Reactive oxygen species (ROS) are produced as a normal product of plant cellular metabolism. Various environmental stresses lead to excessive production of ROS causing progressive oxidative damage and ultimately cell death. Despite their destructive activity, they are well-described second messengers in a variety of cellular processes, including conferment of tolerance to various environmental stresses. Whether ROS would serve as signaling molecules or could cause oxidative damage to the tissues depends on the delicate equilibrium between ROS production, and their scavenging. Efficient scavenging of ROS produced during various environmental stresses requires the action of several nonenzymatic as well as enzymatic antioxidants present in the tissues. In this paper, we describe the generation, sites of production and role of ROS as messenger molecules as well as inducers of oxidative damage. Further, the antioxidative defense mechanisms operating in the cells for scavenging of ROS overproduced under various stressful conditions of the environment have been discussed in detail.

https://downloads.hindawi.com/archive/2012/217037.pdf

Reactive Oxygen Species, Oxidative Damage, and Antioxidative Defense Mechanism in Plants under Stressful Conditions

Water deficits cause a reduction in the rate of photosynthesis. Exposure to mild water deficits, when relative water content (RWC) remains above 70%, primarily causes limitation to carbon dioxide uptake because of stomatal closure. With greater water deficits, direct inhibition of photosynthesis occurs. In both cases limitation of carbon dioxide fixation results in exposure of chloroplasts to excess excitation energy. Much of this can be dissipated by various photoprotective mechanisms. These include dissipation as heat via carotenoids, photorespiration, CAM idling and, in some species, leaf movements and other morphological features which minimize light absorption. The active oxygen species superoxide and singlet oxygen are produced in chloroplasts by photoreduction of Oxygen and energy transfer from triplet excited chlorophyll to oxygen, respectively. Hydrogen peroxide and hydroxyl radicals can form as a result of the reactions of superoxide. All these species are reactive and potentially damaging, causing lipid peroxidation and inactivation of enzymes. They are normally scavenged by a range of antioxidants and enzymes which are present in the chloroplast and other subcellular compartments. When carbon dioxide fixation is limited by water deficit, the rate of active oxygen formation increases in chloroplasts as excess excitation energy, not dissipated fay the photoprotective mechanisms, is used to form superoxide and singlet oxygen. However, photorespiratory hydrogen peroxide production in peroxisomes decreases. Increased superoxide can be detected by EPR (electron paramagnetic resonance) in chloroplasts from droughted plants. Stiperoxide formation leads to changes suggestive of oxidative damage including lipid peroxidation and a decrease in ascorbate. These changes are not, however, apparent until severe water deficits develop, and they could also be interpreted as secondary effects of water deficit-induced senescence or wounding. Non-lethal water deficits often result in increased activity of superoxide dismutase, glutathione reductase and monodehydroascorbate reductase. Increased capacity of these protective enzymes may be part of a general antioxidative response in plants involving regulation of protein synthesis or gene expression. Since the capacity of these enzymes is also increased by other treatments which cause oxidative damage, and which alter the balance between excitation energy input and carbon dioxide fixation such as low temperature and high irradiance, it is suggested that water deficit has the same effect. Light levels that are not normally excessive do become excessive and photoprotective/antioxidative systems are activated. Some of the photoprotective mechanisms themselves could result in active oxygen formation. Photoinhibitory damage also includes a component of oxidative damage. During normally-encountered degrees of water deficit the capacity of the antioxidant systems and their ability to respond to increased active oxygen generation may be sufficient to prevent overt expression of damage. Desiccation-tolerant tissues such as bryophytes, lichens, spores, seeds, some algae and a few vascular plant leaves can survive desiccation to below 30-40% RWC, A component of desiccation damage in seeds and bacteria is oxygen-dependent. Desiccation causes oxidation of glutathione, a major antioxidant, and appearance of a free radical signal detected by EPR in a number of tissues suggesting that oxidative damage has occurred. In photosynthetic cells damage may arise from photooxidation. Disruption of membrane-bound electron tranport systems in partially hydrated tissue could lead to reduction of oxygen to superoxide. Oxidation of lipids and sulphydryl groups may also occur in dry tissue. Tolerant cells recover upon rehydration and arc able to reduce their glutathione pool. Non-tolerant species go on to show further oxidative damage including lipid peroxidation. It is difficult to attribute this subsequent damage to the cause or effect of death. Embryos in seeds lose desiccation tolerance soon after imbibition. This is associated with membrane damage that has been attributed to superoxide-mediated deesterification of phospholipids and loss of lipophilic antioxidants. These effects are discussed in relation to other mechanisms involved in desiccation tolerance. Contents Summary 27 I. Introduction 28 II. Generation of active oxygen and defence mechanisms in plant cells 29 III. The effect of water deficit on photosynthesis 31 IV. Mechanisms for active oxygen generation during water deficit 36 V. Evidence for oxidative damage during water deficit 39 VI. Desiccation 47 VII. Conclusions 52 Acknowledgements 53 References 53.

