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Peroxisomal APX knockdown triggers antioxidant mechanisms favourable for coping with high photorespiratory H2O2 induced by CAT deficiency in rice
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RACHEL H. V. SOUSA FABRICIO E. L. CARVALHO CAROL W. RIBEIRO GISELE PASSAIA JULIANA R. CUNHA YUGO LIMA‐MELO MÁRCIA MARGIS‐PINHEIRO JOAQUIM A. G. SILVEIRA 《Plant, cell & environment》2015,38(3):499-513
The physiological role of peroxisomal ascorbate peroxidases (pAPX) is unknown; therefore, we utilized pAPX4 knockdown rice and catalase (CAT) inhibition to assess its role in CAT compensation under high photorespiration. pAPX4 knockdown induced co‐suppression in the expression of pAPX3. The rice mutants exhibited metabolic changes such as lower CAT and glycolate oxidase (GO) activities and reduced glyoxylate content; however, APX activity was not altered. CAT inhibition triggered different changes in the expression of CAT, APX and glutathione peroxidase (GPX) isoforms between non‐transformed (NT) and silenced plants. These responses were associated with alterations in APX, GPX and GO activities, suggesting redox homeostasis differences. The glutathione oxidation‐reduction states were modulated differently in mutants, and the ascorbate redox state was greatly affected in both genotypes. The pAPX suffered less oxidative stress and photosystem II (PSII) damage and displayed higher photosynthesis than the NT plants. The improved acclimation exhibited by the pAPX plants was indicated by lower H2O2 accumulation, which was associated with lower GO activity and glyoxylate content. The suppression of both pAPXs and/or its downstream metabolic and molecular effects may trigger favourable antioxidant and compensatory mechanisms to cope with CAT deficiency. This physiological acclimation may involve signalling by peroxisomal H2O2, which minimized the photorespiration. 相似文献
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以荒漠木本C_3植物天山猪毛菜、C_3-C_4中间型植物松叶猪毛菜、C_4植物木本猪毛菜为研究对象,采用盆栽控水试验,设置正常供水和轻度、中度和重度干旱处理(土壤含水量分别为田间持水量的80%、60%、45%和35%),研究不同程度干旱胁迫对3种不同光合类型荒漠植物叶片超微结构的影响。结果表明:(1)正常水分条件下,叶肉细胞中各细胞器结构完整。(2)轻度干旱胁迫下,3种植物叶片超微结构未受损伤,无明显变化。(3)中度干旱胁迫下,天山猪毛菜和松叶猪毛菜叶肉细胞壁界限不清晰,类囊体片层扩张且排列不紧密,不同之处在于,天山猪毛菜线粒体最先出现降解,内含物流失,而松叶猪毛菜线粒体外膜轮廓变形,嵴减少;木本猪毛菜线粒体无明显变化,叶绿体轻微扩张。(4)重度干旱胁迫下,天山猪毛菜和松叶猪毛菜叶绿体受损且结构混乱,线粒体出现降解;木本猪毛菜叶绿体出现膨胀,线粒体外膜轮廓模糊,嵴减少且结构模糊不清楚。研究认为,不同程度干旱胁迫下木本猪毛菜叶绿体和线粒体的受损程度都最低;干旱胁迫下天山猪毛菜和松叶猪毛菜叶绿体的受损程度大致相似;松叶猪毛菜和木本猪毛菜线粒体对干旱胁迫的耐受力要比叶绿体强。 相似文献
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Jos Manuel Ugalde Philippe Fuchs Thomas Nietzel Edoardo A Cutolo Maria Homagk Ute C Vothknecht Loreto Holuigue Markus Schwarzlnder Stefanie J Müller-Schüssele Andreas J Meyer 《Plant physiology》2021,186(1):125
Metabolic fluctuations in chloroplasts and mitochondria can trigger retrograde signals to modify nuclear gene expression. Mobile signals likely to be involved are reactive oxygen species (ROS), which can operate protein redox switches by oxidation of specific cysteine residues. Redox buffers, such as the highly reduced glutathione pool, serve as reservoirs of reducing power for several ROS-scavenging and ROS-induced damage repair pathways. Formation of glutathione disulfide and a shift of the glutathione redox potential (EGSH) toward less negative values is considered as hallmark of several stress conditions. Here we used the herbicide methyl viologen (MV) to generate ROS locally in chloroplasts of intact Arabidopsis (Arabidopsis thaliana) seedlings and recorded dynamic changes in EGSH and H2O2 levels with the genetically encoded biosensors Grx1-roGFP2 (for EGSH) and roGFP2-Orp1 (for H2O2) targeted to chloroplasts, the cytosol, or mitochondria. Treatment of seedlings with MV caused rapid oxidation in chloroplasts and, subsequently, in the cytosol and mitochondria. MV-induced oxidation was significantly boosted by illumination with actinic light, and largely abolished by inhibitors of photosynthetic electron transport. MV also induced autonomous oxidation in the mitochondrial matrix in an electron transport chain activity-dependent manner that was milder than the oxidation triggered in chloroplasts by the combination of MV and light. In vivo redox biosensing resolves the spatiotemporal dynamics of compartmental responses to local ROS generation and provides a basis for understanding how compartment-specific redox dynamics might operate in retrograde signaling and stress acclimation in plants. Methyl viologen-induced photo-oxidative stress increases hydrogen peroxide and oxidation of glutathione in chloroplasts, cytosol, and mitochondria, as well as autonomous oxidation in mitochondria. 相似文献
