Stress-triggered redox signalling: What’s in pROSpect?

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 H₂O₂ concentrations in the peroxisomes are provided, and based on the likely values in other locations such as chloroplasts, we conclude that much of the H₂O₂ 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

Thiol-based redox homeostasis and signaling

Plants imperatively have to cope with adverse conditions owing to their lack of mobility and to the high amounts of reactive oxygen species (ROS) generated from both respiration and photosynthetic metabolism. Although thiol redox homeostasis in plants is mainly preserved by the cellular glutathione pool, specific strategies have been adopted by the plant kingdom during evolution to manage these “extra” pro-oxidative conditions. Unlike human or yeast, plants generally possess a higher number of genes coding for antioxidant proteins, including protein families responsible of dithiol/disulfide exchange reactions. During the last decades, redox-dependent post-translational modifications of proteins proved to be pivotal to many cellular functions. In particular, this is critically important under some situations of environmental constraints taking into account the alterations and fine adjustment of the cellular redox status occurring during and after any biotic or abiotic stresses

The roles of reactive oxygen metabolism in drought: not so cut and dried

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

Towards a better understanding of antioxidant systems in plants under environmental constraints : experimental approaches and mechanistic modelling

Les voies métaboliques les plus importantes dans le contrôle du stress oxydant chez la plante restent à élucider. Celles liées au glutathion jouent un rôle important. Cependant, les réactions responsables de l'oxydation du glutathion (du GSH en GSSG) n'ont pas encore été clairement identifiées. L’analyse des données biochimiques, transcriptomiques et génétiques soulèvent des questions pour mieux comprendre comment la régulation redox liée au stress pourrait influer sur la signalisation hormonale chez les plantes. Par une approche de génétique inverse utilisant, notamment, le mutant photorespiratoire conditionnel cat2, nous avons étudié la réponse et l'importance fonctionnelle de trois voies potentielles, médiées par les glutathion S-transférases, les peroxirédoxines dépendant de la glutarédoxine et les déhydroascorbate réductases (DHARs) chez Arabidopsis. Ainsi, l'interaction entre les DHARs semble être nécessaire pour coupler les pools d'ascorbate et de glutathion lors d’un stress oxydant. En complément à l'approche expérimentale, une modélisation mécaniste a permis d'étudier la production de H2O2 et son métabolisme, en lien avec l'activité catalase et la voie ascorbate-glutathion. Le modèle révèle que la catalase et l'ascorbate peroxydase prennent en charge de concert le traitement de H2O2, y compris dans les conditions optimales de croissance. Nos simulations suggèrent que la disponibilité en NADPH peut déterminer l'oxydation du glutathion via la monodéshydroascorbate réductase. Nos résultats expérimentaux et le modèle cinétique valident que la sensibilité du statut du glutathion au stress oxydant constitue un senseur approprié des augmentations du H2O2.The most important metabolic pathways in the control of oxidative stress remain to be elucidated in plants. Those linked to glutathione play an important role. However, the reactions responsible for its oxidation have not been clearly identified. Here, analysis based on available biochemical, transcriptomic and genetic data emphasized likely important questions to be elucidated for a full understanding of how stress-related redox regulation might impinge on phytohormone-related signaling pathways. Using a reverse genetics approach and the photorespiratory conditional cat2 mutant, we studied the response and functional importance of three potential routes for glutathione oxidation pathways mediated by glutathione S-transferases, glutaredoxin dependent peroxiredoxins, and dehydroascorbate reductases (DHAR) in Arabidopsis during oxidative stress. Hence, interplay between different DHARs appears to be necessary to couple ascorbate and glutathione pools and to allow glutathione-related signaling during enhanced H2O2 metabolism. In addition to experimental work, modelling is another way to investigate H2O2 production and its metabolism related to catalase activity and ascorbate glutathione pathway. This approach led to major conclusions, that catalase and ascorbate peroxidase can share the load in H2O2 processing even in optimal growth conditions. Furthermore, simulations propose that NADPH availability may determine glutathione oxidation through its influence on monodehydroascorbate reduction. Taken together, experimental results and our kinetic model strengthen that the sensitivity of glutathione status to oxidative stress acts as a suitable sensor of increased H2O2

Vers une meilleure compréhension des systèmes antioxydants chez la plante face aux contraintes environnementales : approches expérimentales et modélisation mécaniste

