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1.
Plant J ; 114(4): 836-854, 2023 05.
Article in English | MEDLINE | ID: mdl-36883867

ABSTRACT

Arabidopsis histone deacetylase HDA19 is required for gene expression programs of a large spectrum of plant developmental and stress-responsive pathways. How this enzyme senses cellular environment to control its activity remains unclear. In this work, we show that HDA19 is post-translationally modified by S-nitrosylation at 4 Cysteine (Cys) residues. HDA19 S-nitrosylation depends on the cellular nitric oxide level, which is enhanced under oxidative stress. We find that HDA19 is required for cellular redox homeostasis and plant tolerance to oxidative stress, which in turn stimulates its nuclear enrichment, S-nitrosylation and epigenetic functions including binding to genomic targets, histone deacetylation and gene repression. The Cys137 of the protein is involved in basal and stress-induced S-nitrosylation, and is required for HDA19 functions in developmental, stress-responsive and epigenetic controls. Together, these results indicate that S-nitrosylation regulates HDA19 activity and is a mechanism of redox-sensing for chromatin regulation of plant tolerance to stress.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Histone Deacetylases/genetics , Histone Deacetylases/metabolism , Chromatin/metabolism , Nitric Oxide/metabolism
2.
J Exp Bot ; 75(15): 4549-4572, 2024 Aug 12.
Article in English | MEDLINE | ID: mdl-38676714

ABSTRACT

Redox reactions are fundamental to energy conversion in living cells, and also determine and tune responses to the environment. Within this context, the tripeptide glutathione plays numerous roles. As an important antioxidant, glutathione confers redox stability on the cell and also acts as an interface between signalling pathways and metabolic reactions that fuel growth and development. It also contributes to the assembly of cell components, biosynthesis of sulfur-containing metabolites, inactivation of potentially deleterious compounds, and control of hormonal signalling intensity. The multiplicity of these roles probably explains why glutathione status has been implicated in influencing plant responses to many different conditions. In particular, there is now a considerable body of evidence showing that glutathione is a crucial player in governing the outcome of biotic stresses. This review provides an overview of glutathione synthesis, transport, degradation, and redox turnover in plants. It examines the expression of genes associated with these processes during pathogen challenge and related conditions, and considers the diversity of mechanisms by which glutathione can influence protein function and gene expression.


Subject(s)
Glutathione , Oxidation-Reduction , Plants , Glutathione/metabolism , Plants/metabolism , Signal Transduction , Gene Expression Regulation, Plant
3.
Plant Physiol ; 182(2): 1161-1181, 2020 02.
Article in English | MEDLINE | ID: mdl-31659127

ABSTRACT

Plants optimize their growth and survival through highly integrated regulatory networks that coordinate defensive measures and developmental transitions in response to environmental cues. Protein phosphatase 2A (PP2A) is a key signaling component that controls stress reactions and growth at different stages of plant development, and the PP2A regulatory subunit PP2A-B'γ is required for negative regulation of pathogenesis responses and for maintenance of cell homeostasis in short-day conditions. Here, we report molecular mechanisms by which PP2A-B'γ regulates Botrytis cinerea resistance and leaf senescence in Arabidopsis (Arabidopsis thaliana). We extend the molecular functionality of PP2A-B'γ to a protein kinase-phosphatase interaction with the defense-associated calcium-dependent protein kinase CPK1 and present indications this interaction may function to control CPK1 activity. In presenescent leaf tissues, PP2A-B'γ is also required to negatively control the expression of salicylic acid-related defense genes, which have recently proven vital in plant resistance to necrotrophic fungal pathogens. In addition, we find the premature leaf yellowing of pp2a-b'γ depends on salicylic acid biosynthesis via SALICYLIC ACID INDUCTION DEFICIENT2 and bears the hallmarks of developmental leaf senescence. We propose PP2A-B'γ age-dependently controls salicylic acid-related signaling in plant immunity and developmental leaf senescence.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/enzymology , Botrytis/immunology , Cellular Senescence/genetics , Disease Resistance/genetics , Plant Diseases/immunology , Plant Leaves/metabolism , Protein Phosphatase 2/metabolism , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis/microbiology , Arabidopsis Proteins/genetics , Calcium/metabolism , Cellular Senescence/physiology , Cysteine Endopeptidases/genetics , Cysteine Endopeptidases/metabolism , Disease Resistance/immunology , Gene Expression Regulation, Developmental/genetics , Gene Expression Regulation, Plant/genetics , Genotype , Intramolecular Transferases/genetics , Intramolecular Transferases/metabolism , Mutation , Phenotype , Plant Diseases/genetics , Plant Diseases/microbiology , Plant Immunity/genetics , Plant Leaves/genetics , Plant Leaves/growth & development , Protein Binding , Protein Kinases/genetics , Protein Kinases/metabolism , Protein Phosphatase 2/genetics , Salicylic Acid/metabolism , Signal Transduction/genetics , Transcription Factors/genetics , Transcription Factors/metabolism , Transcriptome/genetics
4.
Semin Cell Dev Biol ; 80: 3-12, 2018 08.
Article in English | MEDLINE | ID: mdl-28733165