The role of active oxygen in the response of plants to water deficit and desiccation.

Adaptation to environmental changes is crucial for plant growth and survival. However, the molecular and biochemical mechanisms of adaptation are still poorly understood and the signaling pathways involved remain elusive. Active oxygen species (AOS) have been proposed as a central component of plant adaptation to both biotic and abiotic stresses. Under such conditions, AOS may play two very different roles: exacerbating damage or signaling the activation of defense responses. Such a dual function was first described in pathogenesis but has also recently been demonstrated during several abiotic stress responses. To allow for these different roles, cellular levels of AOS must be tightly controlled. The numerous AOS sources and a complex system of oxidant scavengers provide the flexibility necessary for these functions. This review discusses the dual action of AOS during plant stress responses.

Dual action of the active oxygen species during plant stress responses.

The light-dependent generation of active oxygen species is termed photooxidative stress. This can occur in two ways: (1) the donation of energy or electrons directly to oxygen as a result of photosynthetic activity; (2) exposure of tissues to ultraviolet irradiation. The light-dependent destruction of catalase compounds the problem. Although generally detrimental to metabolism, superoxide and hydrogen peroxide may serve useful functions if rigorously controlled and compartmentalised. During photosynthesis the formation of active oxygen species is minimised by a number of complex and refined regulatory mechanisms. When produced, active oxygen species are eliminated rapidly by efficient antioxidative systems. The chloroplast is able to use the production and destruction of hydrogen peroxide to regulate the thermal dissipation of excess excitation energy. This is an intrinsic feature of the regulation of photosynthetic electron transport. Photoinhibition and photooxidation only usually occur when plants are exposed to stress. Active oxygen species are part of the alarm-signalling processes in plants. These serve to modify metabolism and gene expression so that the plant can respond to adverse environmental conditions, invading organisms and ultraviolet irradiation. The capacity of the antioxidative defense system is often increased at such times but if the response is not sufficient, radical production will exceed scavenging and ultimately lead to the disruption of metabolism. Oxidative damage arises in high light principally when the latter is in synergy with additional stress factors such as chilling temperatures or pollution. Environmental stress can modify the photooxidative processes in various ways ranging from direct involvement in light-induced free radical formation to the inhibition of metabolism that renders previously optimal light levels excessive. It is in just such situations that the capacity for the production of active oxygen species can exceed that for scavenging by the antioxidative defense systems. The advent of plant transformation, however, may have placed within our grasp the possibility of engineering greater stress tolerance in plants by enhancement of the antioxidative defence system.