The most important metabolic pathways in the control of oxidative stress remain to be elucidated in plants. Those linked to glutathione play an important role. However, the reactions responsible for its oxidation have not been clearly identified. Here, analysis based on available biochemical, transcriptomic and genetic data emphasized likely important questions to be elucidated for a full understanding of how stress-related redox regulation might impinge on phytohormone-related signaling pathways. Using a reverse genetics approach and the photorespiratory conditional cat2 mutant, we studied the response and functional importance of three potential routes for glutathione oxidation pathways mediated by glutathione S-transferases, glutaredoxin dependent peroxiredoxins, and dehydroascorbate reductases (DHAR) in Arabidopsis during oxidative stress. Hence, interplay between different DHARs appears to be necessary to couple ascorbate and glutathione pools and to allow glutathione-related signaling during enhanced H2O2 metabolism. In addition to experimental work, modelling is another way to investigate H2O2 production and its metabolism related to catalase activity and ascorbate glutathione pathway. This approach led to major conclusions, that catalase and ascorbate peroxidase can share the load in H2O2 processing even in optimal growth conditions. Furthermore, simulations propose that NADPH availability may determine glutathione oxidation through its influence on monodehydroascorbate reduction. Taken together, experimental results and our kinetic model strengthen that the sensitivity of glutathione status to oxidative stress acts as a suitable sensor of increased H2O2.Les voies métaboliques les plus importantes dans le contrôle du stress oxydant chez la plante restent à élucider. Celles liées au glutathion jouent un rôle important. Cependant, les réactions responsables de l'oxydation du glutathion (du GSH en GSSG) n'ont pas encore été clairement identifiées. L’analyse des données biochimiques, transcriptomiques et génétiques soulèvent des questions pour mieux comprendre comment la régulation redox liée au stress pourrait influer sur la signalisation hormonale chez les plantes. Par une approche de génétique inverse utilisant, notamment, le mutant photorespiratoire conditionnel cat2, nous avons étudié la réponse et l'importance fonctionnelle de trois voies potentielles, médiées par les glutathion S-transférases, les peroxirédoxines dépendant de la glutarédoxine et les déhydroascorbate réductases (DHARs) chez Arabidopsis. Ainsi, l'interaction entre les DHARs semble être nécessaire pour coupler les pools d'ascorbate et de glutathion lors d’un stress oxydant. En complément à l'approche expérimentale, une modélisation mécaniste a permis d'étudier la production de H2O2 et son métabolisme, en lien avec l'activité catalase et la voie ascorbate-glutathion. Le modèle révèle que la catalase et l'ascorbate peroxydase prennent en charge de concert le traitement de H2O2, y compris dans les conditions optimales de croissance. Nos simulations suggèrent que la disponibilité en NADPH peut déterminer l'oxydation du glutathion via la monodéshydroascorbate réductase. Nos résultats expérimentaux et le modèle cinétique valident que la sensibilité du statut du glutathion au stress oxydant constitue un senseur approprié des augmentations du H2O2

Analyzing the Function of Catalase and the Ascorbate–Glutathione Pathway in H₂O₂ Processing: Insights from an Experimentally Constrained Kinetic Model

International audienceSignificance: Plant stress involves redox signaling linked to reactive oxygen species such as hydrogen peroxide (H2O2), which can be generated at high rates in photosynthetic cells. The systems that process H2O2 include catalase (CAT) and the ascorbate–glutathione pathway, but interactions between them remain unclear. Modeling can aid interpretation and pinpoint areas for investigation. Recent Advances: Based on emerging data and concepts, we introduce a new experimentally constrained kinetic model to analyze interactions between H2O2, CAT, ascorbate, glutathione, and NADPH. The sensitivity points required for accurate simulation of experimental observations are analyzed, and the implications for H2O2-linked redox signaling are discussed. Critical Issues: We discuss several implications of the modeled results, in particular the following. (i) CAT and ascorbate peroxidase can share the load in H2O2 processing even in optimal conditions. (ii) Intracellular H2O2 concentrations more than the low μM range may rarely occur. (iii) Ascorbate redox turnover is largely independent of glutathione until ascorbate peroxidation exceeds a certain value. (iv) NADPH availability may determine glutathione redox status through its influence on monodehydroascorbate reduction. (v) The sensitivity of glutathione status to oxidative stress emphasizes its potential suitability as a sensor of increased H2O2. Future Directions: Important future questions include the roles of other antioxidative systems in interacting with CAT and the ascorbate–glutathione pathway as well as the nature and significance of processes that achieve redox exchange between different subcellular compartments. Progress in these areas is likely to be favored by integrating kinetic modeling analyses into experimentally based programs, allowing each approach to inform the other

Cytosolic and Chloroplastic DHARs Cooperate in Oxidative Stress-Driven Activation of the Salicylic Acid Pathway

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 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 H2O2 metabolism to glutathione oxidation; (3) DHAR-dependent glutathione oxidation influences redox-driven salicylic acid accumulation