Rate Of Superoxide Production Research Articles

Electron-beam generated X-ray irradiation could retard the senescence of postharvest Hericium erinaceus via regulating reactive oxygen metabolism

Fresh Hericium erinaceus, a type of white mushroom, is prone to browning and microbial invasion after harvest due to high rate of metabolism. This study aimed to clarify how electron-beam generated X-ray (EBGX) irradiation (0, 1.0, 1.5, and 2.0 kGy) affects the senescence of postharvest fresh H. erinaceus during storage at 15 °C for 9 d. The effects of EBGX irradiation on biological properties of H. erinaceus were evaluated. Results indicated that EBGX irradiation reduced the incidence of browning in H. erinaceus, increased firmness, inhibited the respiration rate, lowered malondialdehyde (MDA) content and electrolyte leakage relative to untreated samples. Moreover, the natural microflora presents on H. erinaceus were prominently inhibited (P < 0.05) in 1.0 kGy compare to control group after storage, such as total bacterial count (35.6%), molds and yeasts (31.4%), and Pseudomonas (33.2%). Meanwhile, exposure to 1.0 kGy of irradiation treatment exhibited a minimum hydrogen peroxide content (51.2 nmol kg−1) and superoxide anion production rate (1.88 μmol kg−1 min−1) at 9 d, enhanced the activities of antioxidative enzymes (superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase), as well as declined oxidative stress by regulating the expression of related genes (HeSOD1, HeSOD2, HeCAT1, HeCAT2, HePOX1, and HePOX2).TUNEL analysis also observed that H. erinaceus treated with 1.0 kGy remarkably retained a lower nuclear DNA fragmentation. This finding suggests that the dose of 1.0 kGy EBGX irradiation could postpone the senescence process of fresh H. erinaceus and has the potential to be a promising physical preservation technology for fresh H. erinaceus.

Integrative leaf anatomy structure, physiology, and metabolome analyses revealed the response to drought stress in sainfoin at the seedling stage.

Sainfoin (Onobrychis viciaefolia) is a vital legume forage, and drought is the primary element impeding sainfoin growth. The anatomical structure, physiological indexes, and metabolites of the leaves of sainfoin seedlings with a drought-resistant line of P1 (DRL) and a drought-sensitive material of 2049 (DSM) were analyzed under drought (-1.0 MPa) with polyethylene glycol-6000 (PEG-6000). The leaf anatomy was studied by the paraffin section method. The related physiological indexes were measured by the hydroxylamine oxidation method, titanium sulfate colorimetric method, thiobarbituric acid method, acidic ninhydrin colorimetric method, and Coomassie brilliant blue method. The metabolomics analysis was composed of liquid chromatography tandem high-resolution mass spectrometry (LC-MS/MS). The results revealed that the thickness of the epidermis, palisade tissue, and sponge tissue of DRL were significantly greater than those of DSM. The leaves of DRL exhibited lower levels of superoxide anion (O2 •-) production rate, hydrogen peroxide (H2O2) content, and malondialdehyde (MDA) content compared with DSM, while proline (Pro) content and soluble protein (SP) content were significantly higher than those of DSM. A total of 391 differential metabolites were identified in two samples. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment showed that the primary differential metabolites were concentrated into the tyrosine metabolism; isoquinoline alkaloid biosynthesis; ubiquinone and other terpenoid quinone biosynthesis; neomycin, kanamycin, and gentamicin biosynthesis; and anthocyanin biosynthesis metabolic pathways. Compared with DSM, DRL had more complete anatomical structure, lower active oxygen content, and higher antioxidant level. The results improved our insights into the drought-resistant mechanisms in sainfoin.

Plant growth regulators mitigate oxidative damage to rice seedling roots by NaCl stress.

