Abstract
The effects of ozone or sulfur dioxide on antioxidant enzymes were investigated in Arabidopsis thaliana. Plants were fumigated with 0.1–0.15 ppm ozone or sulfur dioxide up to about 1 week in an environment-controlled chamber. Both pollutants increased the activities of ascorbate peroxidase and guaiacol per-oxidase in leaves, but had little effect on the activities of superoxide dismutase, catalase, monodehydroascorbate reductase, dehydroascorbate reductase or glutathione reductase. Ozone was more effective than sulfur dioxide in increasing the activities of the peroxidases. Ascorbate peroxidase activity increased 1.8-fold without a lag period during fumigation with 0.1 ppm ozone, while guaiacol peroxidase activity increased 4.4-fold with a 1-day lag. Expression of the APX1 gene encoding cytosolic ascorbate peroxidase was further investigated. Its protein levels in leaves exposed to 0.1 ppm ozone for 4 or 8 days were 1.5-fold higher than in controls. Both ozone and sulfur dioxide elevated APX1 mRNA levels in leaves at 4 and 7 days, whereas at 1 day only ozone was effective. The induction of APX1 mRNA levels by ozone (3.4- to 4.1-fold) was more prominent than that by sulfur dioxide (1.6-to 2.6-fold). The APX1 mRNA level increased by day and decreased by night. Exposure of plants to 0.1 ppm ozone enhanced the APX1 mRNA level within 3 h, which showed a diurnal rhythm similar to that of the control. These results demonstrate that near-ambient concentrations of ozone as well as similar concentrations of sulfur dioxide can induce APX1 gene expression in A. thaliana.
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References
Alscher RG: Biosynthesis and antioxidant function of glutathione in plants. Physiol Plant 77: 457–464 (1989).
Asada K: Ascorbate peroxidase: a hydrogen peroxide-scavenging enzyme in plants. Physiol Plant 85: 235–241 (1992).
Asada K: Production and scavenging of active oxygen in chloroplasts. In: Scandalios JG (ed) Molecular Biology of Free Radical Scavenging Systems, pp. 173–192. Cold Spring Harbor Laboratory Press, Plainview (1992).
Asada K, Takahashi M, Nagate M: Assay and inhibitors of spinach superoxide dismutase. Agric Biol Chem 38: 471–473 (1974).
Bowler C, Van Montagu M, Inzé D: Superoxide dismutase and stress tolerance. Annu Rev Plant Physiol Plant Mol Biol 43: 83–116 (1992).
Castillo FJ, Greppin H: Extracellular ascorbic acid and enzyme activities related to ascorbic acid metabolism in Sedum album L. leaves after ozone exposure. Environ Exp Bot 28: 231–238 (1988).
Chance B, Maehly AC: Assay of catalases and peroxidases. In: Colowick SP, Kaplan NO (eds) Methods in Enzymology vol. 2: Preparation and Assay of Enzymes, pp. 764–775. Academic Press, San Diego (1955).
Craker LE: Ethylene production from ozone-injured plants. Environ Pollut 1: 299–304 (1971).
Darrall NM: The effect of air pollutants on physiological processes in plants. Plant Cell Environ 12: 1–30 (1989).
Eckey-Kaltenbach H, Ernst D, Heller W, Sandermann H Jr.: Biochemical plant responses to ozone. IV. Cross-induction of defensive pathways in parsley (Petroselinum crispum L.) plants. Plant Physiol 104: 67–74 (1994).
Ernst D, Schraudner M, Langebartels C, Sandermann H Jr.: Ozone-induced changes of mRNA levels of β-1,3-glucanase, chitinase and ‘pathogenesis-related’ protein 1b in tobacco plants. Plant Mol Biol 20: 673–682 (1992).
Foyer CH, Descourviéres P, Kunert KJ: Protection against oxygen radicals: an important defence mechanism studied in transgenic plants. Plant Cell Environ 17: 507–523 (1994).
Hérouart D, Bowler C, Willekens H, Van Camp W, Slooten L, Van Montagu M, Inzé D: Genetic engineering of oxidative stress resistance in higher plants. Phil Trans R Soc Lond B 342: 235–240 (1993).