ABSTRACT

As sessile oxygenic organisms with a plastic developmental programme, plants are uniquely positioned to exploit reactive oxygen species (ROS) as powerful signals. Plants harbor numerous ROS-generating pathways, and these oxidants and related redox-active compounds have become tightly embedded into plant function and development during the course of evolution. One dominant view of ROS-removing systems sees them as beneficial antioxidants battling to keep damaging ROS below dangerous levels. However, it is now established that ROS are a necessary part of subcellular and intercellular communication in plants and that some of their signaling functions require ROS-metabolizing systems. For these reasons, it is suggested that "ROS processing systems" would be a more accurate term than "antioxidative systems" to describe cellular components that are most likely to interact with ROS and, in doing so, transmit oxidative signals. Within this framework, our update provides an overview of the complexity and compartmentation of ROS production and removal. We place particular emphasis on the importance of ROS-interacting systems such as the complex cellular thiol network in the redox regulation of phytohormone signaling pathways that are crucial for plant development and defense against external threats.


Subject(s)
Antioxidants/pharmacology , Oxidation-Reduction/drug effects , Oxygen/metabolism , Plant Physiological Phenomena/drug effects , Reactive Oxygen Species/metabolism , Animals , Humans , Oxidative Stress/physiology
5.
Plant Cell Environ ; 43(5): 1175-1191, 2020 05.
Article in English | MEDLINE | ID: mdl-31990075

ABSTRACT

Photorespiratory hydrogen peroxide (H2 O2 ) plays key roles in pathogenesis responses by triggering the salicylic acid (SA) pathway in Arabidopsis. However, factors linking intracellular H2 O2 to activation of the SA pathway remain elusive. In this work, the catalase-deficient Arabidopsis mutant, cat2, was exploited to elucidate the impact of S-nitrosoglutathione reductase 1 (GSNOR1) on H2 O2 -dependent signalling pathways. Introducing the gsnor1-3 mutation into the cat2 background increased S-nitrosothiol levels and abolished cat2-triggered cell death, SA accumulation, and associated gene expression but had little additional effect on the major components of the ascorbate-glutathione system or glycolate oxidase activities. Differential transcriptome profiles between gsnor1-3 and cat2 gsnor1-3 together with damped ROS-triggered gene expression in cat2 gsnor1-3 further indicated that GSNOR1 acts to mediate the SA pathway downstream of H2 O2 . Up-regulation of GSNOR activity was compromised in cat2 cad2 and cat2 pad2 mutants in which glutathione accumulation was genetically prevented. Experiments with purified recombinant GSNOR revealed that the enzyme is posttranslationally regulated by direct denitrosation in a glutathione-dependent manner. Together, our findings identify GSNOR1-controlled nitrosation as a key factor in activation of the SA pathway by H2 O2 and reveal that glutathione is required to maintain this biological function.