The aim of this experiment was to investigate the effects of exogenous sprays of 5-aminolevulinic acid (5-ALA) and 2-Diethylaminoethyl hexanoate (DTA-6) on the growth and salt tolerance of rice (Oryza sativa L.) seedlings. This study was conducted in a solar greenhouse at Guangdong Ocean University, where 'Huanghuazhan' was selected as the test material, and 40 mg/L 5-ALA and 30 mg/L DTA-6 were applied as foliar sprays at the three-leaf-one-heart stage of rice, followed by treatment with 0.3% NaCl (W/W) 24 h later. A total of six treatments were set up as follows: (1) CK: control, (2) A: 40 mg⋅ L-1 5-ALA, (3) D: 30 mg⋅ L-1 DTA-6, (4) S: 0.3% NaCl, (5) AS: 40 mg⋅ L-1 5-ALA + 0.3% NaCl, and (6) DS: 30 mg⋅ L-1 DTA-6+0.3% NaCl. Samples were taken at 1, 4, 7, 10, and 13 d after NaCl treatment to determine the morphology and physiological and biochemical indices of rice roots. The results showed that NaCl stress significantly inhibited rice growth; disrupted the antioxidant system; increased the rates of malondialdehyde, hydrogen peroxide, and superoxide anion production; and affected the content of related hormones. Malondialdehyde content, hydrogen peroxide content, and superoxide anion production rate significantly increased from 12.57% to 21.82%, 18.12% to 63.10%, and 7.17% to 56.20%, respectively, in the S treatment group compared to the CK group. Under salt stress, foliar sprays of both 5-ALA and DTA-6 increased antioxidant enzyme activities and osmoregulatory substance content; expanded non-enzymatic antioxidant AsA and GSH content; reduced reactive oxygen species (ROS) accumulation; lowered malondialdehyde content; increased endogenous hormones GA3, JA, IAA, SA, and ZR content; and lowered ABA content in the rice root system. The MDA, H2O2, and O2- contents were reduced from 35.64% to 56.92%, 22.30% to 53.47%, and 7.06% to 20.01%, respectively, in the AS treatment group compared with the S treatment group. In the DS treatment group, the MDA, H2O2, and O2- contents were reduced from 24.60% to 51.09%, 12.14% to 59.05%, and 12.70% to 45.20%. In summary, NaCl stress exerted an inhibitory effect on the rice root system, both foliar sprays of 5-ALA and DTA-6 alleviated damage from NaCl stress on the rice root system, and the effect of 5-ALA was better than that of DTA-6.

Hydrogen Sulfide Alleviates Oxidative Damage under Chilling Stress through Mitogen-Activated Protein Kinase in Tomato.

Tomato is the vegetable with the largest greenhouse area in China, and low temperature is one of the main factors affecting tomato growth, yield, and quality. Hydrogen sulfide (H2S) plays an important role in regulating plant chilling tolerance, but its downstream cascade reaction and mechanism remain unclear. Mitogen-activated protein kinases (MAPK/MPKs) are closely related to a variety of signaling substances in stress signal transmission. However, whether H2S is related to the MPK cascade pathway in response to low-temperature stress is rarely reported. In this study, NaHS treatment significantly decreased the electrolyte leakage (EL), superoxide anion (O2-) production rate, and hydrogen peroxide (H2O2) content of seedlings at low temperatures. In addition, the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were obviously increased; and the photochemical efficiency of PSII (Fv/Fm) was enhanced with treatment with NaHS, indicating that NaHS improved the seedlings' cold tolerance by alleviating the degree of membrane lipid peroxidation and oxidative damage. However, H2S scavenger hypotaurine (HT) treatment showed the opposite effect. We found that H2S content, L-cysteine desulfhydrase (LCD) activity, and mRNA expression were increased by chilling stress but reduced by MPK inhibitor PD98059; PD98059 reversed the alleviating effect of H2S via increasing the EL and H2O2 contents. The expression levels of MPK1-MPK7 at low temperatures showed that SlMPK4 was significantly induced by exogenous NaHS and showed a trend of first increasing and then decreasing, while the expression level of SlMPK4 in HT-treated seedlings was lower than that of the control. After SlMPK4 was silenced by virus-induced gene silencing, the H2S-induced upregulation of C-repeat-Binding Factor (CBF1), inducer of CBF expression 1 (ICE1), respiratory burst oxidase homologs (RBOH1, RBOH2) at low temperatures disappeared, and tomato cold tolerance decreased. In conclusion, H2S improves the cold tolerance of tomato plants by increasing the activity of antioxidant enzymes and reducing reactive oxygen species (ROS) accumulation and membrane lipid peroxidation. MPK4 may act as a downstream signaling molecule in this process.

Boron Reduced Copper Excess-Induced Oxidative Damage in Citrus sinensis by Modulating Reactive Oxygen Species and Methylglyoxal Formation and Their Detoxification Systems.