Hossain MA, Nakano Y, Asada K: Monodehydroascorbate reductase in spinach chloroplasts and its participation in regeneration of ascorbate for scavenging hydrogen peroxide. Plant Cell Physiol 25: 385–395 (1984).
Kangasjärvi J, Talvinen J, Utriainen M, Karjalainen R: Plant defence systems induced by ozone. Plant Cell Environ 17: 783–794 (1994).
Kubo A, Saji H, Tanaka K,Kondo N: Genomic DNA structureof a gene encoding cytosolic ascorbate peroxidase from Arabidopsis thaliana. FEBS Lett 315: 313–317 (1993).
Kubo A, Saji H, Tanaka K, Tanaka K, Kondo N: Cloning and sequencing of a cDNA encoding ascorbate peroxidase from Arabidopsis thaliana. Plant Mol Biol 18: 691–701 (1992).
Lieberman M: Biosynthesis and action of ethylene. Annu Rev Plant Physiol 30: 533–591 (1979).
Lobarzewski J, Greppin H, Penel C, Gaspar Th (eds): Biochemical, Molecular, and Physiological Aspects of Plant Peroxidases. University of Geneva, Geneva (1991).
Maccarrone M, Veldink GA, Vliegenthart JFG: Thermal injury and ozone stress affect soybean lipoxygenases expression. FEBS Lett 309: 225–230 (1992).
Madamanchi NR, Alscher RG: Metabolic bases for differences in sensitivity of two pea cultivars to sulfur dioxide. Plant Physiol 97: 88–93 (1991).
Madamanchi NR, Donahue JL, Cramer CL, Alscher RG, Pedersen K: Differential response of Cu,Zn superoxide dismutases in two pea cultivars during a short-term exposure to sulfur dioxide. Plant Mol Biol 26: 95–103 (1994).
Malhotra SS, Khan AA: Biochemical and physiological impact of major pollutants. In: Treshow M (ed) Air Pollution and Plant Life, pp. 113–157. John Wiley, Chichester (1984).
Mehlhorn H: Ethylene-promoted ascorbate peroxidase activity protects plants against hydrogen peroxide, ozone and paraquat. Plant Cell Environ 13: 971–976 (1990).
Mehlhorn H, Cottam DA, Lucas PW, Wellburn AR: Induction of ascorbate peroxidase and glutathione reductase activities by interactions of mixtures of air pollutants. Free Radical Res Comm 3: 193–197 (1987).
Mittler R, Tel-Or E: Oxidative stress responses and shock protein in the unicellular cyanobacterium Synechococcus R2 (PCC-7942). Arch Microbiol 155: 125–130 (1991).
Mittler R, Zilinskas BA: Molecular cloning and characterization of a gene encoding pea cytosolic ascorbate peroxidase. J Biol Chem 267: 21802–21807 (1992).
Mittler R, Zilinskas BA: Regulation of pea cytosolic ascorbate peroxidase and other antioxidant enzymes during the progression of drought stress and following recovery from drought. Plant J 5: 397–405 (1994).
Miyake C, Asada K: Thylakoid-bound ascorbate peroxidase in spinach chloroplasts and photoreduction of its primary oxidation product monodehydroascorbate radicals in thylakoids. Plant Cell Physiol 33: 541–553 (1992).
Nakano Y, Asada H: Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22: 867–880 (1981).
Navari-Izzo F, Quartacci MF, Izzo R, Pinzino C: Degradation of membrane lipid components and antioxidant levels in Hordeum vulgare exposed to long-term fumigation with SO2. Physiol Plant 84: 73–79 (1992).
Pastori GM, Trippi VS: Antioxidative protection in a drought-resistant maize strain during leaf senescence. Physiol Plant 87: 227–231 (1993).
Pell EJ, Reddy GN: Oxidative stress and its role in air pollution toxicity. In: Pell EJ, Steffen KL (eds) Active Oxygen/Oxidative Stress and Plant Metabolism, pp. 67–75. American Society of Plant Physiologists, Rockville (1991).
Perl A, Perl-Treves R, Galili S, Aviv D, Shalgi E, Malkin S, Galun E: Enhanced oxidative-stress defense in transgenic potato expressing tomato Cu,Zn superoxide dismutases. Theor Appl Genet 85: 568–576 (1993).