Subject(s)
Arabidopsis Proteins/metabolism , Glutathione Reductase/metabolism , Glutathione/metabolism , Hydrogen Peroxide/metabolism , Oxidation-Reduction , Signal Transduction , Arabidopsis/metabolism , Gene Expression Regulation, Plant , Microscopy, Confocal , Nitrosation , Oxidative Stress , Plant Growth Regulators/metabolism , Real-Time Polymerase Chain Reaction , Salicylic Acid/metabolism
6.
Plant Cell Environ ; 42(2): 688-700, 2019 02.
Article in English | MEDLINE | ID: mdl-30291629

ABSTRACT

Three genes encode catalase in Arabidopsis. Although the role of CAT2 in photorespiration is well established, the importance of the different catalases in other processes is less clear. Analysis of cat1, cat2, cat3, cat1 cat2, and cat2 cat3 T-DNA mutants revealed that cat2 had the largest effect on activity in both roots and leaves. Root growth was inhibited in all cat2-containing lines, but this inhibition was prevented by growing plants at high CO2 , suggesting that it is mainly an indirect effect of stress at the leaf level. Analysis of double mutants suggested some overlap between CAT2 and CAT3 functions in leaves and CAT1 and CAT2 in seeds. When plants had been grown to a similar developmental stage in short days or long days, equal-time exposure to oxidative stress caused by genetic or pharmacological inhibition of catalase produced a much stronger induction of H2 O2 marker genes in short day plants. Together, our data (a) underline the importance of CAT2 in basal H2 O2 processing in Arabidopsis; (b) suggest that CAT1 and CAT3 are mainly "backup" or stress-specific enzymes; and (c) establish that day length-dependent responses to catalase deficiency are independent of the duration of oxidative stress.


Subject(s)
Arabidopsis Proteins/physiology , Arabidopsis/growth & development , Oxidation-Reduction , Arabidopsis/embryology , Arabidopsis/enzymology , Arabidopsis/physiology , Arabidopsis Proteins/metabolism , Catalase/metabolism , Catalase/physiology , Photoperiod , Plant Leaves/enzymology , Plant Leaves/growth & development , Plant Roots/enzymology , Plant Roots/growth & development , Reactive Oxygen Species/metabolism , Signal Transduction
8.
Plant Cell ; 28(8): 1844-59, 2016 08.
Article in English | MEDLINE | ID: mdl-27432873

ABSTRACT

Hydrogen peroxide (H2O2) can act as a signaling molecule that influences various aspects of plant growth and development, including stress signaling and cell death. To analyze molecular mechanisms that regulate the response to increased H2O2 levels in plant cells, we focused on the photorespiration-dependent peroxisomal H2O2 production in Arabidopsis thaliana mutants lacking CATALASE2 (CAT2) activity (cat2-2). By screening for second-site mutations that attenuate the PSII maximum efficiency (Fv'/Fm') decrease and lesion formation linked to the cat2-2 phenotype, we discovered that a mutation in SHORT-ROOT (SHR) rescued the cell death phenotype of cat2-2 plants under photorespiration-promoting conditions. SHR deficiency attenuated H2O2-dependent gene expression, oxidation of the glutathione pool, and ascorbate depletion in a cat2-2 genetic background upon exposure to photorespiratory stress. Decreased glycolate oxidase and catalase activities together with accumulation of glycolate further implied that SHR deficiency impacts the cellular redox homeostasis by limiting peroxisomal H2O2 production. The photorespiratory phenotype of cat2-2 mutants did not depend on the SHR functional interactor SCARECROW and the sugar signaling component ABSCISIC ACID INSENSITIVE4, despite the requirement for exogenous sucrose for cell death attenuation in cat2-2 shr-6 double mutants. Our findings reveal a link between SHR and photorespiratory H2O2 production that has implications for the integration of developmental and stress responses.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Catalase/metabolism , Transcription Factors/deficiency , Transcription Factors/metabolism , Arabidopsis/cytology , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Catalase/genetics , Cell Death/genetics , Cell Death/physiology , Gene Expression Regulation, Plant/genetics , Gene Expression Regulation, Plant/physiology , Hydrogen Peroxide/metabolism , Oxidation-Reduction , Plants, Genetically Modified/cytology , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Signal Transduction/genetics , Signal Transduction/physiology , Transcription Factors/genetics
9.
Plant Physiol ; 174(2): 956-971, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28381499