Photooxidative stress in plants

: The unpolarized absorption and circular dichroism spectra of the fundamental vibrational transitions of the chiral molecule, 4-methyl-2-oxetanone, are calculated ab initio. Harmonic force fields are obtained using Density Functional Theory (DFT), MP2, and SCF methodologies and a 5S4P2D/3S2P (TZ2P) basis set. DFT calculations use the Local Spin Density Approximation (LSDA), BLYP, and Becke3LYP (B3LYP) density functionals. Mid-IR spectra predicted using LSDA, BLYP, and B3LYP force fields are of significantly different quality, the B3LYP force field yielding spectra in clearly superior, and overall excellent, agreement with experiment. The MP2 force field yields spectra in slightly worse agreement with experiment than the B3LYP force field. The SCF force field yields spectra in poor agreement with experiment.The basis set dependence of B3LYP force fields is also explored: the 6-31G* and TZ2P basis sets give very similar results while the 3-21G basis set yields spectra in substantially worse agreements with experiment. jg

Ab initio calculation of vibrational absorption and circular dichroism spectra using density functional force fields

http://www.diva-portal.org/smash/get/diva2:613422/FULLTEXT01

UV-B exposure, ROS, and stress: inseparable companions or loosely linked associates?

Influx of solar UV-B radiation (280-320 nm) will probably increase in the future due to depletion of stratospheric ozone. In plants, there are several targets for the deleterious UV-B radiation, especially the chloroplast. This review summarizes the early effects and responses of low doses of UV-B at the molecular level. The DNA molecules of the plant cells are damaged by UV due to the formation of different photoproducts, such as pyrimidine dimers, which in turn can be combatted by specialized photoreactivating enzyme systems. In the chloroplast, the integrity of the thylakoid membrane seems to be much more sensitive than the activities of the photosynthetic components bound within. However, the decrease of mRNA transcripts for the photosynthetic complexes and other chloroplast proteins are among very early events of UV-B damage, as well as protein synthesis. Other genes, encoding defence-related enzymes, e.g., of the flavonoid biosynthetic pathway, are rapidly up-regulated after commencement of UV-B exposure. Some of the cis-acting nucleotide elements and trans-acting protein factors needed to regulate the UV-induced expression of the parsley chalcone synthase gene are known.

UV-B damage and protection at the molecular level in plants.

Exposure of plants to UV-B radiation (280–320 nm) results in changes in expression of a large number of genes. Before UV-B radiation or light of other wavelengths can give rise to a cellular response, it has to be perceived by some kind of receptor, and the information transduced via a signalling pathway to the target molecules, be it proteins in the cytoplasm or the genetic material in the nucleus. The perception of low levels of UV-B probably occurs via a UV-B photoreceptor followed by several different signalling pathways. These pathways include second messengers such as calcium, kinases and the catalytic formation of reactive oxygen species. High levels of UV-B, on the other hand, probably cause cellular damage and oxidative stress, thus activating a general stress signal transduction pathway which leads to a response similar to that which occurs after pathogen attack and other stresses. Some of the genes identified so far as being regulated by UV-B encode proteins involved in the biosynthesis of protective pigments, DNA repair and antioxidative enzymes, photosynthetic genes, cell cycle genes, and stress genes induced by other types of stimuli (i.e. pathogenesis-related proteins and senescence-induced genes). In the light of the information obtained on components necessary for UV-B-induced changes in gene expression, we propose in this mini-review a working model for UV-B perception and signal transduction. This model also takes into account dosage differences for the observations, which imply a separation into UV-B-specific and more general stress signal transduction.

