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Plant Science

Involvement of cytosolic ascorbate peroxidase and

Profile image of Gregorio Barba EspínGregorio Barba EspínProfile image of Jose A HernandezJose A Hernandez
https://doi.org/10.1093/JXB/ERQ432
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Abstract

In order to understand the role of cytosolic antioxidant enzymes in drought stress protection, transgenic tobacco (Nicotiana tabacum cv. Xanthi) plants overexpressing cytosolic Cu/Zn-superoxide dismutase (cytsod) (EC 1.15.1.1) or ascorbate peroxidase (cytapx) (EC 1.11.1.1) alone, or in combination, were produced and tested for tolerance against mild water stress. The results showed that the simultaneous overexpression of Cu/Znsod and apx or at least apx in the cytosol of transgenic tobacco plants alleviates, to some extent, the damage produced by water stress conditions. This was correlated with higher water use efficiency and better photosynthetic rates. In general, oxidative stress parameters, such as lipid peroxidation, electrolyte leakage, and H2O2 levels, were higher in non-transformed plants than in transgenic lines, suggesting that, at the least, overexpression of cytapx protects tobacco membranes from water stress. In these conditions, the activity of other antioxidant enzymes was induced in transgenic lines at the subcellular level. Moreover, an increase in the activity of some antioxidant enzymes was also observed in the chloroplast of transgenic plants overexpressing cytsod and/or cytapx. These results suggest the positive influence of cytosolic antioxidant metabolism on the chloroplast and underline the complexity of the regulation network of plant antioxidant defences during drought stress. Key words: Cytosolic ascorbate peroxidase, cytosolic superoxide dismutase, tobacco, water stress.

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Ghazi Badawi

Plant Science, 2004

A chimeric gene consisting of the coding sequence for cytosolic Cu/Zn-superoxide dismutases (SOD) from Oryza sativa fused to the chloroplast transit sequence from Arabidopsis thaliana glutathione reductase was used for generating transgenic tobacco plants. This construct was cloned into appropriate binary vector and mobilized into Agrobacterium tumefaciens C58C2. Tobacco leaf discs were infected with Agrobacterium and cultured on kanamycin selective medium. The integration and expression of Cu/Zn-SOD in tobacco genome was confirmed by Southern dot blot hybridization and SOD activity staining, respectively. Several transgenic lines were obtained and screened for SOD activity. The transgenic lines showed enhanced tolerance to the active oxygen-generating paraquat and sodium sulfite. Similarly, when net photosynthesis was measured, the first generation of the transgenic lines showed enhanced tolerance to salt, water, and PEG stresses, over the wild type. These results suggested that the overexpressed Cu/Zn-SOD enhances the chloroplast antioxidant system.

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Overexpression of an Arabidopsis Peroxisomal Ascorbate Peroxidase Gene in Tobacco Increases Protection Against Oxidative Stress
Randy Allen

Plant and Cell Physiology, 1999

The Arabidopsis gene APX3 that encodes a putative peroxisomal membrane-bound ascorbate peroxidase was expressed in transgenic tobacco plants. /l.PA'J-expressing lines had substantial levels of APX3 mRNA and protein. The H2O2 can be converted to more reactive toxic molecules, e.g. 'OH, if it is not quickly removed from plant cells. The expression of APX3 in tobacco could protect leaves from oxidative stress damage caused by aminotriazole which inhibits catalase activity that is found mainly in glyoxysomes and peroxisomes and leads to accumulation of H 2 O 2 in those organelles. However, these plants did not show increased protection from oxidative damage caused by paraquat which leads to the production of reactive oxygen species in chloroplasts. Therefore, protection provided by the expression of APX3 seems to be specific against oxidative stress originated from peroxisomes, not from chloroplasts, which is consistent with the hypothesis that APX3 is a peroxisomal membrane-bound antioxidant enzyme.

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Related topics

  • Chlorophyll Fluorescence
  • Drought and Water Stress
  • Antioxidative MEtabolism

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    The accumulation of endogenous proline induces changes in gene expression of several antioxidant enzymes in leaves of transgenic Swingle citrumelo
    Marília Kaphan Freitas Campos, Luiz Vieira

