Christine H. Foyer

The presence of glutathione and glutathione reductase in chloroplasts: A proposed role in ascorbic acid metabolism.

Both glutathione and an NADPH-dependent glutathione reductase are present in spinach (Spinacia oleracea L.) chloroplasts. It is proposed that glutathione functions to stabilise enzymes of the Calvin cycle, and it may also act to keep ascorbic acid in chloroplasts in the reduced form.

Redox Homeostasis and Antioxidant Signaling: A Metabolic Interface between Stress Perception and Physiological Responses

Low molecular weight antioxidants, such as ascorbate, glutathione, and tocopherol, are information-rich redox buffers that interact with numerous cellular components. In addition to crucial roles in defense and as enzyme cofactors, cellular antioxidants influence plant growth and development by modulating processes from mitosis and cell elongation to senescence and death (De Pinto and De Gara, 2004; Potters et al., 2004; Tokunaga et al., 2005). Most importantly, antioxidants provide essential information on cellular redox state, and they influence gene expression associated with biotic and abiotic stress responses to maximize defense. Growing evidence suggests a model for redox homeostasis in which the reactive oxygen species (ROS)–antioxidant interaction acts as a metabolic interface for signals derived from metabolism and from the environment. This interface modulates the appropriate induction of acclimation processes or, alternatively, execution of cell death programs.

Ascorbate and Glutathione: The Heart of the Redox Hub

The discovery that there is a close relationship between ascorbate and glutathione dates from soon after the characterization of the chemical formulae of the two molecules ([Szent-Gyorgyi, 1931][1]; [Hopkins and Morgan, 1936][2]). Similarly, it has long been known that thylakoids can generate

Glutathione in plants: an integrated overview.

Plants cannot survive without glutathione (γ-glutamylcysteinylglycine) or γ-glutamylcysteine-containing homologues. The reasons why this small molecule is indispensable are not fully understood, but it can be inferred that glutathione has functions in plant development that cannot be performed by other thiols or antioxidants. The known functions of glutathione include roles in biosynthetic pathways, detoxification, antioxidant biochemistry and redox homeostasis. Glutathione can interact in multiple ways with proteins through thiol-disulphide exchange and related processes. Its strategic position between oxidants such as reactive oxygen species and cellular reductants makes the glutathione system perfectly configured for signalling functions. Recent years have witnessed considerable progress in understanding glutathione synthesis, degradation and transport, particularly in relation to cellular redox homeostasis and related signalling under optimal and stress conditions. Here we outline the key recent advances and discuss how alterations in glutathione status, such as those observed during stress, may participate in signal transduction cascades. The discussion highlights some of the issues surrounding the regulation of glutathione contents, the control of glutathione redox potential, and how the functions of glutathione and other thiols are integrated to fine-tune photorespiratory and respiratory metabolism and to modulate phytohormone signalling pathways through appropriate modification of sensitive protein cysteine residues.

COMMON COMPONENTS, NETWORKS, AND PATHWAYS OF CROSS-TOLERANCE TO STRESS. THE CENTRAL ROLE OF REDOX AND ABSCISIC ACID-MEDIATED CONTROLS

The vigor and responsiveness of plants to environmental stress result from the constant re-adjustment of physiology and metabolism throughout the life cycle within the framework of the genetic background. Plants have developed unique strategies for responding to ever-changing environmental conditions, exhaustively monitoring their surroundings and adjusting their metabolic systems to maintain homeostasis. The severity of stress, the genetic background of the plant, and its individual history determine everyday survival or death. These factors dictate the destiny of any individual. The genomeenvironment interaction is, therefore, an essential focus for the elucidation of the nature of the phenotypic variation leading to the successful response of plants to environmental cues. Plants acclimate to biotic and abiotic stresses by triggering a cascade or network of events that starts with stress perception and ends with the expression of a battery of target genes. The key components of the stress-response relationship are illustrated in Fig. 1. These are stress stimulus, signals, transducers, transcription regulators, target genes, and stress responses, including morphological, biochemical, and physiological changes. In evolutionary terms, components that are near to the end of the stressresponse cascade are not predicted to be the ones whose actions significantly affect the operation of other genes. However, factors that act at early stages are critical for other cell functions. Plants make use of common pathways and components in the stressresponse relationship. This phenomenon, which is known as cross-tolerance, allows plants to adapt/ acclimate to a range of different stresses after exposure to one specific stress. The major focus of this review, therefore, concerns the basic features of signaling that underpin cross-tolerance and result from the action of common elements, which are likely to occur early in the stress response cascade. First, using drought and chilling as examples, we explore the evidence for common signals and elements that confer cross-tolerance. Second, we highlight the importance of “redox signals” in such networks and discuss the evidence to date for the existence of such pathways in plants. The elucidation of common components has enormous potential and has, therefore, become a priority in research and breeding programs aimed at improving plant stress tolerance.

