaWC1, a component of the blue-light receptor White Collar complex (WCC), relies on its transcriptional regulatory zinc finger domain rather than the light-oxygen-voltage domain to regulate pathogenicity of F. asiaticum, although the downstream mechanisms remain obscure. In this study, the pathogenicity factors regulated by FaWC1 were analyzed. It was found that loss of FaWC1 resulted in higher sensitivity to reactive oxygen species (ROS) than in the wild type, while exogenous application of the ROS quencher ascorbic acid restored the pathogenicity of the ΔFawc1 strain to the level of the wild type, indicating that the reduced pathogenicity of the ΔFawc1 strain is due to a defect in ROS tolerance. Moreover, the expression levels of the high-osmolarity glycerol (HOG) mitogen-activated protein kinase (MAPK) pathway genes and their downstream genes encoding ROS scavenging enzymes were downregulated in the ΔFawc1 mutant. Upon ROS stimulation, the FaHOG1-green fluorescent protein (GFP)-expressing signal driven by the native promoter was inducible in the wild type but negligible in the ΔFawc1 strain. Overexpressing Fahog1 in the ΔFawc1 strain could recover the ROS tolerance and pathogenicity of the ΔFawc1 mutant, but it remained defective in light responsiveness. In summary, this study dissected the roles of the blue-light receptor component FaWC1 in regulating expression levels of the intracellular HOG-MAPK signaling pathway to affect ROS sensitivity and pathogenicity in F. asiaticum. IMPORTANCE The well-conserved fungal blue-light receptor White Collar complex (WCC) is known to regulate virulence of several pathogenic species for either plant or human hosts, but how WCC determines fungal pathogenicity remains largely unknown. The WCC component FaWC1 in the cereal pathogen Fusarium asiaticum was previously found to be required for full virulence. The present study dissected the roles of FaWC1 in regulating the intracellular HOG MAPK signaling pathway to affect ROS sensitivity and pathogenicity in F. asiaticum. This work thus extends knowledge of the association between fungal light receptors and the intracellular stress signaling pathway to regulate oxidative stress tolerance and pathogenicity in an epidemiologically important fungal pathogen of cereal crops.

Melatonin-induced myeloblastosis viral oncogene homologs alleviate fresh-cut lotus root browning during storage by attenuating flavonoid biosynthesis and reactive oxygen species.

Lotus roots (Nelumbo nucifera Gaertn.) are rich in nutrients and have ornamental and food value. However, browning has caused huge economic losses and security risks during the storage and harvesting of fresh-cut lotus. This study investigated the role of melatonin in inhibiting lotus browning, and illustrates its molecular mechanism. The application of melatonin effectively retarded the process of lotus browning, enhanced reactive oxygen species (ROS) scavenging enzyme activity, and inhibited the activity of polyphenol oxidase (PPO), and peroxidase (POD). Melatonin reduced flavonoid content, and decreased enzymatic activity in flavonoid biosynthesis. Transcriptome Sequencing (RNA-seq) was used to screen the genes regulated by exogenous melatonin when defending against fresh-cut lotus browning. Gene co-expression analysis (GCN) indicated that the transcription factors MYB5, MYB6, and MYB308, activated by melatonin, were negatively related to the expression of PPO and the genes related to flavonoid and phenylpropanoid biosynthesis. These myeloblastosis viral oncogene homologs (MYBs) were positively related to the expression of genes encoding the enzymes in glutathione metabolism. Melatonin retarded lotus browning by transcriptional suppression of key genes associated with flavonoid and phenylpropanoid biosynthesis through the stimulation of MYB5, MYB6, and MYB308. © 2023 Society of Chemical Industry.

A novel module regulating ROS in NLR-mediated immunity.

The regulatory mechanisms of apoplastic reactive oxygen species (ROS) production during pattern-triggered immunity (PTI) are well known. However, how ROS levels are regulated during effector-triggered immunity (ETI) remains largely unknown. Recently, Zhang et al. discovered that MAPK-Alfin-like 7 module enhances nucleotide-binding, leucine-rich repeat receptor (NLR)-mediated immunity by negatively regulating genes encoding ROS scavenging enzymes, deepening our understanding of ROS control during ETI in plants.