Citrus is mainly cultivated in acid soil with low boron (B) and high copper (Cu). In this study, Citrus sinensis seedlings were submitted to 0.5 (control) or 350 μM Cu (Cu excess or Cu exposure) and 2.5, 10, or 25 μM B for 24 weeks. Thereafter, H2O2 production rate (HPR), superoxide production rate (SAPR), malondialdehyde, methylglyoxal, and reactive oxygen species (ROS) and methylglyoxal detoxification systems were measured in leaves and roots in order to test the hypothesis that B addition mitigated Cu excess-induced oxidative damage in leaves and roots by reducing the Cu excess-induced formation and accumulation of ROS and MG and by counteracting the impairments of Cu excess on ROS and methylglyoxal detoxification systems. Cu and B treatments displayed an interactive influence on ROS and methylglyoxal formation and their detoxification systems. Cu excess increased the HPR, SAPR, methylglyoxal level, and malondialdehyde level by 10.9% (54.3%), 38.9% (31.4%), 50.3% (24.9%), and 312.4% (585.4%), respectively, in leaves (roots) of 2.5 μM B-treated seedlings, while it only increased the malondialdehyde level by 48.5% (97.8%) in leaves (roots) of 25 μM B-treated seedlings. Additionally, B addition counteracted the impairments of Cu excess on antioxidant enzymes, ascorbate-glutathione cycle, sulfur metabolism-related enzymes, sulfur-containing compounds, and methylglyoxal detoxification system, thereby protecting the leaves and roots of Cu-exposed seedlings against oxidative damage via the coordinated actions of ROS and methylglyoxal removal systems. Our findings corroborated the hypothesis that B addition alleviated Cu excess-induced oxidative damage in leaves and roots by decreasing the Cu excess-induced formation and accumulation of ROS and MG and by lessening the impairments of Cu excess on their detoxification systems. Further analysis indicated that the pathways involved in the B-induced amelioration of oxidative stress caused by Cu excess differed between leaves and roots.

Defense Responses of Different Rice Varieties Affect Growth Performance and Food Utilization of Cnaphalocrocis medinalis Larvae

Rice leaf folder, Cnaphalocrocis medinalis (Guenée), is one of the most serious pests on rice. At present, chemical control is the main method for controlling this pest. However, the indiscriminate use of chemical insecticides has non-target effects and may cause environmental pollution. Besides, leaf curling behavior by C. medinalis may indirectly reduce the efficacy of chemical spray. Therefore, it is crucial to cultivate efficient rice varieties resistant to this pest. Previous studies have found that three different rice varieties, Zhongzao39 (ZZ39), Xiushui134 (XS134), and Yongyou1540 (YY1540), had varying degrees of infestation by C. medinalis. However, it is currently unclear whether the reason for this difference is related to the difference in defense ability of the three rice varieties against the infestation of C. medinalis. To explore this issue, the current study investigated the effects of three rice varieties on the growth performance and food utilization capability of the 4th instar C. medinalis. Further, it elucidated the differences in defense responses among different rice varieties based on the differences in leaf physiological and biochemical indicators and their impact on population occurrence. The results showed that the larval survival rate was the lowest, and the development period was significantly prolonged after feeding on YY1540. This was not related to the differences in leaf wax, pigments, and nutritional components among the three rice varieties nor to the feeding preferences of the larvae. The rate of superoxide anion production, hydrogen peroxide content, and the activity of three protective enzymes were negatively correlated with larval survival rate, and they all showed the highest in YY1540 leaves. Compared to other tested varieties, although the larvae feeding on YY1540 had higher conversion efficiency of ingested food and lower relative consumption rate, their relative growth was faster, indicating stronger food utilization capability. However, they had a lower accumulation of protein. This suggests that different rice varieties had different levels of oxidative stress after infestation by C. medinalis. The defense response of YY1540 was more intense, which was not conducive to the development of the larvae population. These results will provide new insights into the interaction mechanism between different rice varieties and C. medinalis and provide a theoretical basis for cultivating rice varieties resistant to this pest.

Foliar spraying of exogenous uniconazole (S3307) at the flowering stage as an effective method to resist low-temperature stress on mung bean [Vigna radiata (L.) Wilczek

Low temperature is one of the major constraints on agricultural productivity worldwide and is likely to further increase. Several adaptations and mitigation strategies are required to cope with low-temperature stress. Uniconazole (S3307) could play a significant role in the alleviation of abiotic stress in plants. In this study, the effects of S3307 on the reactive oxygen species (ROS) and antioxidant metabolism were studied in the leaves of mung bean [Vigna radiata (L.) Wilczek]. The experimental results showed that the low-temperature induced accumulation of superoxide anion (O2-) production rate, and malonaldehyde (MDA) contents. Increased proline content and enzymatic antioxidants, including superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), were found to alleviate oxidative damage under low temperatures. While, S3307 could reduce O2- production rate and MDA contents and increase the activities of SOD, POD, and CAT, slowed the decrease in ascorbic acid (AsA), dehydroascorbic acid (DHA), glutathione (GSH), and oxidized glutathione (GSSG), and promoted increase in soluble sugars (SS), soluble proteins (SP), and proline (Pro) content under low-temperature. At the same time, low temperature leads to lower 100 grain weight and number of grains per plant, which eventually causes yield reduction decreased. Foliar spraying of S3307 could alleviate the yield loss caused by low temperature, and the increase of S3307 treatment was 5.1%–12.5% and 6.3%–32.9% for the two varieties, respectively, compared with CK. In summary, exogenous S3307 pretreatment enhances plant tolerance to low-temperature by improving the antioxidant enzyme activities, increased non-enzymatic antioxidants content, and decreased O2- production rate and MDA contents and inducing alterations in endogenous S3307, and reduce the decrease in mung bean yield.

