To investigate the effects of environmental stresses on ascorbic acid content and its redox status, the effects of freezing and drought on ascorbate and dehydro-ascorbate content and activities of four enzymes involve...To investigate the effects of environmental stresses on ascorbic acid content and its redox status, the effects of freezing and drought on ascorbate and dehydro-ascorbate content and activities of four enzymes involved in the ascorbate-glutathione cycle in some conifers were studied. The results showed that both freezing and drought induced the decrease in ascorbate content and the increase in dehydro-ascorbate content. The activities of ascorbate peroxidase (APX) and monodehydro-ascorbate reductase (MDAR) were decreased by freezing stress. At the beginning of exposure to air, water loss from detached needles induced the increase in the activities of APX and MDAR. Further water loss turned to decrease the APX and MDAR activities. The activities of dehydro-ascorbate reductase (DHAR) and glutathione reductase (GR) were not sensitive to changes in temperature and water content of the needles. It is concluded that moderate temperature or water stresses may induce the acclimation and increase in the ability of the H2O2 scavenging system, while strong stresses decrease the ability and induce injury of plant tissues. Correlation between ascorbate content and activities of related enzymes and cold tolerance of conifers were also reported.展开更多
[Objective] The research aimed to study the correlations between catalase(CAT) and ascorbate peroxidase(ASP) and the growth and development of rice roots under cadmium stress.[Method] Taking rice variety Zhonghua ...[Objective] The research aimed to study the correlations between catalase(CAT) and ascorbate peroxidase(ASP) and the growth and development of rice roots under cadmium stress.[Method] Taking rice variety Zhonghua No.11 as materials,the changes of rice seedlings under the treatment conditions of Cd,Cd+CAT inhibitor,Cd+APX inhibitor were studied.[Result] Under Cd stress,inhibition of CAT activity caused the significant inhibition on the growth of aerial parts,decreased the number of adventitious roots and lateral roots,but it can significant promote the elongation growth of adventitious roots and lateral roots.Moreover,the length of the first lateral root from root tip on the primary roots and adventitious roots was also increased than control.When APX activity was inhibited,the growth changes of rice were similar with that treated by CAT inhibitor.[Conclusion] CAT and APX may play important roles in the regulation of rice root system growth in both non-stress and Cd-stressed rice展开更多
As the most northerly mangrove species in China, Kandelia obovata may undergo extreme cold event stress. Enhancing the cold tolerance of this species is crucial to its successful afforestation. This study aimed to det...As the most northerly mangrove species in China, Kandelia obovata may undergo extreme cold event stress. Enhancing the cold tolerance of this species is crucial to its successful afforestation. This study aimed to determine the resistance of K. obovata seedlings to low temperature stress by cold acclimation and to explain the mechanisms for alleviating cold injury. To understand these mechanisms, seedlings that were acclimatized and not acclimatized were exposed to 5℃/- 2℃(day/night)for 48 h.Results showed that low temperature stress reduced leaf photosynthesis of non-acclimatized seedlings by inducing oxidative stress and structural damage to chloroplasts. These phenomena were shown by increasing levels of malondialdehyde (MDA), O2-and H2O2, as well as decreasing enzyme activities in the ascorbate–glutathione (AsA-GSH) cycle. However, cold-acclimatized seedlings had improved photosynthetic rates and efficiency of photosystem II (PSII) under low temperature stress. Compared with non-acclimatized seedlings, leaves of coldacclimatized seedlings under low temperature stress for 48 h exhibited higher anti-oxidative enzyme activities, lower levels of O2^- and H2O2, less damage to chloroplast structure, and removed 33.7% of MDA at low temperature stress for 48 h. The data indicate that cold acclimation enhances photosynthetic capacity by effectively regulating activation in the PSII electron transport and the AsA–GSH cycle to scavenge excess ROS in chloroplasts, while the latter is more important.展开更多
Ascorbate peroxidases (APX), localized in the cytosol, peroxisome, mitochondria, and chloroplasts of plant cells, catalyze the reduction of H2O2 to water by using ascorbic acid as the specific electron donor. To det...Ascorbate peroxidases (APX), localized in the cytosol, peroxisome, mitochondria, and chloroplasts of plant cells, catalyze the reduction of H2O2 to water by using ascorbic acid as the specific electron donor. To determine the role of peroxisomal type ascorbate peroxidase (pAPX), an antioxidant enzyme, in protection against salt-induced oxidative stress, transgenic Arabidopsis thaliana plant carrying a pAPX gene (HvAPX1) from barley (Hordeum vulgate L.) was analyzed. The transgenic line pAPX3 was found to be more tolerant to salt stress than the wild type. Irrespective of salt stress, there were no significant differences in Na^+, K^+, Ca^2+, and Mg^2+ contents and the ratio of K^+ to Na^+ between pAPX3 and the wild type. Clearly, the salt tolerance in pAPX3 was not due to the maintenance and reestablishment of cellular ion homeostasis. However, the degree of H2O2 and lipid peroxidation (measured as the levels of malondialdehyde) accumulation under salt stress was higher in the wild type than in pAPX3. The mechanism of salt tolerance in transgenic pAPX3 can thus be explained by reduction of oxidative stress injury. Under all conditions tested, activities of superoxide, glutathione reductase, and catalase were not significantly different between pAPX3 and the wild type. In contrast, the activity of APX was significantly higher in the transgenic plant than in wild type under salt stress. These results suggested that in higher plants, HvAPX1 played an important role in salt tolerance and was a candidate gene for developing salttolerant crop plants.展开更多
GDP-D-mannose pyrophosphorylase (GMPase) catalyses the synthesis of GDP-D-mannose and represents the first committed step in the synthesis of ascorbate. In the present study, the GMPase gene of tomato was introduced...GDP-D-mannose pyrophosphorylase (GMPase) catalyses the synthesis of GDP-D-mannose and represents the first committed step in the synthesis of ascorbate. In the present study, the GMPase gene of tomato was introduced into potato by Agrobacterium-mediated transformation. Two transgenic lines with higher GMPase expression were selected using qPCR and protein blot analyses. The results showed that the content of L-ascorbic acid (AsA) and the ratio of AsA/ DHA (dehydroascorbate) significantly increased in both leaves and tubers of transgenic potato plants. Both pigment content and photosynthetic rate were much higher in transgenic plants than in wild-type plants. Transgenic plants showed a distinguishable change in phenotype from the wild-type plants. Furthermore, transgenic plants showed delayed senescence.展开更多