    Molecular Biology Reports, 2013

    Plant exposure to abiotic stresses leads to an accumulation of reactive oxygen species with the concomitant increase in antioxidant defense mechanisms. Previous studies showed that exogenous application of proline mitigate the deleterious effects caused by oxidative stress due to its ability to increase the activity of antioxidant enzymes. However, there are no reports of the effects of high endogenous accumulation of proline in the transcriptional pattern of antioxidant enzymes genes under normal conditions of water supply or in response to water deficit. Here, we show that isoforms of four antioxidant enzymes genes (Ascorbate peroxidase-APX, Catalase-CAT, Superoxide dismutase-SOD and Glutathione reductase-GR) were differentially regulated in leaves of Swingle citrumelo transgenic plants with high endogenous proline accumulation submitted to water deficits and also under normal water supply condition. Proline per se caused a twofold change in the transcription activity of APX1, APXcl, CAT2 and Cu/ZnSOD2, while during water deficit proline influenced mRNAs levels in APXs and Cu/ZnSODs isoforms, MnSODmit and GRcl. This study adds new information on the role of proline during drought conditions and, more important, without the potential confounding effects imposed by water deficiency. We showed that, in addition to its known effects on diverse plant physiological and biochemical processes, high endogenous proline can also acts as a regulatory/signalling molecule capable of altering the transcript levels of stress-related genes.

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    Genetic engineering of crops: a ray of hope for enhanced food security
    Sarvajeet Singh Gill

    Plant Signaling & Behavior, 2014

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    Ectopic expression of cytosolic superoxide dismutase and ascorbate peroxidase leads to salt stress tolerance in transgenic plums
    Pedro Díaz-vivancos

    Plant Biotechnology Journal, 2013

    These authors have contributed equally to this work.

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    Cadmium toxicity in Maize (Zea mays L.): consequences on antioxidative systems, reactive oxygen species and cadmium accumulation
    Shakeel Ahmad Anjum

    Environmental Science and Pollution Research, 2015

    Increased cadmium (Cd) accumulation in soils has led to tremendous environmental problems, with pronounced effects on agricultural productivity. Present study investigated the effects of Cd stress imposed at various concentrations (0, 75, 150, 225, 300, 375 μM) on antioxidant activities, reactive oxygen species (ROS), Cd accumulation, and productivity of two maize (Zea mays L.) cultivars viz., Run Nong 35 and Wan Dan 13. Considerable variations in Cd accumulation and in behavior of antioxidants and ROS were observed under Cd stress in both maize cultivars, and such variations governed by Cd were concentration dependent. Exposure of plant to Cd stress considerably increased Cd concentration in all plant parts particularly in roots. Wan Dan 13 accumulated relatively higher Cd in root, stem, and leaves than Run Nong 35; however, in seeds, Run Nong 35 recorded higher Cd accumulation. All the Cd toxicity levels starting from 75 μM enhanced H 2 O 2 and MDA concentrations and triggered electrolyte leakage in leaves of both cultivars, and such an increment was more in Run Nong 35. The ROS were scavenged by the enhanced activities of superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, and glutathione peroxidase in response to Cd stress, and these antioxidant activities were higher in Wan Dan 13 compared with Run Nong 35 at all Cd toxicity levels. The grain yield of maize was considerably reduced particularly for Run Nong 35 under different Cd toxicity levels as compared with control. The Wan Dan 13 was better able to alleviate Cd-induced oxidative damage which was attributed to more Cd accumulation in roots and higher antioxidant activities in this cultivar, suggesting that manipulation of these antioxidants and enhancing Cd accumulation in roots may lead to improvement in Cd stress tolerance.

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    Plant Signaling: Response to Reactive Oxygen Species
    Tanveer Bilal

    Plant signaling: Understanding the molecular crosstalk, 2013

    It is noteworthy to mention how the last 20 years have modified the concept of signalling in plants, especially the molecular crosstalk associated with it. Plants have the ability to show remarkable developmental plasticity to sustain in a continually changing environment. In response to various environmental stresses such as drought, salinity, metal toxicity, temperature and pathogens, plants defend themselves by developing some special defence mechanisms. Plants recognise these environmental signals with the help of some membrane protein sensors and then transduce these signals to the nucleus which ultimately stimulates various transcription factors and genes to form the product that ultimately leads to plant adaptation and assists the plant to sustain and surpass the adverse conditions. Amongst the environmental factors which are involved in signalling is the reactive oxygen species (ROS) generated during cell metabolism. ROS are spontaneously produced in the cell enzymatically through the action of various soluble membrane-bound enzymes and nonenzymatically by autoxidation reactions. Some of these ROS (e.g. superoxide dismutase, hydrogen peroxide and nitric oxide) are physiologically useful and in fact necessary for life but can also be harmful if present in excess or in inappropriate amounts. Current research in this regard focuses more on the development of transgenic plants with enhanced tolerance to ROS by using genetic approaches and analytical techniques. In particular nitric oxide (NO), a reactive radical, may be involved in the defence mediated by the ROS such as defence gene activation, hypersensitive response cell death and phytoalexin biosynthesis. By using biotechnological approaches NO together with ROS activates a stronger response and tolerance to various stresses in plants.

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