Markers of oxidative stress during post-COVID-19 fatigue: a hypothesis-generating, exploratory pilot study on hospital employees.

Post-COVID-19 fatigue is common after recovery from COVID-19. Excess formation of reactive oxygen species (ROS) leading to oxidative stress-related mitochondrial dysfunction is referred to as a cause of these chronic fatigue-like symptoms. The present observational pilot study aimed to investigate a possible relationship between the course of ROS formation, subsequent oxidative stress, and post-COVID-19 fatigue. A total of 21 post-COVID-19 employees of the General Hospital Nuremberg suffering from fatigue-like symptoms were studied during their first consultation (T1: on average 3 months after recovery from COVID-19), which comprised an educational talk on post-COVID-19 symptomatology and individualized outpatient strategies to resume normal activity, and 8 weeks thereafter (T2). Fatigue severity was quantified using the Chalder Fatigue Scale together with a health survey (Patient Health Questionnaire) and self-report on wellbeing (12-Item Short-Form Health Survey). We measured whole blood superoxide anion () production rate (electron spin resonance, as a surrogate for ROS production) and oxidative stress-induced DNA strand breaks (single cell gel electrophoresis: "tail moment" in the "comet assay"). Data are presented as mean ± SD or median (interquartile range) depending on the data distribution. Differences between T1 and T2 were tested using a paired Wilcoxon rank sign or t-test. Fatigue intensity decreased from 24 ± 5 at T1 to 18 ± 8 at T2 (p < 0.05), which coincided with reduced formation (from 239 ± 55 to 195 ± 59 nmol/s; p < 0.05) and attenuated DNA damage [tail moment from 0.67 (0.36-1.28) to 0.32 (0.23-0.71); p = 0.05]. Our pilot study shows that post-COVID-19 fatigue coincides with (i) enhanced formation and oxidative stress, which are (ii) reduced with attenuation of fatigue symptoms.

Nitrogen-deficient leaves and roots can keep high abilities to scavenge reactive oxygen species and methylglyoxal, and protect them against oxidative damage in Citrus sinensis seedlings

Nitrogen-deficiency (ND) usually occurs in some citrus orchard soils in China. The roles of reactive oxygen species (ROS) and methylglyoxal (MG) generation and their detoxification systems in ND tolerance of horticultural woody plants still need to be revealed. For the first time, we examined the effects of ND on ROS and MG generation and their detoxification systems in leaves and roots of Citrus sinensis seedlings. The objectives are to test the hypotheses that N-deficient leaves and roots can keep high abilities to scavenge ROS and MG, thereby protecting them from oxidative damage, and that ND-induced alterations of ROS and MG formation and their detoxification systems in leaves and roots are different. ND augmented superoxide anion production rate and MG concentrations, but it decreased malondialdehyde (MDA) concentrations and electrolyte leakage in leaves and roots. ND increased the activities of most enzymes involved in ROS (ascorbate-glutathione cycle-related enzymes, antioxidant enzymes, and sulfur metabolism-related enzymes) and MG (glyoxalases) detoxification expressed on a protein basis with a few exceptions, and the concentrations of ascorbate, phytochelatins, and total non-protein thiols in leaves and roots. These results suggested that nitrogen-deficient leaves and roots could keep high abilities to detoxify ROS and MG, and protect them from oxidative damage. Generally viewed, ND affected the production and removal of ROS and MG more in roots than in leaves.

Changes in Activity of Antioxidant Systems of &lt;i&gt;Escherichia coli&lt;/i&gt; under Phosphate Starvation

Changes in the activity of antioxidant systems in Escherichia coli during phosphate starvation were studied. It was shown that starvation was accompanied by a decrease in the intensity of respiration, an increase in the rate of superoxide production, and a decrease in the level of ATP. Simultaneously, there was a decrease in H2O2 in the medium and a significant increase in the expression of the katG and katE genes encoding the HPI and HPII catalases, respectively. At the same time, there was no drop in the membrane potential, which may indicate the retention of normal membrane activity in starving cells. It has been shown for the first time that the transition of E. coli to phosphate starvation is accompanied by significant changes in the status of glutathione. The most important of them are associated with a decrease in the level of glutathione reductive form (GSH) in the medium (GSHout) and with a simultaneous increase in its content in the cytoplasm (GSHin), as well as a shift in the GSHin to oxidized glutathione form (GSSGin) ratio towards reductive values, and GSHout/GSSGout towards oxidative values. Among the mutants used in the work, the double mutant gor trxB, deficient in the synthesis of glutathione reductase and thioredoxin reductase, showed the most pronounced distinctive features. Compared to the parental strain, this mutant showed a multiple higher expression of katG::lacZ, the highest level of oxidized intra- and extracellular glutathione, and, accordingly, the lowest GSH/GSSG ratio in both compartments. In general, the data obtained indicate that during phosphate starvation the interaction of the glutathione redox-system and regulons that control protection against reactive oxygen species creates conditions that allow maintaining the concentration of ROS below the toxic level. As a result, phosphate-starved E. coli cells can maintain a high viability for a long time that allows them quickly to resume growth after the addition of phosphate.