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Takumi Ohdate Yoshiharu Inoue 《Biochimica et Biophysica Acta (BBA)/Molecular and Cell Biology of Lipids》2012,1821(9):1295-1305
Saccharomyces cerevisiae is able to use some fatty acids, such as oleic acid, as a sole source of carbon. β-oxidation, which occurs in a single membrane-enveloped organelle or peroxisome, is responsible for the assimilation of fatty acids. In S. cerevisiae, β-oxidation occurs only in peroxisomes, and H2O2 is generated during this fatty acid-metabolizing pathway. S. cerevisiae has three GPX genes (GPX1, GPX2, and GPX3) encoding atypical 2-Cys peroxiredoxins. Here we show that expression of GPX1 was induced in medium containing oleic acid as a carbon source in an Msn2/Msn4-dependent manner. We found that Gpx1 was located in the peroxisomal matrix. The peroxisomal Gpx1 showed peroxidase activity using thioredoxin or glutathione as a reducing power. Peroxisome biogenesis was induced when cells were cultured with oleic acid. Peroxisome biogenesis was impaired in gpx1? cells, and subsequently, the growth of gpx1? cells was lowered in oleic acid-containing medium. Gpx1 contains six cysteine residues. Of the cysteine-substituted mutants of Gpx1, Gpx1C36S was not able to restore growth and peroxisome formation in oleic acid-containing medium, therefore, redox regulation of Gpx1 seems to be involved in the mechanism of peroxisome formation. 相似文献
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植物谷胱甘肽过氧化物酶(glutathione peroxidase,GPX)是清除体内活性氧的一种关键酶,在植物抗逆反应中发挥重要作用.本研究从水稻中克隆到2个GPX基因,分别为OsGPX3和OsGPX4.OsGPX3和OsGPX4分别编码238和234个氨基酸组成的蛋白质,预测分子量分别是25.84 kD和25.07 kD.两个基因都包含5个内含子,但是两个基因所对应的内含子长度具有较大变异.组织表达谱分析发现这2个基因在根、茎、叶和叶鞘中均表达,是组成型表达基因.在大肠杆菌中表达并纯化了这2个基因的重组蛋白,酶活性分析显示OsGPX3和OsGPX4蛋白对底物H2O2、tBOOH和COOH具有较高活性,但是OsGPX3对3种底物的活性均高于OsGPX4,蛋白质酶活性的差异预示着这2个基因可能存在功能上的分化. 相似文献
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Yugo Lima‐Melo Fabricio E. L. Carvalho Márcio O. Martins Gisele Passaia Rachel H. V. Sousa Milton C. Lima Neto Márcia Margis‐Pinheiro Joaquim A. G. Silveira 《植物学报(英文版)》2016,58(8):737-748
The physiological role of plant mitochondrial glutathione peroxidases is scarcely known. This study attempted to elucidate the role of a rice mitochondrial isoform(GPX1) in photosynthesis under normal growth and salinity conditions. GPX1 knockdown rice lines(GPX1s) were tested in absence and presence of 100 mM NaCl for 6 d.Growth reduction of GPX1 s line under non-stressful conditions, compared with non-transformed(NT) plants occurred in parallel to increased H_2O_2 and decreased GSH contents. These changes occurred concurrently with photosynthesis impairment, particularly in Calvin cycle's reactions, since photochemical efficiency did not change.Thus, GPX1 silencing and downstream molecular/metabolic changes modulated photosynthesis differentially. In contrast, salinity induced reduction in both phases of photosynthesis, which were more impaired in silenced plants.These changes were associated with root morphology alterations but not shoot growth. Both studied lines displayed increased GPX activity but H_2O_2 content did not change in response to salinity. Transformed plants exhibited lower photorespiration, water use efficiency and root growth, indicating that GPX1 could be important to salt tolerance. Growth reduction of GPX1 s line might be related to photosynthesis impairment, which in turn could have involved a cross talk mechanism between mitochondria and chloroplast originated from redox changes due to GPX1 deficiency. 相似文献
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Aberrant chloroplasts in transgenic rice plants expressing a high level of maize NADP-dependent malic enzyme 总被引:16,自引:0,他引:16
NADP-dependent malic enzyme (NADP-ME) is a major decarboxylating enzyme in NADP-ME-type C4 species such as maize and Flaveria. In this study, chloroplastic NADP-ME was transferred to rice (Oryza sativa L.) using a chimeric gene composed of maize NADP-ME cDNA under the control of rice light-harvesting chlorophyll-a/b-binding protein (Cab) promoter. There was a 20- to 70-fold increase in the NADP-ME activity in leaves of transgenic rice