Alfalfa (Medicago sativa L.) is an important forage crop in the world and it is of great signiifcance for the improvement of its salt tolerance. To improve salt tolerance in alfalfa, a rice ascorbate peroxidase gene...Alfalfa (Medicago sativa L.) is an important forage crop in the world and it is of great signiifcance for the improvement of its salt tolerance. To improve salt tolerance in alfalfa, a rice ascorbate peroxidase gene (OsAPX2) was introduced into alfalfa using Agrobacterium tumefaciens-mediated transformation with marker gene bar. The different T-DNA insertions in T1 transgenic alfalfa were identiifed by Southern hybridization. Three independent T2 transgenic lines were selected for stress analysis and the results showed that all of them were salt tolerant compared with wild-type plants. The transgenic plants had low levels of H2O2, malondialdehyde and relative electrical conductivity under salt and drought stresses. Moreover, the contents of chlorophyll and proline, and APX activity were high in transgenic plants under salt and drought stresses. Taken together, the overexpression of OsAPX2 enhances salt tolerance in alfalfa through scavenging reactive oxygen species.展开更多
Superoxide dismutase (SOD) and ascorbate peroxidase (APX) play central roles in the pathway for scavenging reactive oxygen species in plants, thereby contributing to the tolerance against abiotic stress. Here we repor...Superoxide dismutase (SOD) and ascorbate peroxidase (APX) play central roles in the pathway for scavenging reactive oxygen species in plants, thereby contributing to the tolerance against abiotic stress. Here we report the responses of cytosolic SOD (cSOD; sodCc1 and sodCc2) and cytosolic APX (cAPX; OsAPX1 and OsAPX2) genes to oxidative and abiotic stress in rice. RNA blot analyses revealed that methyl viologen treatment caused a more prominent induction of cAPXs compared with cSODs, and hydrogen peroxide treatment induced the expression of cAPXs whereas cSODs were not affected. These results suggest that cAPXs play more important roles in defense against oxidative stress compared with cSODs. It is noted that cSODs and cAPXs showed coordinate response to abscisic acid treatment which induced both sodCc1 and OsAPX2. However, cSODs and cAPXs responded differentially to drought, salt and chilling stress, which indicates that cSOD and cAPX genes are expressed differentially in response to oxidative and abiotic stress in rice.展开更多
Ascorbate peroxidase(APX) plays a key role in scavenging reactive oxygen species(ROS) in higher plants. However, there is very little information available on the APXs in kiwifruit(Actinidia), which is an economically...Ascorbate peroxidase(APX) plays a key role in scavenging reactive oxygen species(ROS) in higher plants. However, there is very little information available on the APXs in kiwifruit(Actinidia), which is an economically and nutritionally important horticultural crop with exceptionally high ascorbic acid(AsA) accumulation. This study aims to identify and characterize two cytosolic APX genes(AcAPX1 and AcAPX2) derived from A. chinensis ‘Hongyang’. The constitutive expression pattern was determined for both AcAPX1 and AcAPX2, and showed relatively higher expression abundances of AcAPX1 in leaf and AcAPX2 in root. Transcript levels of AcAPX1 and AcAPX2 were increased in kiwifruit roots treated with Na Cl. Subcellular localization assays using GFP-fusion proteins in Arabidopsis protoplasts showed that both AcAPX1 and AcAPX2 are targeted to the cytosol. Recombinant AcAPX1 or AcAPX2 proteins were successfully expressed in the prokaryotic expression system and their individual ascorbate peroxidase activities were determined. Finally, constitutive over-expression of AcAPX1 or AcAPX2 could dramatically increase total As A, glutathione level and salinity tolerance under Na Cl stress in Arabidopsis thaliana. Our findings revealed that cytosolic AcAPX1/2 may play an important protective role in the responses to unfavorable environmental stimuli in kiwifruit.展开更多
An antitumour complex,cis-diammineplatinum(Ⅱ)ascorbate,has been synthesized and its X-ray structure is given.Quantum chemistry calculation compared with its diaminecyclohexane derivatives indicates that they have dif...An antitumour complex,cis-diammineplatinum(Ⅱ)ascorbate,has been synthesized and its X-ray structure is given.Quantum chemistry calculation compared with its diaminecyclohexane derivatives indicates that they have different electron distribution.展开更多
Investigation of male sterility mutations is an effective approach for identification of genes involved in anther and pollen development. The comparison of “cytological phenotypes” of newly induced mutants with phen...Investigation of male sterility mutations is an effective approach for identification of genes involved in anther and pollen development. The comparison of “cytological phenotypes” of newly induced mutants with phenotypes determined by already known genes favors elucidation of genetic control of diverse microsporo- and gametogenesis stages. In this paper, we describe pollen development in the grain sorghum line Zh10-asc1 with mutation of male sterility. This line was obtained from callus culture treated by sodium ascorbate. A wide spectrum of abnormalities in microsporogenesis have been found, such as cytomixis, chromosomal laggards, chromosome disjunction, adhesion of chromosomes, disturbed cytokinesis, and others. In tapetum, the cells with one nucleus, with unequal nuclei, and with micronuclei have been observed. During pollen grain (PG) maturation abnormalities in starch accumulation and delay of development often took place. In mature anthers, a variety of pollen grain types have been revealed: fertile, of irregular shape, incompletely filled with starch, PGs delayed at the uni-nucleate or bi-nucleate gametophyte stages, with partially or fully degenerated contents, and with abnormal coloration. Variation in spectrum and the frequency of disturbances between the flowers of one and the same plant have been revealed. The reasons for significant genetic and epigenetic instability are discussed.展开更多
Arsenic Trioxide (ATO) is widely acknowledged as the treatment of choice for Acute Promyelocytic Leukemia (APL). It is a “two-sided” drug since it can induce differentiation or kill APL and other tumor cells accordi...Arsenic Trioxide (ATO) is widely acknowledged as the treatment of choice for Acute Promyelocytic Leukemia (APL). It is a “two-sided” drug since it can induce differentiation or kill APL and other tumor cells according to the dosage. Part of the cytotoxic effects of ATO on APL cells is due to its pro-oxidant activity, a characteristic which ATO shares with a number of other compounds, including high doses of ascorbate (ASC). In a comparative investigation on the cytotoxic effects of both ATO and ASC on HL60 (APL) cell lines, in Vitro, we have been able to confirm the known cytotoxic effects of ATO, but, more importantly, we have demonstrated that ASC is significantly more effective than ATO, in killing these cancer cells in Vitro, when the concentrations are maintained within the millimolar (mM) range, i.e. the range of plasma concentrations at which ASC induces oxidative damage to tumor cells. Since these plasma levels can be reached only by the intravenous administration of high doses of ASC, we propose that intravenous high doses of ASC may represent a potentially revolutionary new approach in the management of APL.展开更多