Assessment of Bacillus subtilis fermented Caulerpa microphysa byproduct as feed additive on the growth performance, immune regulation and disease resistance of white shrimp (Litopenaeus vannamei)

In this study, the immunomodulatory and antioxidant activity of fermented Caulerpa microphysa byproduct (FCMB) by Bacillus subtilis was evaluated, and its potential as a feed additive for white shrimp (Litopenaeus vannamei) was explored. In vitro experiments showed that the FCMB supernatant contained polysaccharides, polyphenols and flavonoids, and exhibited antioxidant properties as assessed by various antioxidant assays. Additionally, the FCMB supernatant was found to increase the production rate of reactive oxygen species and the activity of phenoloxidase in hemocytes in vitro. Furthermore, the results of the in vivo feeding trial showed that dietary 5 g kg−1 FCMB significantly improved the weight gain and specific growth rate of white shrimp after 56 days of feeding. Although there were no significant differences in total hemocyte count, phagocytosis, superoxide anion production rate, and phenoloxidase activity among the experimental groups, upregulation of immune-related genes was observed, particularly in the hepatopancreas and hemocytes of shrimps fed with 5 g or 50 g FCMB per kg feed, respectively. In the pathogen challenge assay, white shrimp fed with 5 % FCMB exhibited a higher survival rate compared to the control group following Vibrio parahaemolyticus challenge. Therefore, it is concluded that the fermented byproduct of C. microphysa, FCMB, holds potential as a feed additive for enhancing the growth performance and disease resistance against V. parahaemolyticus in white shrimp.

Drought resistance index screening and evaluation of lettuce under water deficit conditions on the basis of morphological and physiological differences.

Water is one of the important factors affecting the yield of leafy vegetables. Lettuce, as a widely planted vegetable, requires frequent irrigation due to its shallow taproot and high leaf evaporation rate. Therefore, screening drought-resistant genotypes is of great significance for lettuce production. In the present study, significant variations were observed among 13 morphological and physiological traits of 42 lettuce genotypes under normal irrigation and water-deficient conditions. Frequency analysis showed that soluble protein (SP) was evenly distributed across six intervals. Principal component analysis (PCA) was conducted to transform the 13 indexes into four independent comprehensive indicators with a cumulative contribution ratio of 94.83%. The stepwise regression analysis showed that root surface area (RSA), root volume (RV), belowground dry weight (BDW), soluble sugar (SS), SP, and leaf relative water content (RWC) could be used to evaluate and predict the drought resistance of lettuce genotypes. Furthermore, the drought resistance ranks of the genotypes were similar according to the drought resistance comprehensive evaluation value (D value), comprehensive drought resistance coefficient (CDC), and weight drought resistance coefficient (WDC). The cluster analysis enabled the division of the 42 genotypes into five drought resistance groups; among them, variety Yidali151 was divided into group I as a strongly drought-resistant variety, group II included 6 drought-resistant genotypes, group III included 16 moderately drought-resistant genotypes, group IV included 12 drought-sensitive genotypes, and group V included 7 highly drought-sensitive genotypes. Moreover, a representative lettuce variety was selected from each of the five groups to verify its water resistance ability under water deficit conditions. In the drought-resistant variety, it was observed that stomatal density, superoxide anion (O2.-wfi2) production rate, and malondialdehyde (MDA) content exhibited a low increase rate, while catalase (CAT), superoxide dismutase (SOD), and that peroxidase (POD) activity exhibited a higher increase than in the drought-sensitive variety. In summary, the identified genotypes are important because their drought-resistant traits can be used in future drought-resistant lettuce breeding programs and water-efficient cultivation.