compared to that in wild-type rice plants. Immunocytochemical studies by electron microscopy showed that maize NADP-ME was
mostly localized in chloroplasts in transgenic rice plants, and that the chloroplasts were agranal without thylakoid stacking.
Chlorophyll content and photosystem II activity were inversely correlated with the level of NADP-ME activity. These results
suggest that aberrant chloroplasts in transgenic plants may be caused by excessive NADP-ME activity. Based on these results
and the known fact that only bundle sheath cells of NADP-ME species, among all three C4 subgroups, have agranal chloroplasts, we postulate that a high level of chloroplastic NADP-ME activity could strongly affect
the development of chloroplasts.
Received: 27 January 1999 / Accepted: 20 January 2000 相似文献
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Yiwei Du Wei He Changwang Deng Xi Chen Lanming Gou Fugui Zhu Wei Guo Jianfu Zhang Tao Wang 《PloS one》2016,11(3)
Flowering time is a critical trait for crops cultivated under various temperature/photoperiod conditions around the world. To understand better the flowering time of rice, we used the vector pTCK303 to produce several lines of RNAi knockdown transgenic rice and investigated their flowering times and other agronomic traits. Among them, the heading date of FRRP1-RNAi knockdown transgenic rice was 23–26 days earlier than that of wild-type plants. FRRP1 is a novel rice gene that encodes a C3HC4-type Really Interesting Novel Gene (RING) finger domain protein. In addition to the early flowering time, FRRP1-RNAi knockdown transgenic rice caused changes on an array of agronomic traits, including plant height, panicle length and grain length. We analyzed the expression of some key genes associated with the flowering time and other agronomic traits in the FRRP1-RNAi knockdown lines and compared with that in wild-type lines. The expression of Hd3a increased significantly, which was the key factor in the early flowering time. Further experiments showed that the level of histone H2B monoubiquitination (H2Bub1) was noticeably reduced in the FRRP1-RNAi knockdown transgenic rice lines compared with wild-type plants and MBP-FRRP1-F1 was capable of self-ubiquitination. The results indicate that Flowering Related RING Protein 1 (FRRP1) is involved in histone H2B monoubiquitination and suggest that FRRP1 functions as an E3 ligase in vivo and in vitro. In conclusion, FRRP1 probably regulates flowering time and yield potential in rice by affecting histone H2B monoubiquitination, which leads to changes in gene expression in multiple processes. 相似文献
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Molecular and Catalytic Properties of Glutathione Peroxidase Family Proteins from Pinus tabulaeformis 总被引:1,自引:0,他引:1
Glutathione peroxidase (GPX) is one of the key enzymes that protect cells against oxidative damage caused by reactive oxygen species. Previous studies of plant GPXs focused mainly on angiosperms. In contrast, little information is available on the molecular characteristics of this gene family in gymnosperms. In this study, four GPX genes (PtaGPX1, 2, 3, and 4) were cloned from the gymnosperm Pinus tabulaeformis, which showed high protein sequence identity and similar expression patterns in various tissues. The four Pinus GPX proteins were expressed in Escherichia coli, and the purified proteins used thioredoxin, but not glutathione, as an electron donor. The four Pinus GPXs showed different enzymatic activities and kinetic characteristics, suggesting functional divergence. Two conserved Cys residues (corresponding to Cys44 and Cys92 of PtaGPX3) were identified in all plant GPXs, and their functions were assessed using site-directed mutagenesis. Cys44 and Cys92 of PtaGPX3 could form an intramolecular disulfide bond under oxidizing conditions. These two residues were critical components of active sites and contributed to catalytic activity. This study provides novel insights into the functional divergence and catalytic properties of the GPX family in gymnosperms. 相似文献
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Photosynthesis of Grass Species Differing in Carbon Dioxide Fixation Pathways : VIII. Ultrastructural Characteristics of Panicum Species in the Laxa Group 总被引:2,自引:2,他引:0