ABSTRACT

The complexity of plant antioxidative systems gives rise to many unresolved questions. One relates to the functional importance of dehydroascorbate reductases (DHARs) in interactions between ascorbate and glutathione. To investigate this issue, we produced a complete set of loss-of-function mutants for the three annotated Arabidopsis (Arabidopsis thaliana) DHARs. The combined loss of DHAR1 and DHAR3 expression decreased extractable activity to very low levels but had little effect on phenotype or ascorbate and glutathione pools in standard conditions. An analysis of the subcellular localization of the DHARs in Arabidopsis lines stably transformed with GFP fusion proteins revealed that DHAR1 and DHAR2 are cytosolic while DHAR3 is chloroplastic, with no evidence for peroxisomal or mitochondrial localizations. When the mutations were introduced into an oxidative stress genetic background (cat2), the dhar1 dhar2 combination decreased glutathione oxidation and inhibited cat2-triggered induction of the salicylic acid pathway. These effects were reversed in cat2 dhar1 dhar2 dhar3 complemented with any of the three DHARs. The data suggest that (1) DHAR can be decreased to negligible levels without marked effects on ascorbate pools, (2) the cytosolic isoforms are particularly important in coupling intracellular hydrogen peroxide metabolism to glutathione oxidation, and (3) DHAR-dependent glutathione oxidation influences redox-driven salicylic acid accumulation.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/enzymology , Chloroplasts/enzymology , Cytosol/enzymology , Oxidative Stress , Oxidoreductases/metabolism , Salicylic Acid/metabolism , Antioxidants/metabolism , Arabidopsis/metabolism , Ascorbic Acid/metabolism , Cell Death , DNA, Bacterial/genetics , Genetic Complementation Test , Glutathione/metabolism , Green Fluorescent Proteins/metabolism , Mutagenesis, Insertional/genetics , Mutation/genetics , Phenotype , Recombinant Fusion Proteins/metabolism , Subcellular Fractions/metabolism
10.
Biochem J ; 474(6): 877-883, 2017 03 07.
Article in English | MEDLINE | ID: mdl-28270560

ABSTRACT

Concepts of the roles of reactive oxygen species (ROS) in plants and animals have shifted in recent years from focusing on oxidative damage effects to the current view of ROS as universal signalling metabolites. Rather than having two opposing activities, i.e. damage and signalling, the emerging concept is that all types of oxidative modification/damage are involved in signalling, not least in the induction of repair processes. Examining the multifaceted roles of ROS as crucial cellular signals, we highlight as an example the loss of photosystem II function called photoinhibition, where photoprotection has classically been conflated with oxidative damage.


Subject(s)
Light Signal Transduction/physiology , Photosynthesis/physiology , Photosystem II Protein Complex/physiology , Plants/radiation effects , Reactive Oxygen Species/metabolism , Adenosine Triphosphate/biosynthesis , Chlorophyll/agonists , Chlorophyll/metabolism , Chloroplasts/metabolism , Chloroplasts/radiation effects , Chloroplasts/ultrastructure , Fluorescence , Light , NADP/biosynthesis , Oxidation-Reduction , Oxidative Stress , Plants/metabolism , Plants/ultrastructure
11.
Plant Physiol ; 172(2): 929-942, 2016 10.
Article in English | MEDLINE | ID: mdl-27578552

ABSTRACT

Industrial activities have caused tropospheric CO2 concentrations to increase over the last two centuries, a trend that is predicted to continue for at least the next several decades. Here, we report that growth of plants in a CO2-enriched environment activates responses that are central to defense against pathogenic attack. Salicylic acid accumulation was triggered by high-growth CO2 in Arabidopsis (Arabidopsis thaliana) and other plants such as bean (Phaseolus vulgaris). A detailed analysis in Arabidopsis revealed that elevated CO2 primes multiple defense pathways, leading to increased resistance to bacterial and fungal challenge. Analysis of gene-specific mutants provided no evidence that activation of plant defense pathways by high CO2 was caused by stomatal closure. Rather, the activation is partly linked to metabolic effects involving redox signaling. In support of this, genetic modification of redox components (glutathione contents and NADPH-generating enzymes) prevents full priming of the salicylic acid pathway and associated resistance by high CO2 The data point to a particularly influential role for the nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase, a cytosolic enzyme whose role in plants remains unclear. Our observations add new information on relationships between high CO2 and oxidative signaling and provide novel insight into plant stress responses in conditions of increased CO2.