Photosynthetic and Physiological Responses to Combined Drought and Low–Temperature Stress in Poa annua Seedlings from Different Provenances

Combined drought and low–temperature stress is a crucial factor affecting turfgrass establishment and limiting the sustainability of the turfgrass industry in drought– and cold–prone regions. In this context, we evaluated the effects of regular watering (the soil water content was 80% of the maximum water–holding capacity of the field) at room temperature (25 °C) and combined drought (the soil water content was 30% of the maximum water–holding capacity of the field) and low–temperature (0 °C) stress on the morphology, photosynthesis, and physiology of wild Poa annua seedlings from different provenances (‘PA’, ‘WY’, ‘NX’ and ‘YC’). Results indicated that the combined drought and low–temperature stress changed the morphological and growth indicators of seedlings in four provenances to different extents. Moreover, combined drought and low–temperature stress reduced the net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), water use efficiency (WUE), and chlorophyll content in seedlings from four provenances. However, intertemporal CO2 concentration (Ci), relative electrical conductivity (REC), the contents of malondialdehyde (MDA), proline (Pro), soluble sugars (SS), the superoxide anion (O2•−) production rate, the contents of hydrogen peroxide (H2O2) and hydroxyl radical (·OH), the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) were all increased. The increase in ‘PA’ was much greater than that in ‘NX.’ The comprehensive evaluation results showed that the order of combined drought and low–temperature resistance of seedlings from the four provenances was ‘PA’ &gt; ‘YC’ &gt; ‘WY’ &gt; ‘NX’, which corresponded to the order of the morphological damage symptoms. In conclusion, ‘PA’ may maintain stronger combined drought and low–temperature resistance by improving the cellular water absorption and retention capacity, enhancing the function of the antioxidant defense system, and maintaining the integrity of the cell membrane, which is a crucial germplasm resource for breeding combined drought and low–temperature resistance in Poa annua.

Full-Length Transcriptome Sequencing Reveals the Molecular Mechanism of Metasequoia glyptostroboides Seed Responding to Aging.

Metasequoia glyptostroboides, Hu and W. C. Cheng, as the only surviving relict species of the Taxodiaceae Metasequoia genus, is a critically endangered and protected species in China. There is a risk of extinction due to the low vigor of M. glyptostroboides seeds, and the physiological mechanism of seed aging in M. glyptostroboides is not yet clear. In order to investigate the physiological and molecular mechanisms underlying the aging process of M. glyptostroboides seeds, we analyzed the antioxidant system and transcriptome at 0, 2, 4, 6, and 8 days after artificial accelerated aging treatment at 40 °C and 100% relative humidity. It was found that the germination percentage of fresh dried M. glyptostroboides seeds was 54 ± 5.29%, and significantly declined to 9.33 ± 1.88% after 6 days of aging, and then gradually decreased until the seed died on day 8. Superoxide dismutase (SOD) activity, ascorbic acid (AsA), glutathione (GSH) content and superoxide anion (O2·-) content and production rate significantly decreased, while malondialdehyde (MDA) and hydrogen peroxide (H2O2) content and glutathione peroxidase (GPX) and catalase (CAT) activity gradually increased during the aging process. A total of 42,189 unigenes were identified in the whole transcriptome, and 40,446 (95.86%) unigenes were annotated in at least one protein database. A total of 15,376 differentially expressed genes (DEGs) were obtained; KEGG enrichment analysis results revealed that seed aging may be mainly involved in the protein-processing pathways in endoplasmic reticulum, oxidative phosphorylation, and ascorbate and aldarate metabolism. Weighted gene co-expression network analysis (WGCNA) revealed that the dark magenta, orange, and medium purple modules were highly correlated with physiological indicators such as SOD, CAT, and GSH and further identified 40 hub genes such as Rboh, ACO, HSF, and CML as playing important roles in the antioxidant network of M. glyptostroboides seeds. These findings provide a broader perspective for studying the regulatory mechanism of seed aging and a large number of potential target genes for the breeding of other endangered gymnosperms.

Effect of gibberellic acid on photosynthesis and oxidative stress response in maize under weak light conditions.

Gibberellin (GA) is an important endogenous hormone involved in plant responses to abiotic stresses. Experiments were conducted at the Research and Education Center of Agronomy, Shenyang Agricultural University (Shenyang, China) in 2021.We used a pair of near-isogenic inbred maize lines comprising, SN98A (light-sensitive inbred line) and SN98B (light-insensitive inbred line) to study the effects of exogenous gibberellin A3 (GA3) application on different light-sensitive inbred lines under weak light conditions. The concentration of GA3 was selected as 20, 40 and 60 mg L-1. After shade treatment, the photosynthetic physiological indexes of SN98A were always lower than SN98B, and the net photosynthetic rate of SN98A was 10.12% lower than SN98B on the 20th day after shade treatment. GA3 treatments significantly reduced the barren stalk ratios in SN98A and improved its seed setting rates by increasing the net photosynthetic rate (Pn), transpiration rate (Tr), stomatal conductance (Gs), photosynthetic pigment contents, photochemical efficiency of photosystem II (PS II) (Fv/Fm), photochemical quenching coefficient (qP), effective quantum yield of PSII photochemistry (ΦPSII), and antioxidant enzyme activities, where the most effective treatment was 60 mg L-1GA3. Compared with CK group, the seed setting rate increased by 33.87%. GA3 treatment also regulated the metabolism of reactive oxygen species (ROS) and reduced the superoxide anion ( ) production rate, H2O2 content, and malondialdehyde content. The superoxide anion ( ) production rate, H2O2 content and malondialdehyde content of SN98A sprayed with 60 mg L-1 GA3 decreased by 17.32%,10.44% and 50.33% compared with CK group, respectively. Compared with the control, GA3 treatment significantly (P < 0.05) increased the expression levels of APX and GR in SN98A, and APX, Fe-SOD, and GR in SN98B. Weak light stress decreased the expression of GA20ox2, which was related to gibberellin synthesis, and the endogenous gibberellin synthesis of SN98A. Weak light stress accelerated leaf senescence, and exogenous GA3 application inhibited the ROS levels in the leaves and maintained normal physiological functions in the leaves. These results indicate that exogenous GA3 enhances the adaptability of plants to low light stress by regulating photosynthesis, ROS metabolism and protection mechanisms, as well as the expression of key genes, which may be an economical and environmentally friendly method to solve the low light stress problem in maize production.