All genes encoding chloroplast antioxidant enzymes are nuclear-encoded and posttranscriptionally targeted to chloroplasts. The transcript levels of most of them decreased upon sucrose feeding like the transcript level...All genes encoding chloroplast antioxidant enzymes are nuclear-encoded and posttranscriptionally targeted to chloroplasts. The transcript levels of most of them decreased upon sucrose feeding like the transcript levels of many genes encoding components of the photosynthetic electron transport chain. However, the transcript abundance of stromal ascorbate peroxidase (s-APX; At4g08390) increased. Due to mild sugar application conditions, the plants kept the phosphorylation status of the ADP+ATP pool and the redox states of the NADPH+NADP~ and the ascorbate pools under control, which excludes them as signals in s-APX regulation. Correlation with ascorbate pool size regulation and comparison of transcript abundance regulation in the starch-biosynthetic mutant adgl, the ascorbate biosynthesis mutant vtcl, and the abscisic acid (ABA) biosynthetic mutant aba2 showed a link between sugar induction of s-APX and ascorbate biosynthesis.展开更多
The possible association with nitric oxide (NO) and ascorbate-glutathione (AsA-GSH) cycle in regulating aluminum (Al) tolerance of wheat (Triticum aestivum L.) was investigated using two genotypes with differe...The possible association with nitric oxide (NO) and ascorbate-glutathione (AsA-GSH) cycle in regulating aluminum (Al) tolerance of wheat (Triticum aestivum L.) was investigated using two genotypes with different Al resistance. Exposure to Al inhibited root elongation, and triggered lipid peroxidation and oxidation of AsA to dehydroascorbate and GSH to glutathione disulfide in wheat roots. Exogenous NO significantly increased endogenous NO levels, and subsequently al eviated Al-induced inhibition of root elongation and oxidation of AsA and GSH to maintain the redox molecules in the reduced form in both wheat genotypes. Under Al stress, significantly increased activities and gene transcriptional levels of ascorbate peroxi-dase, glutathione reductase, and dehydroascorbate reductase, were observed in the root tips of the Al-tolerant genotype Jian-864. Nitric oxide application enhanced the activity and gene transcriptional level of these enzymes in both wheat geno-types. g-Glutamylcysteine synthetase was not significantly affected by Al or NO, but NO treatments increased the activity of glutathione peroxidase and glutathione S-transferase to a greater extent than the Al-treated wheat seedlings. Proline was significantly decreased by Al, while it was not affected by NO. These results clearly suggest that NO protects wheat root against Al-induced oxidative stress, possibly through its regulation of the AsA-GSH cycle.展开更多
The sensitivity of Chinese soybean cultivars to ambient ozone(O3) in the field is unknown,although soybean is a major staple food in China. Using ethylenediurea(EDU) as an O3 protectant, we tested the gas exchange...The sensitivity of Chinese soybean cultivars to ambient ozone(O3) in the field is unknown,although soybean is a major staple food in China. Using ethylenediurea(EDU) as an O3 protectant, we tested the gas exchange, pigments, antioxidants and biomass of 19 cultivars exposed to 28 ppm·hr AOT40(accumulated O3 over an hourly concentration threshold of40 ppb) over the growing season at a field site in China. By comparing the average biomass with and without EDU, we estimated the cultivar-specific sensitivity to O3 and ranked the cultivars from very tolerant(〈 10% change) to highly sensitive(〉 45% change), which helps in choosing the best-suited cultivars for local cultivation. Higher lipid peroxidation and activity of the ascorbate peroxidase enzyme were major responses to O3 damage, which eventually translated into lower biomass production. The constitutional level of total ascorbate in the leaves was the most important parameter explaining O3 sensitivity among these cultivars. Surprisingly, the role of stomatal conductance was insignificant. These results will guide future breeding efforts towards more O3-tolerant cultivars in China, while strategies for implementing control measures of regional O3 pollution are being implemented. Overall, these results suggest that present ambient O3 pollution is a serious concern for soybean in China, which highlights the urgent need for policy-making actions to protect this critical staple food.展开更多
To investigate the sensitive site of antioxidant systems in chloroplast under cadmium stress and its consequence on reactive oxygen species production and action, the sub-organellar localization of chloroplast superox...To investigate the sensitive site of antioxidant systems in chloroplast under cadmium stress and its consequence on reactive oxygen species production and action, the sub-organellar localization of chloroplast superoxide dismutases (SOD, EC 1.15.1.1) and ascorbic peroxidase (APX, EC 1.11.1.11) isoenzymes and changes of enzymes activities under cadmium stress were investigated in tomato seedlings. Two APX isoforms, one thylakoid-bound and one stromal, were detected. Cd at 50 μM induced a moderate increase of SOD activities but a rapid inactivation of both APX isoenzymes. APX inactivation was mainly related to the decrease of ascorbate concentration, as supported by in vitro treatment of exogenous ascorbate and APX kinetic properties under Cd stress. H2O2 accumulation in chloroplast, as a consequence of APX inactivation, was associated with a 60% loss of Rubisco (EC 4.1.1.39) activity, which could be partially accounted for by a 10% loss of Rubisco content. Protein oxidation assay found that the Rubisco large subunit was the most prominent carbonylated protein; the level of carbonylated Rubisco large subunit increased fivefold after Cd exposure. Thiol groups in the Rubisco large subunit were oxidized, as indicated by non-reducing electrophoresis. Treating crude extract with H2O2 resulted in a similar pattern of protein oxidation and thiols oxidation with that observed in Cd-treated plants. Our study indicates that APXs in the chloroplast is a highly sensitive site of antioxidant systems under Cd stress, and the inactivation of APX could be mainly responsible for oxidative modification to Rubisco and subsequent decrease in its activity.展开更多