Subject(s)
Arabidopsis/metabolism , Carbon Dioxide/metabolism , Phaseolus/metabolism , Salicylic Acid/metabolism , Arabidopsis/genetics , Arabidopsis/microbiology , Botrytis/physiology , Disease Resistance/genetics , Gene Expression Regulation, Plant , Glutathione/metabolism , Glyceraldehyde-3-Phosphate Dehydrogenases/genetics , Glyceraldehyde-3-Phosphate Dehydrogenases/metabolism , Host-Pathogen Interactions , Mutation , NADP/metabolism , Oxidation-Reduction , Phaseolus/genetics , Phaseolus/microbiology , Plant Diseases/genetics , Plant Diseases/microbiology , Plant Proteins/genetics , Plant Proteins/metabolism , Pseudomonas syringae/physiology , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction/genetics
12.
Plant Physiol ; 171(3): 1581-92, 2016 07.
Article in English | MEDLINE | ID: mdl-27208308

ABSTRACT

Recent years have witnessed enormous progress in understanding redox signaling related to reactive oxygen species (ROS) in plants. The consensus view is that such signaling is intrinsic to many developmental processes and responses to the environment. ROS-related redox signaling is tightly wedded to compartmentation. Because membranes function as barriers, highly redox-active powerhouses such as chloroplasts, peroxisomes, and mitochondria may elicit specific signaling responses. However, transporter functions allow membranes also to act as bridges between compartments, and so regulated capacity to transmit redox changes across membranes influences the outcome of triggers produced at different locations. As well as ROS and other oxidizing species, antioxidants are key players that determine the extent of ROS accumulation at different sites and that may themselves act as signal transmitters. Like ROS, antioxidants can be transported across membranes. In addition, the intracellular distribution of antioxidative enzymes may be modulated to regulate or facilitate redox signaling appropriate to the conditions. Finally, there is substantial plasticity in organellar shape, with extensions such as stromules, peroxules, and matrixules playing potentially crucial roles in organelle-organelle communication. We provide an overview of the advances in subcellular compartmentation, identifying the gaps in our knowledge and discussing future developments in the area.


Subject(s)
Cytosol/metabolism , Plant Cells/metabolism , Reactive Oxygen Species/metabolism , Antioxidants/metabolism , Biological Transport , Cell Compartmentation , Cell Nucleus/metabolism , Hydrogen Peroxide/metabolism , Oxidation-Reduction , Plants/metabolism , Signal Transduction
14.
Plant Physiol ; 171(3): 1720-33, 2016 07.
Article in English | MEDLINE | ID: mdl-27246095

ABSTRACT

In the last decade, microarray studies have delivered extensive inventories of transcriptome-wide changes in messenger RNA levels provoked by various types of oxidative stress in Arabidopsis (Arabidopsis thaliana). Previous cross-study comparisons indicated how different types of reactive oxygen species (ROS) and their subcellular accumulation sites are able to reshape the transcriptome in specific manners. However, these analyses often employed simplistic statistical frameworks that are not compatible with large-scale analyses. Here, we reanalyzed a total of 79 Affymetrix ATH1 microarray studies of redox homeostasis perturbation experiments. To create hierarchy in such a high number of transcriptomic data sets, all transcriptional profiles were clustered on the overlap extent of their differentially expressed transcripts. Subsequently, meta-analysis determined a single magnitude of differential expression across studies and identified common transcriptional footprints per cluster. The resulting transcriptional footprints revealed the regulation of various metabolic pathways and gene families. The RESPIRATORY BURST OXIDASE HOMOLOG F-mediated respiratory burst had a major impact and was a converging point among several studies. Conversely, the timing of the oxidative stress response was a determining factor in shaping different transcriptome footprints. Our study emphasizes the need to interpret transcriptomic data sets in a systematic context, where initial, specific stress triggers can converge to common, aspecific transcriptional changes. We believe that these refined transcriptional footprints provide a valuable resource for assessing the involvement of ROS in biological processes in plants.