Identification of the BcLEA Gene Family and Functional Analysis of the BcLEA73 Gene in Wucai (Brassica campestris L.).

Late embryogenesis abundant (LEA) proteins are important developmental proteins in the response of plants to abiotic stress. In our previous study, BcLEA73 was differentially expressed under low-temperature stress. Herein, we combined bioinformatics analysis, subcellular localization, expression assays, and stress experiments (including salt, drought, and osmotic stress) to identify and analyze the BcLEA gene family. Gene cloning and functional analysis of BcLEA73 were performed in tobacco and Arabidopsis. Based on the sequence homology and the available conservative motif, 82 BrLEA gene family members were identified and were divided into eight subfamilies in the genome-wide database of Chinese cabbage. The analysis showed that the BrLEA73 gene was located on chromosome A09 and belonged to the LEA_6 subfamily. Quantitative real-time PCR analysis indicated that the BcLEA genes were differentially expressed to varying degrees in the roots, stems, leaves, and petioles of Wucai. The overexpressed BcLEA73 transgenic plants exhibited no significant differences in root length and seed germination rates compared to the wild-type (WT) plants under control conditions. Under salt and osmotic stress treatment, the root length and seed germination rates of the BcLEA73-OE strain were significantly greater than those of WT plants. Under salt stress, the total antioxidant capacity (T-AOC) of the BcLEA73-OE lines increased significantly, and the relative conductivity, (REL), hydrogen peroxide (H2O2) content, and superoxide anion (O2-) production rate decreased significantly. Under drought treatment, the survival rate of the BcLEA73-OE lines was significantly higher than that of WT plants. These results showed that the BcLEA73 gene of Wucai functions in enhancing the tolerance of plants to salt, drought, and osmotic stress. This study provides a theoretical basis to explore the relevant functions of the BcLEA gene family members of Wucai.

TrPLD1 and TrPLD2 modulate reactive oxygen species production and pathogenicity in Trichothecium roseum infected apple fruit

Phospholipase D (PLD) plays an important role during pathogen-host interaction, which not only hydrolyzes the phosphodiester bond in the phospholipid structure of the host cell membrane, resulting in the cell membrane structure destruction of the host, but also produces phosphatidic acid that can interact with NADPH oxidase (NOX) on the cell plasma membrane, resulting in reactive oxygen species (ROS) accumulation and improving the pathogenicity of the pathogen. However, the ROS production of filamentous fungi by PLD modulation and its pathogenicity on postharvest fruit and vegetables have not been documented. In this study, we constructed ΔTrPLD1 and ΔTrPLD2 mutants and complement strains of Trichothecium roseum by homologous recombination, and analyzed ROS production and its pathogenicity on apple fruit. The results showed that the deletion of TrPLD1 and TrPLD2 increased the sensitivity to oxidative stress and reduced ROS production of T. roseum mycelia. During ΔTrPLD1 mutant of T. roseum infecting apple fruit, the gene expression and activity of NOX decreased, which lead to the reduction of superoxide anion production rate. The gene expressions and activities of superoxide dismutase and peroxidase were up-regulated, the expression and activity of catalase were down-regulated, and the activation of ascorbic acid-glutathione cycle, ultimately enhanced the production of H2O2 in apple tissue, H2O2 acted as signal molecule to trigger disease resistance of fruit, thereby reducing the pathogenicity on apple fruit. The ΔTrPLD2 mutant had no significant effect on ROS production during pathogen infecting apple fruit. In summary, ΔTrPLD1 and ΔTrPLD2 mutants were highly sensitive to oxidative stress, and TrPLD1 played more important role in T. roseum infected apple fruit by regulating ROS production.