Molecular events following perception of ultraviolet‐B radiation by plants

L., cv. Greenfeast) were exposed to supplementary UV-B light (up to 8 days) starting on the 17th day after sowing. The effects of this exposure on photosynthesis and the content and activities of some chloroplast components of the mature leaves of these #ants were studied. (i) The total chorophyil content of pea leaves was approximately 4~o of that in the control leaves on the 8th day of UV-B exposure. Chlorophyll a levels decreased to a greater extent than the content of chlorophyll b. The decrease in carotenoids paralleled the decrease in chlorophyll b. (ii) On a chlorophyll basis, the contents of Photosystem I and cytochrome / were stable, whereas Photosystem II, ATP hydrolysis by the ATP synthase and the maximum ribulose-l,5-bisphosphate carboxylase (Rubisco) activity decreased by 55, 47 and 80%, respectively, when compared with the controls at the end of the 8-day illumination period. (iii) On a leaf-area basis, Photosystem I and cytochrome f content decreased by 58%, Photosystem II by 80%, ATP hydrolysis by 80%, and Rubisco activity by 90%, when compared with the controls. The in vivo activation of Rubisco was markedly increased in UV-B-treated pea leaves. The underlying mechanisms for these results are discussed. Introduction Solar radiation is essential for all plant fife. Light is not only the driving force of photosynthesis but it also triggers and regulates many morphogenic responses. Excessive photon flux density, however, is potentially harmful, if not lethal, for plants. Deleterious effects to photosynthesis may be caused by excess visible light (photoinhibition; Ref 1) and ultraviolet (UV) radiation [2,3]. Global ultraviolet irradiance on the earth's surface Abbreviations: ATPase, ATP hydrolytic activity of the ATP syn- thase/hydrolase; Bicine, N,N-bis(2-hydroxyethyl)glycine; Car, caro- tenoids; CF1, the catalytic component of the chloroplast ATP syn- thase/hydrolase; Chl, chlorophyll; C a, plants fixing carbon by the tricarboxylic acid pathway; D1, the herbicide-binding polypeptide of Photosystem II; F m, maximum chlorophyll fluorescence; Fo, initial chlorophyll fluorescence when Photosystem II reaction centres are open; F v, variable chlorophyll fluorescence (Fm-Fo); LHCII, the light-harvesting components associated with Photosystem II; PAR, photosynthetically active radiation; PS I, Photosystem I; PS II, Photo- system II; P700, the reaction centre chlorophyll a of Photosystem I; Rubisco, ribulose-l,5-bisphosphate carboxylase; RuP2, ribulose 1,5- bisphosphate; UV-B, ultraviolet-B radiation (280-320 nm); UV-C, ultaviolet-C radiation ( < 280 nm). Correspondence: W.S. Chow, CSIRO, Division of Plant Industry, GPO Box 1600, Canberra, ACT 2601, Australia. fluctuates with changes in solar altitude, cloud cover, atmospheric turbidity and stratospheric ozone con- centration. Little UV radiation below 290 nm is de- tected at the earth's surface due to the absorption of solar radiation in the 200-290 nm region by strato- spheric ozone. Whereas UV-C radiation (< 280 nm; e.g., the mercury lamp with mostly 254 nm emission) causes damage to photosynthesis, primarily to the water-splitting apparatus of Photosystem II (PS II), and by the destruction of plastoquinone, it is not relevant in the natural environment [4]. However, UV-B radiation (280-320 nm) is readily absorbed by nucleic acids, proteins, pigments and lipids to a lesser extent [2]. UV-B primarily inhibits photosynthesis, as well as damaging many other plant processes [2,3]. The effects of UV-B radiation have been studied on plant communities, individual plants, leaves, isolated chloroplasts or thylakoid membranes using a plethora of UV-B radiation procedures ranging from UV-B radi- ation only, to various light treatments supplemented with UV-B [2,3,5]. The present study was initiated to evaluate the effects of moderate levels of supplementary UV-B radiation on photosynthesis. The composition and function of chloroplasts from fully expanded ma- ture leaves of peas were compared following exposure of one-half of the pea plants to additional UV-B radia- tion from the 17th day after planting. Peas were shown 0005-2728/90/$03.50 © 1990 Elsevier Science Publishers B.V. (Biomedical Division)

Åke Strid

Papers

UV-B exposure, ROS, and stress: inseparable companions or loosely linked associates?

UV-B damage and protection at the molecular level in plants.

Molecular events following perception of ultraviolet‐B radiation by plants

Effects of supplementary ultraviolet-B radiation on photosynthesis

Effects of supplementary ultraviolet-B radiation on photosynthesis in Pisum sativum.