Subject(s)
Arabidopsis/genetics , Gene Expression Profiling/methods , Gene Expression Regulation, Plant , Oxidative Stress/genetics , Reactive Oxygen Species/metabolism , Arabidopsis/physiology , Arabidopsis Proteins/genetics , Models, Biological , Multigene Family , Oligonucleotide Array Sequence Analysis , Oxidation-Reduction , Transcription, Genetic
15.
Plant Cell Environ ; 39(5): 951-64, 2016 May.
Article in English | MEDLINE | ID: mdl-26264148

ABSTRACT

Reactive oxygen species (ROS) have a profound influence on almost every aspect of plant biology. Here, we emphasize the fundamental, intimate relationships between light-driven reductant formation, ROS, and oxidative stress, together with compartment-specific differences in redox buffering and the perspectives for their analysis. Calculations of approximate H2 O2 concentrations in the peroxisomes are provided, and based on the likely values in other locations such as chloroplasts, we conclude that much of the H2 O2 detected in conventional in vitro assays is likely to be extracellular. Within the context of scant information on ROS perception mechanisms, we consider current knowledge, including possible parallels with emerging information on oxygen sensing. Although ROS can sometimes be signals for cell death, we consider that an equally important role is to transmit information from metabolism to allow appropriate cellular responses to developmental and environmental changes. Our discussion speculates on novel sensing mechanisms by which this could happen and how ROS could be counted by the cell, possibly as a means of monitoring metabolic flux. Throughout, we place emphasis on the positive effects of ROS, predicting that in the coming decades they will increasingly be defined as hallmarks of viability within a changing and challenging environment.


Subject(s)
Reactive Oxygen Species/metabolism , Signal Transduction , Stress, Physiological , Acclimatization , Homeostasis , Oxidation-Reduction
16.
Plant Cell Environ ; 39(5): 1140-60, 2016 May.
Article in English | MEDLINE | ID: mdl-26864619

ABSTRACT

Oxidative stress and reactive oxygen species (ROS) are common to many fundamental responses of plants. Enormous and ever-growing interest has focused on this research area, leading to an extensive literature that documents the tremendous progress made in recent years. As in other areas of plant biology, advances have been greatly facilitated by developments in genomics-dependent technologies and the application of interdisciplinary techniques that generate information at multiple levels. At the same time, advances in understanding ROS are fundamentally reliant on the use of biochemical and cell biology techniques that are specific to the study of oxidative stress. It is therefore timely to revisit these approaches with the aim of providing a guide to convenient methods and assisting interested researchers in avoiding potential pitfalls. Our critical overview of currently popular methodologies includes a detailed discussion of approaches used to generate oxidative stress, measurements of ROS themselves, determination of major antioxidant metabolites, assays of antioxidative enzymes and marker transcripts for oxidative stress. We consider the applicability of metabolomics, proteomics and transcriptomics approaches and discuss markers such as damage to DNA and RNA. Our discussion of current methodologies is firmly anchored to future technological developments within this popular research field.


Subject(s)
Antioxidants/metabolism , Data Collection , Oxidative Stress , Base Sequence , Biomarkers/metabolism , Metabolomics , Reactive Oxygen Species/metabolism
17.
Plant Physiol ; 164(4): 1636-48, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24715539

ABSTRACT

Drought is considered to cause oxidative stress, but the roles of oxidant-induced modifications in plant responses to water deficit remain obscure. Key unknowns are the roles of reactive oxygen species (ROS) produced at specific intracellular or apoplastic sites and the interactions between the complex, networking antioxidative systems in restricting ROS accumulation or in redox signal transmission. This Update discusses the physiological aspects of ROS production during drought, and analyzes the relationship between oxidative stress and drought from different but complementary perspectives. We ask to what extent redox changes are involved in plant drought responses and discuss the roles that different ROS-generating processes may play. Our discussion emphasizes the complexity and the specificity of antioxidant systems, and the likely importance of thiol systems in drought-induced redox signaling. We identify candidate drought-responsive redox-associated genes and analyze the potential importance of different metabolic pathways in drought-associated oxidative stress signaling.