The evolution of the human mitochondrial bc1 complex- adaptation for reduced rate of superoxide production?

The mitochondrial bc1 complex is a major source of mitochondrial superoxide. While bc1-generated superoxide plays a beneficial signaling role, excess production of superoxide lead to aging and degenerative diseases. The catalytic core of bc1 comprises three peptides -cytochrome b, Fe-S protein, and cytochrome c1. All three core peptides exhibit accelerated evolution in anthropoid primates. It has been suggested that the evolution of cytochrome b in anthropoids was driven by a pressure to reduce the production of superoxide. In humans, the bc1 core peptides exhibit anthropoid-specific substitutions that are clustered near functionally critical sites that may affect the production of superoxide. Here we compare the high-resolution structures of bovine, mouse, sheep and human bc1 to identify structural changes that are associated with human-specific substitutions. Several cytochrome b substitutions in humans alter its interactions with other subunits. Most significantly, there is a cluster of seven substitutions, in cytochrome b, the Fe-S protein, and cytochrome c1 that affect the interactions between these proteins at the tether arm of the Fe-S protein and may alter the rate of ubiquinone oxidation and the rate of superoxide production. Another cluster of substitutions near heme bH and the ubiquinone reduction site, Qi, may affect the rate of ubiquinone reduction and thus alter the rate of superoxide production. These results are compatible with the hypothesis that cytochrome b in humans (and other anthropoid primates) evolve to reduce the rate of production of superoxide thus enabling the exceptional longevity and exceptional cognitive ability of humans.

Mitigation of exogenous melatonin on photoinhibition of tomato seedlings under chilling stress.

To understand the mitigation effects of melatonin on the chilling-induced photoinhibition in tomato, four groups of seedlings were labelled: NW (normal temperature + water), NM (normal temperature + melatonin), CW (chilling + water) and CM (chilling+ melatonin). We measured chlorophyll fluorescence, key photosynthetic parameters and the cycle efficiency for chloroplast ascorbic acid-glutathione (AsA-GSH). The results showed that, compared with the NW control, photosynthesis rate in CW was decreased by 50.3%-72.6%, chloroplast malondialdehyde content was decreased by 17.5%-132.7%, superoxide anion production was increased by 86.5%-235.9%, and H2O2 was increased by 96.6%-208.4%. These trends were significantly alleviated by exogenous melatonin, with photosynthetic rates in CM being increased by 22.7%-24.7% compared with in CW, malondialdehyde content being decreased by 16.6%-29.0%, the rate of superoxide anion production being decreased by 14.9%-22.7%, H2O2 content being decreased by 10.7%-27.1%. Compared with CW, the quantum yield of photochemical energy in PS Ⅱ was increased by 15.8% in CM, the quantum yield of regulated non-photochemical energy loss was increased by 7.2%, the quantum yield of non-regulated non-photochemical energy loss was decreased by 24.7%, and the activities of key metabolic enzymes in the AsA-GSH cycle were increased to different degrees. We concluded that exogenous melatonin application could alleviate photoinhibition in tomato seedlings under chilling by balancing the partitioning of absorption energy in PS Ⅱ and by enhancing the ROS scavenging efficiency of the AsA-GSH cycle in the chloroplast.

Fucoidan treatment alleviates chilling injury in cucumber by regulating ROS homeostasis and energy metabolism.

Chilling injury is a major hindrance to cucumber fruit quality during cold storage. In this study, we evaluated the effects of fucoidan on fruit quality, reactive oxygen species homeostasis, and energy metabolism in cucumbers during cold storage. The results showed that, compared with the control cucumber fruit, fucoidan-treated cucumber fruit exhibited a lower chilling injury index and less weight loss, as well as reduced electrolyte leakage and malondialdehyde content. The most pronounced effects were observed following treatment with fucoidan at 15 g/L, which resulted in increased 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radical scavenging rates and reduced superoxide anion production rate and hydrogen peroxide content. The expression and activity levels of peroxidase, catalase, and superoxide dismutase were enhanced by fucoidan treatment. Further, fucoidan treatment maintained high levels of ascorbic acid and glutathione, and high ratios of ascorbic acid/dehydroascorbate and glutathione/oxidized glutathione. Moreover, fucoidan treatment increased the activities of ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, and glutathione reductase and their gene expression. Fucoidan treatment significantly delayed the decrease in ATP and ADP, while preventing an increase in AMP content. Finally, fucoidan treatment delayed the decrease of energy charge and the activities and gene expression of H+-ATPase, Ca2+-ATPase, cytochrome c oxidase, and succinate dehydrogenase in cucumber fruits. Altogether, our findings indicate that fucoidan can effectively enhance antioxidant capacity and maintain energy metabolism, thereby improving cucumber cold resistance during cold storage.