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Ultrastructural studies of leaves of seven Panicum species in or closely related to the Laxa group and classified as C3, C4 or C3-C4 intermediate were undertaken to examine features associated with C3 and C4 photosynthesis. The C3 species Panicum rivulare Trin. had few organelles in bundle sheath cell profiles (2 chloroplasts, 1.1 mitochondria, and 0.3 peroxisomes per cell section) compared to an average of 10.6 chloroplasts, 17.7 mitochondria, and 3.2 peroxisomes per bundle sheath cell profile for three C3-C4 species, Panicum milioides Nees ex Trin., Panicum decipiens Nees ex Trin. and Panicum schenckii Hack. However, two other C3 species, Panicum laxum Sw. and Panicum hylaeicum Mez, contained about 0.7, 0.5, and 0.3 as many chloroplasts, mitochondria, and peroxisomes, respectively, as in bundle sheath cell profiles of the C3-C4 species. Chloroplasts and mitochondria in bundle sheath cells were larger than those in mesophyll cells for the C4 species Panicum prionitis Griseb. and the C3-C4 species, but in C3 species the organelles were similar in size or were smaller in the bundle sheath cells. The C3-C4 species and P. laxum and P. hylaeicum exhibited an unusually close association of organelles in bundle sheath cells with mitochondria frequently surrounded in profile by chloroplasts. The high concentrations in bundle sheath cells of somewhat larger organelles than in mesophyll cells correlates with the reduced photorespiration of the C3-C4 species. 相似文献
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Dipali Rani Gupta Sanjida Khanom Md. Motiar Rohman Mirza Hasanuzzaman Musrat Zahan Surovy Nur Uddin Mahmud Md. Robyul Islam Ashifur Rahman Shawon Mahfuzur Rahman Kamel A. Abd-Elsalam Tofazzal Islam 《Physiology and Molecular Biology of Plants》2021,27(9):2127
Wheat blast caused by the hemibiotroph fungal pathogen Magnaporthe oryzae Triticum (MoT) pathotype is a destructive disease of wheat in South America, Bangladesh and Zambia. This study aimed to determine and compare the activities of antioxidant enzymes in susceptible (wheat, maize, barley and swamp rice grass) and resistant (rice) plants when interacting with MoT. The activities of reactive oxygen species-detoxifying enzymes; catalase (CAT), ascorbate peroxidase (APX), glutathione peroxidase (GPX), glutathione S-transferase (GST), peroxidase (POX) were increased in all plants in response to MoT inoculation with a few exceptions. Interestingly, an early and very high activity of CAT was observed within 24 h after inoculation in wheat, barley, maize and swamp rice grass with lower H2O2 concentration. In contrast, an early and high accumulation of H2O2 was observed in rice at 48 hai with little CAT activity only at a later stage of MoT inoculation. The activities of APX, GST and POD were also high at an early stage of infection in rice. However, these enzymes activities were very high at a later stage in wheat, barley, maize and swamp rice grass. The activity of GPX gradually decreased with the increase of time in rice. Taken together, our results suggest that late and early inductions of most of the antioxidant enzyme activities occurs in susceptible and resistant plants, respectively. This study demonstrates some insights into physiological responses of host and non-host plants when interacting with the devastating wheat blast fungus MoT, which could be useful for developing blast resistant wheat. 相似文献
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In Panicum species of the Laxa group, some of which have characteristics intermediate to C3 and C4 photosynthesis species, some mitochondria in leaf bundle sheath cells are surrounded by chloroplasts when viewed in profile. Serial sectioning of leaves of one Laxa species, Panicum schenckii Hack, shows that these mitochondria are enclosed by chloroplasts. Complete enclosure rather than invagination also is indicated by absence of two concentric chloroplast membranes surrounding the mitochondrial profiles. 相似文献
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S. Y. Shin Y. S. Kim I. S. Kim Y. H. Kim H. M. Park H. S. Yoon 《Biologia Plantarum》2014,58(3):456-468