Subject(s)
Droughts , Reactive Oxygen Species/metabolism , Antioxidants/metabolism , Oxidation-Reduction , Photosynthesis , Plant Growth Regulators/metabolism
18.
New Phytol ; 202(1): 145-160, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24299221

ABSTRACT

Oxidative stress responses are influenced by growth day length, but little is known about how this occurs. A combined reverse genetics, metabolomics and proteomics approach was used to address this question in Arabidopsis thaliana. A catalase-deficient mutant (cat2), in which intracellular oxidative stress drives pathogenesis-related responses in a day length-dependent manner, was crossed with a knockdown mutant for a specific type 2A protein phosphatase subunit (pp2a-b'γ). In long days (LD), the pp2a-b'γ mutation reinforced cat2-triggered pathogenesis responses. In short days (SD), conditions in which pathogenesis-related responses were not activated in cat2, the additional presence of the pp2a-b'γ mutation allowed lesion formation, PATHOGENESIS-RELATED GENE1 (PR1) induction, salicylic acid (SA) and phytoalexin accumulation and the establishment of metabolite profiles that were otherwise observed in cat2 only in LD. Lesion formation in cat2 pp2a-b'γ in SD was genetically dependent on SA synthesis, and was associated with decreased PHYTOCHROME A transcripts. Phosphoproteomic analyses revealed that several potential protein targets accumulated in the double mutant, including recognized players in pathogenesis and key enzymes of primary metabolism. We conclude that the cat2 and pp2a-b'γ mutations interact synergistically, and that PP2A-B'γ is an important player in controlling day length-dependent responses to intracellular oxidative stress, possibly through phytochrome-linked pathways.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/enzymology , Arabidopsis/immunology , Intracellular Space/metabolism , Oxidative Stress , Photoperiod , Protein Subunits/metabolism , Amino Acid Sequence , Antioxidants/metabolism , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis Proteins/genetics , Electrophoresis, Gel, Two-Dimensional , Flowers/genetics , Gas Chromatography-Mass Spectrometry , Gene Expression Regulation, Plant , Genotype , Indoles/metabolism , Molecular Sequence Data , Mutation/genetics , Phenotype , Phosphopeptides/chemistry , Phosphopeptides/metabolism , Phosphoproteins/chemistry , Phosphoproteins/metabolism , Plant Leaves/metabolism , Protein Phosphatase 2/genetics , Protein Phosphatase 2/metabolism , Proteome/metabolism , Proteomics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Salicylic Acid/metabolism , Thiazoles/metabolism
19.
Plant Cell Environ ; 37(5): 1114-29, 2014 May.
Article in English | MEDLINE | ID: mdl-24131360