Apoplastic ascorbate oxidases(AOs)play a critical role in reactive oxygen species(RoS)-mediated innate host immunity by regulating the apoplast redox state.To date,little is known about how apoplastic effectors of the...Apoplastic ascorbate oxidases(AOs)play a critical role in reactive oxygen species(RoS)-mediated innate host immunity by regulating the apoplast redox state.To date,little is known about how apoplastic effectors of the riceblast fungus Magnaportheoryzaemodulate the apoplast redox state of rice to subvert plant immunity.In this study,we demonstrated that M.oryzae MoAo1 is an Ao that plays a role in virulence by modulating the apoplast redox status of rice cells.We showed that MoAo1 inhibits the activity of rice OsAO3and OsAO4,which also regulate the apoplast redox status and plant immunity.In addition,we found that MoAo1,OsAO3,andOsAO4 allexhibit polymorphic variations whosevaried interactions orchestrate pathogen virulence and rice immunity.Taken together,our results reveal a critical role for extracellular redox enzymes during rice blast infection and shed light on the importance of the apoplast redox state anditsregulation inplant-pathogeninteractions.展开更多
Southern corn leaf blight (SCLB), caused by Bipolarismaydis, is one of the most devastatingdiseases affecting maize production. However,only one SLCB resistance gene, conferring partialresistance, is currently known, ...Southern corn leaf blight (SCLB), caused by Bipolarismaydis, is one of the most devastatingdiseases affecting maize production. However,only one SLCB resistance gene, conferring partialresistance, is currently known, underscoring theimportance of isolating new SCLB resistancerelatedgenes. Here, we performed a comparativeproteomic analysis and identified 258 proteinsshowing differential abundance during the maizeresponse to B. maydis. These proteins included anascorbate peroxidase (Zea mays ascorbate peroxidase1 (ZmAPX1)) encoded by a gene locatedwithin the mapping interval of a previously identifiedquantitative trait locus associated with SCLBresistance. ZmAPX1 overexpression resulted inlower H_(2)O_(2) accumulation and enhanced resistanceagainst B. maydis. Jasmonic acid (JA)contents and transcript levels for JA biosynthesisand responsive genes increased in ZmAPX1-overexpressing plants infected with B. maydis,whereas Zmapx1 mutants showed the oppositeeffects. We further determined that low levels of H_(2)O_(2) are accompanied by an accumulation of JAthat enhances SCLB resistance. These resultsdemonstrate that ZmAPX1 positively regulatesSCLB resistance by decreasing H_(2)O_(2) accumulationand activating the JA-mediated defensesignaling pathway. This study identified ZmAPX1as a potentially useful gene for increasing SCLBresistance. Furthermore, the generated datamay be relevant for clarifying the functions ofplant APXs.展开更多
Plants encounter a variety of stresses in natural environments. One-year-old pot-grown trees of pear(Pyrus pyrifolia Nakai cv. Cuiguan and Wonhwang) were exposed to two heat stress regimes. Under constant short-term h...Plants encounter a variety of stresses in natural environments. One-year-old pot-grown trees of pear(Pyrus pyrifolia Nakai cv. Cuiguan and Wonhwang) were exposed to two heat stress regimes. Under constant short-term heat stress, chloroplasts and mitochondria were visibly damaged. Relative chlorophyll content and maximum photochemical efficiency of photosystem II were significantly decreased, which indicated that the leaf photosynthetic capability declined. Under chronic heat stress, mesophyll cell ultrastructure was not obviously damaged, but leaf photosynthetic capability was still restrained. As chronic heat stress was a simulation of the natural environment in summer, further study of the responses under this stress regime was undertaken. Ascorbate peroxidase(APX) activity was increased in ‘Cuiguan', but not in ‘Wonhwang'. Inducible expression of PpAPX genes in the cytoplasm, chloroplasts and peroxisomes was consistent with increased APX activity in ‘Cuiguan', whereas only weak induction of PpAPX genes was observed in ‘Wonhwang'. The isoenzymes cytosolic APX1(cAPX1) and stromal APX(sAPX) were confirmed to be localized in the cytoplasm and chloroplasts, respectively.展开更多
Ascorbate peroxidase, a haem protein (EC 1.11.1.11), efficiently scavenges hydrogen peroxide (H2O2) in cytosol and chloroplasts of plants. In this study, a fulllength coding sequence of thylakoid-bound ascorbate p...Ascorbate peroxidase, a haem protein (EC 1.11.1.11), efficiently scavenges hydrogen peroxide (H2O2) in cytosol and chloroplasts of plants. In this study, a fulllength coding sequence of thylakoid-bound ascorbate peroxidase cDNA (TatAPX) was cloned from a drought tolerant wheat cultivar C306. Homology modeling of the TatAPX protein was performed by using the template crystal structure of chloroplastic ascorbate peroxidase from tobacco plant (PDB: IIYN). The model structure was further refined by molecular mechanics and dynamic methods using various tools such as PROCHECK, ProSA and Verify3D. The predicted model was then tested for docking with H2O2, the substrate for TatAPX enzyme. The results revealed that Arg233 and Glu255 in the predicted active site of the enzyme are two important amino acid residues responsible for strong hydrogen bonding affinity with H2O2, which might play an important role in scavenging of H2O2 from the plant system.展开更多
The vitamin C(VC) in crops was successfully determined using ascorbate oxidase(AO) electrochemical biosensor based on the biocompatible poly(3,4-ethylenedioxythiophene)(PEDOT) matrices,which was easily prepare...The vitamin C(VC) in crops was successfully determined using ascorbate oxidase(AO) electrochemical biosensor based on the biocompatible poly(3,4-ethylenedioxythiophene)(PEDOT) matrices,which was easily prepared by one-step electrodeposition technique in ionic liquid microemulsions.The fabricated biosensor displayed excellent bioelectrocatalytic performance to the oxidation of VC,wide linear range,low detection limit,fast response time,good operational and storage stability,the good results of the determination of VC in vegetable crops indicated that the fabricated biosensor will be a good candidate for the physiological and biochemical studies of crops in near future.展开更多
文摘To investigate the effects of environmental stresses on ascorbic acid content and its redox status, the effects of freezing and drought on ascorbate and dehydro-ascorbate content and activities of four enzymes involved in the ascorbate-glutathione cycle in some conifers were studied. The results showed that both freezing and drought induced the decrease in ascorbate content and the increase in dehydro-ascorbate content. The activities of ascorbate peroxidase (APX) and monodehydro-ascorbate reductase (MDAR) were decreased by freezing stress. At the beginning of exposure to air, water loss from detached needles induced the increase in the activities of APX and MDAR. Further water loss turned to decrease the APX and MDAR activities. The activities of dehydro-ascorbate reductase (DHAR) and glutathione reductase (GR) were not sensitive to changes in temperature and water content of the needles. It is concluded that moderate temperature or water stresses may induce the acclimation and increase in the ability of the H2O2 scavenging system, while strong stresses decrease the ability and induce injury of plant tissues. Correlation between ascorbate content and activities of related enzymes and cold tolerance of conifers were also reported.