In chloroplasts and mitochondria, antioxidant mechanisms include the ascorbate-glutathione cycle, and monodehydroascorbate reductase (MDHAR) is important for regeneration of ascorbate (AsA) from monodehydroascorbate (MDHA). To improve detoxification of reactive oxygen species (ROS), we established a construct of the MDHAR gene from Brassica rapa fused to the targeting signal peptides of Pisum sativum glutathione reductase (GR), which was controlled by a stress-inducible SWPA2 promoter, and introduced this expression system into Arabidopsis thaliana. Transgenic (TG) plants overexpressing BrMDHAR targeted to chloroplasts and mitochondria through signal peptides showed an elevated MDHAR activity and an increased ratio of AsA to dehydroascorbate (DHA) when compared to wild-type (WT) plants under a freezing stress. These led to increased photosynthetic parameters, redox homeostasis, and biomass in TG plants when compared to the WT plants. Our results suggest that the overexpression of the BrMDHAR gene targeted to chloroplasts and mitochondria conferred an enhanced tolerance against the freezing stress, and a stress adaptation of dual-targeted BrMDHAR was better than that of single BrMDHAR. 相似文献
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Sílvia B. Rosa Andréia Caverzan Felipe K. Teixeira Fernanda Lazzarotto Joaquim A.G. Silveira Sérgio Luiz Ferreira-Silva João Abreu-Neto Rogério Margis Márcia Margis-Pinheiro 《Phytochemistry》2010,71(5-6):548-558
Ascorbate peroxidases (APX, EC 1.1.11.1) are class I heme-peroxidases, which catalyze the conversion of H2O2 into H2O, using ascorbate as a specific electron donor. Previously, the presence of eight Apx genes was identified in the nuclear genome of rice (Oryza sativa), encoding isoforms that are located in different sub-cellular compartments. Herein, the generation of rice transgenic plants silenced for either both or each one of the cytosolic Apx1 and Apx2 genes was carried out in order to investigate the importance of cytosolic Apx isoforms on plant development and on plant stress responses. Transgenic double Apx1/2-silenced plants exhibited normal development, even though these plants showed a global reduction of Apx activity which strongly impacts the whole antioxidant system regulation. Apx1/2-silenced plants also showed increased H2O2 accumulation under control and stress situations and presented higher tolerance to toxic concentration of aluminum when compared to wild type plants. On the other hand, silencing OsApx1 and OsApx2 genes individually resulted in strong effect on plant development producing semi-dwarf phenotype. These results suggested that the double silencing of cytosolic OsApx genes induced compensatory antioxidant mechanisms in rice while single knockdown of these genes did not, which resulted in the impairing of normal plant development. 相似文献
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Protoplasm from Bryopsis maxima, a coenocytic green alga, was dissociated into two fractions; chloroplasts and a protoplasmic fraction without chloroplasts (PF). PF induced nuclei, mitochondria, dictyosome, endplasmic reticulum, etc. These two fractions were reunited and protoplasts with complete plasmamembrane were reformed within 10 h. Cell wall regeneration was observed 24–30 h after reuniting. New cells began budding 2–3 days after, and 1 month after reunification, they had developed into mature plants. 相似文献
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Glutamine Synthetase/Glutamine: alpha-Ketoglutarate Aminotransferase in Chloroplasts from the Marine Alga Caulerpa simpliciuscula
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The enzymic capacities for ammonia assimilation into amino acids have been investigated in chloroplasts from the siphonous green alga Caulerpa simpliciuscula (Turner) C. Ag. The results show that these chloroplasts differ from those of higher plants in having present simultaneously the enzymic capacities to permit assimilation of ammonia by two pathways. Glutamine synthetase (EC 6.3.1.2) activity at levels up to 4 μmoles per mg chlorophyll per hour were found in soluble extracts of the chloroplasts. Glutamine(amide):α-ketoglutarate aminotransferase (oxidoreductase ferredoxin) (EC 1.4.7.1) activity at levels up to 1.4 μmoles per mg chlorophyll per hour was detected by incubation of photosynthetically active chloroplasts either in light or with reduced ferredoxin. Together these enzymes provide the capacity for the conventional pathway of ammonium assimilation in chloroplasts via glutamine. A similar level of a glutamate dehydrogenase with an unusually low Km for ammonia which has been described previously in these chloroplasts provides the second potential pathway. 相似文献