ABSTRACT

Secondary metabolism plant glycosyltransferases (UGTs) ensure conjugation of sugar moieties to secondary metabolites (SMs) and glycosylation contributes to the great diversity, reactivity and regulation of SMs. UGT73B3 and UGT73B5, two UGTs of Arabidopsis thaliana (Arabidopsis), are involved in the hypersensitive response (HR) to the avirulent bacteria Pseudomonas syringae pv. tomato (Pst-AvrRpm1), but their function in planta is unknown. Here, we report that ugt73b3, ugt73b5 and ugt73b3 ugt73b5 T-DNA insertion mutants exhibited an accumulation of reactive oxygen species (ROS), an enhanced cell death during the HR to Pst-AvrRpm1, whereas glutathione levels increased in the single mutants. In silico analyses indicate that UGT73B3 and UGT73B5 belong to the early salicylic acid (SA)-induced genes whose pathogen-induced expression is co-regulated with genes related to cellular redox homeostasis and general detoxification. Analyses of metabolic alterations in ugt mutants reveal modification of SA and scopoletin contents which correlate with redox perturbation, and indicate quantitative modifications in the pattern of tryptophan-derived SM accumulation after Pst-AvrRpm1 inoculation. Our data suggest that UGT73B3 and UGT73B5 participate in regulation of redox status and general detoxification of ROS-reactive SMs during the HR to Pst-AvrRpm1, and that decreased resistance to Pst-AvrRpm1 in ugt mutants is tightly linked to redox perturbation.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/enzymology , Arabidopsis/microbiology , Disease Resistance/immunology , Glucosyltransferases/metabolism , Pseudomonas syringae/physiology , Secondary Metabolism , Arabidopsis/cytology , Arabidopsis/immunology , Arabidopsis Proteins/genetics , Ascorbic Acid/metabolism , Base Sequence , Cell Death , Computer Simulation , Disease Resistance/drug effects , Electrolytes/metabolism , Gene Expression Regulation, Plant/drug effects , Genes, Plant , Glucosyltransferases/genetics , Glutathione/metabolism , Glycosyltransferases/genetics , Glycosyltransferases/metabolism , Indoles/metabolism , Molecular Sequence Data , Mutation/genetics , Nucleotide Motifs/genetics , Oxidation-Reduction/drug effects , Plant Diseases/immunology , Plant Diseases/microbiology , Promoter Regions, Genetic/genetics , Pseudomonas syringae/drug effects , Pseudomonas syringae/growth & development , Reactive Oxygen Species/metabolism , Salicylic Acid/pharmacology , Scopoletin/metabolism , Secondary Metabolism/drug effects , Secondary Metabolism/genetics , Thiazoles/metabolism
20.
Plant J ; 69(4): 613-27, 2012 Feb.
Article in English | MEDLINE | ID: mdl-21985584

ABSTRACT

This work investigated the contribution of AtRbohD and AtRbohF to regulating defence-associated metabolism during three types of interaction: (i) incompatible and (ii) compatible interaction with Pseudomonas syringae; and (iii) intracellular oxidative stress in the catalase-deficient cat2 background. In all three cases, loss of function of either gene modulated the response of defence compounds. AtRbohF gene function was necessary for rapid and full induction of salicylic acid (SA) during compatible and incompatible interactions, and for resistance to virulent bacteria. Both artrboh mutations modulated the effects of intracellular ROS in the cat2 background, although the predominant effect was mediated by atrbohF. Loss of this gene function increased lesion formation in cat2 but uncoupled this effect from cat2-triggered induction of SA and camalexin, accumulation of glutathione and disease resistance, all of which were much lower in cat2 artbohF than in cat2. A detailed comparison of GC-TOF-MS profiles produced by the three interactions revealed considerable overlap between cat2 effects and those produced by bacterial infection in the wild-type background. Analysis of the impact of the two atrboh mutations on these profiles provided further evidence that AtRbohF interacts closely with intracellular oxidative stress to tune dynamic metabolic responses during infection. Thus, AtRbohF appears to be a key player not only in HR-related cell death but also in regulating metabolomic responses and resistance. Based on the results obtained during the three types of interaction, a model is proposed of how NADPH oxidases and intracellular ROS interact to determine the outcome of pathogen defence responses.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/physiology , Disease Resistance/physiology , Plant Diseases/immunology , Reactive Oxygen Species/metabolism , Stress, Physiological/physiology , Arabidopsis/genetics , Arabidopsis/immunology , Arabidopsis/microbiology , Arabidopsis Proteins/genetics , Catalase/metabolism , Cell Death , Gene Expression Regulation, Plant/physiology , Glutathione/metabolism , Indoles/metabolism , Metabolomics , Mutation , NADPH Oxidases/genetics , NADPH Oxidases/metabolism , Oxidation-Reduction , Oxidative Stress , Plant Diseases/microbiology , Plant Leaves/genetics , Plant Leaves/immunology , Plant Leaves/microbiology , Plant Leaves/physiology , Pseudomonas syringae/pathogenicity , Salicylic Acid/metabolism , Scopoletin/metabolism , Signal Transduction/physiology , Thiazoles/metabolism
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