基金Supported by National Natural Science Foundation of China(30671126)~~
文摘[Objective] The research aimed to study the correlations between catalase(CAT) and ascorbate peroxidase(ASP) and the growth and development of rice roots under cadmium stress.[Method] Taking rice variety Zhonghua No.11 as materials,the changes of rice seedlings under the treatment conditions of Cd,Cd+CAT inhibitor,Cd+APX inhibitor were studied.[Result] Under Cd stress,inhibition of CAT activity caused the significant inhibition on the growth of aerial parts,decreased the number of adventitious roots and lateral roots,but it can significant promote the elongation growth of adventitious roots and lateral roots.Moreover,the length of the first lateral root from root tip on the primary roots and adventitious roots was also increased than control.When APX activity was inhibited,the growth changes of rice were similar with that treated by CAT inhibitor.[Conclusion] CAT and APX may play important roles in the regulation of rice root system growth in both non-stress and Cd-stressed rice
基金supported by Zhejiang Provincial Natural Science Foundation of China(Grant Nos.LY18C030001 and LQ13C030002)National Natural Science Foundation of China(Grant No.41776097)+4 种基金Special Funding for Research of National Oceanic Public Service Industry of China(Grant No.201505028)National Science and Technology Basic Resources Survey Special of China(Grant No.2017FY100700)Zhejiang Province Science and Technology Plan Project of China(Grant Nos.2013C25096 and2014F50003)Zhejiang Province Foundation of the Nonprofit Technology Research Projects of China(Grant No.2015C33227)Wenzhou Municipal Science and Technology Plan Project of China(Grant Nos.N20140046,N20170008 and S20160004)
文摘As the most northerly mangrove species in China, Kandelia obovata may undergo extreme cold event stress. Enhancing the cold tolerance of this species is crucial to its successful afforestation. This study aimed to determine the resistance of K. obovata seedlings to low temperature stress by cold acclimation and to explain the mechanisms for alleviating cold injury. To understand these mechanisms, seedlings that were acclimatized and not acclimatized were exposed to 5℃/- 2℃(day/night)for 48 h.Results showed that low temperature stress reduced leaf photosynthesis of non-acclimatized seedlings by inducing oxidative stress and structural damage to chloroplasts. These phenomena were shown by increasing levels of malondialdehyde (MDA), O2-and H2O2, as well as decreasing enzyme activities in the ascorbate–glutathione (AsA-GSH) cycle. However, cold-acclimatized seedlings had improved photosynthetic rates and efficiency of photosystem II (PSII) under low temperature stress. Compared with non-acclimatized seedlings, leaves of coldacclimatized seedlings under low temperature stress for 48 h exhibited higher anti-oxidative enzyme activities, lower levels of O2^- and H2O2, less damage to chloroplast structure, and removed 33.7% of MDA at low temperature stress for 48 h. The data indicate that cold acclimation enhances photosynthetic capacity by effectively regulating activation in the PSII electron transport and the AsA–GSH cycle to scavenge excess ROS in chloroplasts, while the latter is more important.
基金a grant of CAS (Chinese Academy of Sciences) Research Program on Soil Biosystems and Agro-Product Safety (No.CXTD-Z2005-4)the Knowledge Innovation Project of CAS (No.KZCX3-SW-439).
文摘Ascorbate peroxidases (APX), localized in the cytosol, peroxisome, mitochondria, and chloroplasts of plant cells, catalyze the reduction of H2O2 to water by using ascorbic acid as the specific electron donor. To determine the role of peroxisomal type ascorbate peroxidase (pAPX), an antioxidant enzyme, in protection against salt-induced oxidative stress, transgenic Arabidopsis thaliana plant carrying a pAPX gene (HvAPX1) from barley (Hordeum vulgate L.) was analyzed. The transgenic line pAPX3 was found to be more tolerant to salt stress than the wild type. Irrespective of salt stress, there were no significant differences in Na^+, K^+, Ca^2+, and Mg^2+ contents and the ratio of K^+ to Na^+ between pAPX3 and the wild type. Clearly, the salt tolerance in pAPX3 was not due to the maintenance and reestablishment of cellular ion homeostasis. However, the degree of H2O2 and lipid peroxidation (measured as the levels of malondialdehyde) accumulation under salt stress was higher in the wild type than in pAPX3. The mechanism of salt tolerance in transgenic pAPX3 can thus be explained by reduction of oxidative stress injury. Under all conditions tested, activities of superoxide, glutathione reductase, and catalase were not significantly different between pAPX3 and the wild type. In contrast, the activity of APX was significantly higher in the transgenic plant than in wild type under salt stress. These results suggested that in higher plants, HvAPX1 played an important role in salt tolerance and was a candidate gene for developing salttolerant crop plants.