Understanding Oxidative Stress and Antioxidant Functions to Enhance Photosynthesis

Photosynthesis is a well-established source of reactive oxygen species (ROS) in plants. The photosynthetic electron transport chain (PET) operates in an aerobic environment; thus, regulatory systems are required to minimize ROS production. Moreover, an efficient antioxidant network is also essential

Leaf Mitochondria Modulate Whole Cell Redox Homeostasis, Set Antioxidant Capacity, and Determine Stress Resistance through Altered Signaling and Diurnal Regulation

To explore the role of plant mitochondria in the regulation of cellular redox homeostasis and stress resistance, we exploited a Nicotiana sylvestris mitochondrial mutant. The cytoplasmic male-sterile mutant (CMSII) is impaired in complex I function and displays enhanced nonphosphorylating rotenone-insensitive [NAD(P)H dehydrogenases] and cyanide-insensitive (alternative oxidase) respiration. Loss of complex I function is not associated with increased oxidative stress, as shown by decreased leaf H2O2 and the maintenance of glutathione and ascorbate content and redox state. However, the expression and activity of several antioxidant enzymes are modified in CMSII. In particular, diurnal patterns of alternative oxidase expression are lost, the relative importance of the different catalase isoforms is modified, and the transcripts, protein, and activity of cytosolic ascorbate peroxidase are enhanced markedly. Thus, loss of complex I function reveals effective antioxidant crosstalk and acclimation between the mitochondria and other organelles to maintain whole cell redox balance. This reorchestration of the cellular antioxidative system is associated with higher tolerance to ozone and Tobacco mosaic virus.

Leaf Vitamin C Contents Modulate Plant Defense Transcripts and Regulate Genes That Control Development through Hormone Signaling

Vitamin C deficiency in the Arabidopsis mutant vtc1 causes slow growth and late flowering. This is not attributable to changes in photosynthesis or increased oxidative stress. We have used the vtc1 mutant to provide a molecular signature for vitamin C deficiency in plants. Using statistical analysis, we show that 171 genes are expressed differentially in vtc1 compared with the wild type. Many defense genes are activated, particularly those that encode pathogenesis-related proteins. Furthermore, transcript changes indicate that growth and development are constrained in vtc1 by the modulation of abscisic acid signaling. Abscisic acid contents are significantly higher in vtc1 than in the wild type. Key features of the molecular signature of ascorbate deficiency can be reversed by incubating vtc1 leaf discs in ascorbate. This finding provides evidence that many of the observed effects on transcript abundance in vtc1 result from ascorbate deficiency. Hence, through modifying gene expression, vitamin C contents not only act to regulate defense and survival but also act via phytohormones to modulate plant growth under optimal conditions.

Overexpression of Glutathione Reductase but Not Glutathione Synthetase Leads to Increases in Antioxidant Capacity and Resistance to Photoinhibition in Poplar Trees

A poplar hybrid, Populus tremula x Populus alba, was transformed with the bacterial genes for either glutathione reductase (GR) (gor) or glutathione synthetase (GS) (gshll). When the gor gene was targeted to the chloroplasts, leaf GR activities were up to 1000 times greater than in all other lines. In contrast, targeting to the cytosol resulted in 2 to 10 times the GR activity. GR mRNA, protein, and activity levels suggest that bacterial GR is more stable in the chloroplast. When the gshll gene was expressed in the cytosol, GS activities were up to 100 times greater than in other lines. Overexpression of GR or GS in the cytosol had no effect on glutathione levels, but chloroplastic-GR expression caused a doubling of leaf glutathione and an increase in reduction state. The high-chloroplastic-GR expressors showed increased resistance to photoinhibition. The herbicide methyl viologen inhibited CO2 assimilation in all lines, but the increased leaf levels of glutathione and ascorbate in the high-chloroplastic-GR expressors persisted despite this treatment. These results suggest that overexpression of GR in the chloroplast increases the antioxidant capacity of the leaves and that this improves the capacity to withstand oxidative stress.

Photorespiratory Metabolism: Genes, Mutants, Energetics, and Redox Signaling

Photorespiration is a high-flux pathway that operates alongside carbon assimilation in C(3) plants. Because most higher plant species photosynthesize using only the C(3) pathway, photorespiration has a major impact on cellular metabolism, particularly under high light, high temperatures, and CO(2) or water deficits. Although the functions of photorespiration remain controversial, it is widely accepted that this pathway influences a wide range of processes from bioenergetics, photosystem II function, and carbon metabolism to nitrogen assimilation and respiration. Crucially, the photorespiratory pathway is a major source of H(2)O(2) in photosynthetic cells. Through H(2)O(2) production and pyridine nucleotide interactions, photorespiration makes a key contribution to cellular redox homeostasis. In so doing, it influences multiple signaling pathways, particularly those that govern plant hormonal responses controlling growth, environmental and defense responses, and programmed cell death. The potential influence of photorespiration on cell physiology and fate is thus complex and wide ranging. The genes, pathways, and signaling functions of photorespiration are considered here in the context of whole plant biology, with reference to future challenges and human interventions to diminish photorespiratory flux.