Pitcher LH, Brennan E, Zilinskas BA: The antiozonant ethylenediurea does not act via superoxide dismutase induction in bean. Plant Physiol 99: 1388–1392 (1992).
Rao MV: Cellular detoxifying mechanisms determine the age dependent injury in tropical trees exposed to SO2. J Plant Physiol 140: 733–740 (1992).
Saji H, Tanaka K, Kondo N: Monoclonal antibodies to spinach ascorbate peroxidase and immunochemical detection of the enzyme in eight different plant species. Plant Sci 69: 1–9 (1990).
Sakaki T, Kondo N, Sugahara K: Breakdown of photosynthetic pigments and lipids in spinach leaves with ozone fumigation: role of active oxygens. Physiol Plant 59: 28–34 (1983).
Sharma YK, Davis KR: Ozone-induced expression of stress-related genes in Arabidopsis thaliana. Plant Physiol 105: 1089–1096 (1994).
Tanaka K: Tolerance to herbicides and air pollutants. In: Foyer CH, Mullineaux PM (eds) Causes of Photooxidative Stress and Amelioration of Defense Systems in Plants, pp. 365–378. CRC Press, Boca Raton, FL (1994).
Tanaka K, Kondo N, Sugahara K: Accumulation of hydrogen peroxide in chloroplasts of SO2-fumigated spinach leaves. Plant Cell Physiol 23: 999–1007 (1982).
Tanaka K, Masuda R, Sugimoto T, Omasa K, Sakaki T: Water deficiency-induced changes in the contents of defensive substances against active oxygen in spinach leaves. Agric Biol Chem 54: 2629–2634 (1990).
Tanaka K, Saji H, Kondo N: Immunological properties of spinach glutathione reductase and inductive biosynthesis of the enzyme with ozone. Plant Cell Physiol 29: 637–642 (1988).
Tanaka K, Sugahara K: Role of superoxide dismutase in defense against SO2 toxicity and an increase in superoxide dismutase activity with SO2 fumigation. Plant Cell Physiol 21: 601–611 (1980).
Tanaka K, Takeuchi E, Kubo A, Sakaki T, Haraguchi K, Kawamura Y: Two immunologically different isozymes of ascorbate peroxidase from spinach leaves. Arch Biochem Biophys 286: 371–375 (1991).
Thomsen B, Drumm-Herrel H, Mohr H: Control of the appearance of ascorbate peroxidase (EC 1.11.1.11) in mustard seedling cotyledons by photochrome and photo-oxidative treatments. Planta 186: 600–608 (1992).
Treshow M, Anderson FK: Plant Stress from Air Pollution. John Wiley, Chichester (1989).
Van Camp W, Willekens H, Bowler C, Van Montagu M, Inzé D, Reupold-Popp P, Sandermann H Jr, Langebartels C: Elevated levels of superoxide dismutase protect transgenic plants against ozone damage. Bio/technology 12: 165–168 (1994).
Willekens H, Langebartels C, Tiré C, Van Montagu M, Inzé D, Van Camp W: Differential expression of catalase genes in Nicotiana plumbaginifolia (L.). Proc Natl Acad Sci USA 91: 10450–10454 (1994).
Willekens H, Van Camp W, Van Montagu M, Inzé D, Langebartels C, Sandermann H Jr: Ozone, sulfur dioxide, and ultraviolet B have similar effects on mRNA accumulation of antioxidant genes in Nicotiana plumbaginifolia L. Plant Physiol 106: 1007–1014 (1994).
Yalpani N, Enyedi AJ, León J, Raskin I: Ultraviolet light and ozone stimulate accumulation of salicylic acid, pathogenesis-related proteins and virus resistance in tobacco. Planta 193: 372–376 (1994).
Zhong HH, Young JC, Pease EA, Hangarter RP, McClung CR: Interaction between light and the circadian clock in the regulation of CAT2 expression in Arabidopsis. Plant Physiol 104: 889–898 (1994).
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Kubo, A., Saji, H., Tanaka, K. et al. Expression of arabidopsis cytosolic ascorbate peroxidase gene in response to ozone or sulfur dioxide. Plant Mol Biol 29, 479–489 (1995). https://doi.org/10.1007/BF00020979
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DOI: https://doi.org/10.1007/BF00020979