基金supported by the National Basic Re-search Program of China (2009CB119000)the National Natural Science Foundation of China (30771473)funded by Key Laboratory of Horticultural Crops Ge-netic Improvement, Ministry of Agriculture of China
文摘GDP-D-mannose pyrophosphorylase (GMPase) catalyses the synthesis of GDP-D-mannose and represents the first committed step in the synthesis of ascorbate. In the present study, the GMPase gene of tomato was introduced into potato by Agrobacterium-mediated transformation. Two transgenic lines with higher GMPase expression were selected using qPCR and protein blot analyses. The results showed that the content of L-ascorbic acid (AsA) and the ratio of AsA/ DHA (dehydroascorbate) significantly increased in both leaves and tubers of transgenic potato plants. Both pigment content and photosynthetic rate were much higher in transgenic plants than in wild-type plants. Transgenic plants showed a distinguishable change in phenotype from the wild-type plants. Furthermore, transgenic plants showed delayed senescence.
基金supported by the National 973 Program of China (2014CB138700)
文摘Alfalfa (Medicago sativa L.) is an important forage crop in the world and it is of great signiifcance for the improvement of its salt tolerance. To improve salt tolerance in alfalfa, a rice ascorbate peroxidase gene (OsAPX2) was introduced into alfalfa using Agrobacterium tumefaciens-mediated transformation with marker gene bar. The different T-DNA insertions in T1 transgenic alfalfa were identiifed by Southern hybridization. Three independent T2 transgenic lines were selected for stress analysis and the results showed that all of them were salt tolerant compared with wild-type plants. The transgenic plants had low levels of H2O2, malondialdehyde and relative electrical conductivity under salt and drought stresses. Moreover, the contents of chlorophyll and proline, and APX activity were high in transgenic plants under salt and drought stresses. Taken together, the overexpression of OsAPX2 enhances salt tolerance in alfalfa through scavenging reactive oxygen species.
基金supported by the Grants-in-Aid for Scientific Research (Grant No. 10460149 to K.T. and Grant No. 11740448 to S.M.) from the Ministry of Education, Culture, Sports, Science and Technology of Japana grant from the Rice Genome Research Program (Grant No. MP2106 to K.T.) from the Ministry of Agriculture, Forestry and Fisheries of Japan
文摘Superoxide dismutase (SOD) and ascorbate peroxidase (APX) play central roles in the pathway for scavenging reactive oxygen species in plants, thereby contributing to the tolerance against abiotic stress. Here we report the responses of cytosolic SOD (cSOD; sodCc1 and sodCc2) and cytosolic APX (cAPX; OsAPX1 and OsAPX2) genes to oxidative and abiotic stress in rice. RNA blot analyses revealed that methyl viologen treatment caused a more prominent induction of cAPXs compared with cSODs, and hydrogen peroxide treatment induced the expression of cAPXs whereas cSODs were not affected. These results suggest that cAPXs play more important roles in defense against oxidative stress compared with cSODs. It is noted that cSODs and cAPXs showed coordinate response to abscisic acid treatment which induced both sodCc1 and OsAPX2. However, cSODs and cAPXs responded differentially to drought, salt and chilling stress, which indicates that cSOD and cAPX genes are expressed differentially in response to oxidative and abiotic stress in rice.
基金funded by the National Natural Science Foundation of China (31972474)the Natural Science Research Program of Universities of Anhui Province, China (K1832004)+2 种基金the Leading Talent Group Funding of Anhui Province, China (WRMR-2020-75)the Natural Science Foundation of Anhui Province, China (19232002)the Anhui Agriculture University Shennong Scholar Project, China (RC321901)。
文摘Ascorbate peroxidase(APX) plays a key role in scavenging reactive oxygen species(ROS) in higher plants. However, there is very little information available on the APXs in kiwifruit(Actinidia), which is an economically and nutritionally important horticultural crop with exceptionally high ascorbic acid(AsA) accumulation. This study aims to identify and characterize two cytosolic APX genes(AcAPX1 and AcAPX2) derived from A. chinensis ‘Hongyang’. The constitutive expression pattern was determined for both AcAPX1 and AcAPX2, and showed relatively higher expression abundances of AcAPX1 in leaf and AcAPX2 in root. Transcript levels of AcAPX1 and AcAPX2 were increased in kiwifruit roots treated with Na Cl. Subcellular localization assays using GFP-fusion proteins in Arabidopsis protoplasts showed that both AcAPX1 and AcAPX2 are targeted to the cytosol. Recombinant AcAPX1 or AcAPX2 proteins were successfully expressed in the prokaryotic expression system and their individual ascorbate peroxidase activities were determined. Finally, constitutive over-expression of AcAPX1 or AcAPX2 could dramatically increase total As A, glutathione level and salinity tolerance under Na Cl stress in Arabidopsis thaliana. Our findings revealed that cytosolic AcAPX1/2 may play an important protective role in the responses to unfavorable environmental stimuli in kiwifruit.
基金suported by the Medicine and Drug Bureau of China
文摘An antitumour complex,cis-diammineplatinum(Ⅱ)ascorbate,has been synthesized and its X-ray structure is given.Quantum chemistry calculation compared with its diaminecyclohexane derivatives indicates that they have different electron distribution.
文摘Investigation of male sterility mutations is an effective approach for identification of genes involved in anther and pollen development. The comparison of “cytological phenotypes” of newly induced mutants with phenotypes determined by already known genes favors elucidation of genetic control of diverse microsporo- and gametogenesis stages. In this paper, we describe pollen development in the grain sorghum line Zh10-asc1 with mutation of male sterility. This line was obtained from callus culture treated by sodium ascorbate. A wide spectrum of abnormalities in microsporogenesis have been found, such as cytomixis, chromosomal laggards, chromosome disjunction, adhesion of chromosomes, disturbed cytokinesis, and others. In tapetum, the cells with one nucleus, with unequal nuclei, and with micronuclei have been observed. During pollen grain (PG) maturation abnormalities in starch accumulation and delay of development often took place. In mature anthers, a variety of pollen grain types have been revealed: fertile, of irregular shape, incompletely filled with starch, PGs delayed at the uni-nucleate or bi-nucleate gametophyte stages, with partially or fully degenerated contents, and with abnormal coloration. Variation in spectrum and the frequency of disturbances between the flowers of one and the same plant have been revealed. The reasons for significant genetic and epigenetic instability are discussed.
文摘Arsenic Trioxide (ATO) is widely acknowledged as the treatment of choice for Acute Promyelocytic Leukemia (APL). It is a “two-sided” drug since it can induce differentiation or kill APL and other tumor cells according to the dosage. Part of the cytotoxic effects of ATO on APL cells is due to its pro-oxidant activity, a characteristic which ATO shares with a number of other compounds, including high doses of ascorbate (ASC). In a comparative investigation on the cytotoxic effects of both ATO and ASC on HL60 (APL) cell lines, in Vitro, we have been able to confirm the known cytotoxic effects of ATO, but, more importantly, we have demonstrated that ASC is significantly more effective than ATO, in killing these cancer cells in Vitro, when the concentrations are maintained within the millimolar (mM) range, i.e. the range of plasma concentrations at which ASC induces oxidative damage to tumor cells. Since these plasma levels can be reached only by the intravenous administration of high doses of ASC, we propose that intravenous high doses of ASC may represent a potentially revolutionary new approach in the management of APL.
文摘All genes encoding chloroplast antioxidant enzymes are nuclear-encoded and posttranscriptionally targeted to chloroplasts. The transcript levels of most of them decreased upon sucrose feeding like the transcript levels of many genes encoding components of the photosynthetic electron transport chain. However, the transcript abundance of stromal ascorbate peroxidase (s-APX; At4g08390) increased. Due to mild sugar application conditions, the plants kept the phosphorylation status of the ADP+ATP pool and the redox states of the NADPH+NADP~ and the ascorbate pools under control, which excludes them as signals in s-APX regulation. Correlation with ascorbate pool size regulation and comparison of transcript abundance regulation in the starch-biosynthetic mutant adgl, the ascorbate biosynthesis mutant vtcl, and the abscisic acid (ABA) biosynthetic mutant aba2 showed a link between sugar induction of s-APX and ascorbate biosynthesis.
基金supported by the National Basic Research Program (973 Program) of China (2013CB127403)National Natural Science Foundation of China (31272237,30771292)+1 种基金the Foundation for University Ph.D.Granting Disciplines of the Ministry of Education (20120101110130)IPNI
文摘The possible association with nitric oxide (NO) and ascorbate-glutathione (AsA-GSH) cycle in regulating aluminum (Al) tolerance of wheat (Triticum aestivum L.) was investigated using two genotypes with different Al resistance. Exposure to Al inhibited root elongation, and triggered lipid peroxidation and oxidation of AsA to dehydroascorbate and GSH to glutathione disulfide in wheat roots. Exogenous NO significantly increased endogenous NO levels, and subsequently al eviated Al-induced inhibition of root elongation and oxidation of AsA and GSH to maintain the redox molecules in the reduced form in both wheat genotypes. Under Al stress, significantly increased activities and gene transcriptional levels of ascorbate peroxi-dase, glutathione reductase, and dehydroascorbate reductase, were observed in the root tips of the Al-tolerant genotype Jian-864. Nitric oxide application enhanced the activity and gene transcriptional level of these enzymes in both wheat geno-types. g-Glutamylcysteine synthetase was not significantly affected by Al or NO, but NO treatments increased the activity of glutathione peroxidase and glutathione S-transferase to a greater extent than the Al-treated wheat seedlings. Proline was significantly decreased by Al, while it was not affected by NO. These results clearly suggest that NO protects wheat root against Al-induced oxidative stress, possibly through its regulation of the AsA-GSH cycle.
基金supported by State Key Laboratory of Soil and Sustainable Agriculture(No.Y20160030)the Hundred Talents Program,Chinese Academy of Sciences,Chinese Academy of Sciences President's International Fellowship Initiative(PIFI)for Senior Scientists(Grant Number 2016VBA057)CNR-CAS bilateral agreement 2017–2019(Ozone impacts on plant ecosystems in China and Italy)
文摘The sensitivity of Chinese soybean cultivars to ambient ozone(O3) in the field is unknown,although soybean is a major staple food in China. Using ethylenediurea(EDU) as an O3 protectant, we tested the gas exchange, pigments, antioxidants and biomass of 19 cultivars exposed to 28 ppm·hr AOT40(accumulated O3 over an hourly concentration threshold of40 ppb) over the growing season at a field site in China. By comparing the average biomass with and without EDU, we estimated the cultivar-specific sensitivity to O3 and ranked the cultivars from very tolerant(〈 10% change) to highly sensitive(〉 45% change), which helps in choosing the best-suited cultivars for local cultivation. Higher lipid peroxidation and activity of the ascorbate peroxidase enzyme were major responses to O3 damage, which eventually translated into lower biomass production. The constitutional level of total ascorbate in the leaves was the most important parameter explaining O3 sensitivity among these cultivars. Surprisingly, the role of stomatal conductance was insignificant. These results will guide future breeding efforts towards more O3-tolerant cultivars in China, while strategies for implementing control measures of regional O3 pollution are being implemented. Overall, these results suggest that present ambient O3 pollution is a serious concern for soybean in China, which highlights the urgent need for policy-making actions to protect this critical staple food.
基金the National Natural Science Foundation of China (30671471and 30571296)
文摘To investigate the sensitive site of antioxidant systems in chloroplast under cadmium stress and its consequence on reactive oxygen species production and action, the sub-organellar localization of chloroplast superoxide dismutases (SOD, EC 1.15.1.1) and ascorbic peroxidase (APX, EC 1.11.1.11) isoenzymes and changes of enzymes activities under cadmium stress were investigated in tomato seedlings. Two APX isoforms, one thylakoid-bound and one stromal, were detected. Cd at 50 μM induced a moderate increase of SOD activities but a rapid inactivation of both APX isoenzymes. APX inactivation was mainly related to the decrease of ascorbate concentration, as supported by in vitro treatment of exogenous ascorbate and APX kinetic properties under Cd stress. H2O2 accumulation in chloroplast, as a consequence of APX inactivation, was associated with a 60% loss of Rubisco (EC 4.1.1.39) activity, which could be partially accounted for by a 10% loss of Rubisco content. Protein oxidation assay found that the Rubisco large subunit was the most prominent carbonylated protein; the level of carbonylated Rubisco large subunit increased fivefold after Cd exposure. Thiol groups in the Rubisco large subunit were oxidized, as indicated by non-reducing electrophoresis. Treating crude extract with H2O2 resulted in a similar pattern of protein oxidation and thiols oxidation with that observed in Cd-treated plants. Our study indicates that APXs in the chloroplast is a highly sensitive site of antioxidant systems under Cd stress, and the inactivation of APX could be mainly responsible for oxidative modification to Rubisco and subsequent decrease in its activity.
基金This research was supported by the China National Funds for Innovative Research Groups(31721004)the key program of the Natural Science Foundation of China(NSFC)(32030091)+3 种基金the NSFC Youth Program(31901832)Youth Program for Natural Science Foundation of Jiangsu Province(BK2019054)the program of NSFC-DFG(31861133017)the NSFC program(32172377)。
文摘Apoplastic ascorbate oxidases(AOs)play a critical role in reactive oxygen species(RoS)-mediated innate host immunity by regulating the apoplast redox state.To date,little is known about how apoplastic effectors of the riceblast fungus Magnaportheoryzaemodulate the apoplast redox state of rice to subvert plant immunity.In this study,we demonstrated that M.oryzae MoAo1 is an Ao that plays a role in virulence by modulating the apoplast redox status of rice cells.We showed that MoAo1 inhibits the activity of rice OsAO3and OsAO4,which also regulate the apoplast redox status and plant immunity.In addition,we found that MoAo1,OsAO3,andOsAO4 allexhibit polymorphic variations whosevaried interactions orchestrate pathogen virulence and rice immunity.Taken together,our results reveal a critical role for extracellular redox enzymes during rice blast infection and shed light on the importance of the apoplast redox state anditsregulation inplant-pathogeninteractions.
基金supported by the National Natural Science Foundation of China (31872872 and U1804113)the National Key Research and Development Program of China (2016YFD0101003)
文摘Southern corn leaf blight (SCLB), caused by Bipolarismaydis, is one of the most devastatingdiseases affecting maize production. However,only one SLCB resistance gene, conferring partialresistance, is currently known, underscoring theimportance of isolating new SCLB resistancerelatedgenes. Here, we performed a comparativeproteomic analysis and identified 258 proteinsshowing differential abundance during the maizeresponse to B. maydis. These proteins included anascorbate peroxidase (Zea mays ascorbate peroxidase1 (ZmAPX1)) encoded by a gene locatedwithin the mapping interval of a previously identifiedquantitative trait locus associated with SCLBresistance. ZmAPX1 overexpression resulted inlower H_(2)O_(2) accumulation and enhanced resistanceagainst B. maydis. Jasmonic acid (JA)contents and transcript levels for JA biosynthesisand responsive genes increased in ZmAPX1-overexpressing plants infected with B. maydis,whereas Zmapx1 mutants showed the oppositeeffects. We further determined that low levels of H_(2)O_(2) are accompanied by an accumulation of JAthat enhances SCLB resistance. These resultsdemonstrate that ZmAPX1 positively regulatesSCLB resistance by decreasing H_(2)O_(2) accumulationand activating the JA-mediated defensesignaling pathway. This study identified ZmAPX1as a potentially useful gene for increasing SCLBresistance. Furthermore, the generated datamay be relevant for clarifying the functions ofplant APXs.
基金supported by the Earmarked Fund for the Modern Agro-industry Technology Research System,China
文摘Plants encounter a variety of stresses in natural environments. One-year-old pot-grown trees of pear(Pyrus pyrifolia Nakai cv. Cuiguan and Wonhwang) were exposed to two heat stress regimes. Under constant short-term heat stress, chloroplasts and mitochondria were visibly damaged. Relative chlorophyll content and maximum photochemical efficiency of photosystem II were significantly decreased, which indicated that the leaf photosynthetic capability declined. Under chronic heat stress, mesophyll cell ultrastructure was not obviously damaged, but leaf photosynthetic capability was still restrained. As chronic heat stress was a simulation of the natural environment in summer, further study of the responses under this stress regime was undertaken. Ascorbate peroxidase(APX) activity was increased in ‘Cuiguan', but not in ‘Wonhwang'. Inducible expression of PpAPX genes in the cytoplasm, chloroplasts and peroxisomes was consistent with increased APX activity in ‘Cuiguan', whereas only weak induction of PpAPX genes was observed in ‘Wonhwang'. The isoenzymes cytosolic APX1(cAPX1) and stromal APX(sAPX) were confirmed to be localized in the cytoplasm and chloroplasts, respectively.
基金supported by the Indian Council of Agricultural Research (ICAR)sponsored Network Project on Transgenics in Crops (NPTC)
文摘Ascorbate peroxidase, a haem protein (EC 1.11.1.11), efficiently scavenges hydrogen peroxide (H2O2) in cytosol and chloroplasts of plants. In this study, a fulllength coding sequence of thylakoid-bound ascorbate peroxidase cDNA (TatAPX) was cloned from a drought tolerant wheat cultivar C306. Homology modeling of the TatAPX protein was performed by using the template crystal structure of chloroplastic ascorbate peroxidase from tobacco plant (PDB: IIYN). The model structure was further refined by molecular mechanics and dynamic methods using various tools such as PROCHECK, ProSA and Verify3D. The predicted model was then tested for docking with H2O2, the substrate for TatAPX enzyme. The results revealed that Arg233 and Glu255 in the predicted active site of the enzyme are two important amino acid residues responsible for strong hydrogen bonding affinity with H2O2, which might play an important role in scavenging of H2O2 from the plant system.
基金supported by NSFC(Nos50963002 and 51073074)Key Projects in the National Science& Technology Pillar Program in the Eleventh Five-year Plan Period of China(Nos2006BAD02A04 and 2006BAD01A01)Jiangxi Provincial Department of Education(NosGJJ11590 and GJJ10678)
文摘The vitamin C(VC) in crops was successfully determined using ascorbate oxidase(AO) electrochemical biosensor based on the biocompatible poly(3,4-ethylenedioxythiophene)(PEDOT) matrices,which was easily prepared by one-step electrodeposition technique in ionic liquid microemulsions.The fabricated biosensor displayed excellent bioelectrocatalytic performance to the oxidation of VC,wide linear range,low detection limit,fast response time,good operational and storage stability,the good results of the determination of VC in vegetable crops indicated that the fabricated biosensor will be a good candidate for the physiological and biochemical studies of crops in near future.