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Enhanced oxidative-stress defense in transgenic potato expressing tomato Cu,Zn superoxide dismutases

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Summary

The two cDNAs coding for the cytosolic (cyt) and the chloroplast-located (chl) Cu,Zn superoxide dismutases (SODs) of tomato (Perl-Treves et al. 1988) were cloned into respective binary vectors and mobilized into Agrobacterium strains. Potato tuber discs were infected with either of the two agrobacterial strains and cultured on selective medium containing kanaymcin. The integration of either of the cyt or the chl SOD transgenes was verified by Southern-blot hybridization. The enzymatic activity of the additional tomato chl Cu,Zn SOD could be distinguished from endogenous SOD activity since the latter isozyme migrated faster on SOD-activity gels. Several transgenic potato lines harboring either the cyt or the chl SOD genes of tomato showed elevated tolerance to the superoxide-generating herbicide paraquat (methyl viologen). After exposure of shoots to paraquat, tolerance was recorded either by scoring symptoms visually or by measurements of photosynthesis using the photoacoustic method. Root cultures from transgenic lines that harbored the additional cyt Cu,Zn SOD gene of tomato were tolerant to methyl viologen up to 10-5 M; a lower tolerance was recorded in roots of transgenic lines that expressed the additional chl Cu,Zn SOD of tomato.

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References

  • An G (1986) Development of plant-promoter expression vectors and their use for analysis of differential activity of nopaline synthase promoter in transformed tobacco cells. Plant Physiol 81:86–91

    Google Scholar 

  • An G (1987) Binary vectors for plant transformation and promoter analysis. Methods Enzymol 153:292–305

    Article  CAS  Google Scholar 

  • Asada K, Takahashi M (1987) Production and scavenging of active oxygen in photosynthesis. In: Kyle DJ, Osmond CB, Arentzen CJ (eds) Photoinhibition. Elsevier Sciene Publishers. Amsterdam, pp 227–287

    Google Scholar 

  • Beauchamp CO, Fridovich I (1971) Superoxide dismutase. Improved assays and an assay applicable to acrylamide gel. Anal Biochem 44:276–287

    CAS  PubMed  Google Scholar 

  • Beyer W, Imaly J, Fridovich I (1991) Superoxide dismutases. Prog Nucleic Acid Res Molec Biol 40:221–253

    Google Scholar 

  • Bowler C, Alliote T, De Loose M, Van Montagu M, Inze D (1989) The induction of manganese superoxide dismutase in response to stress in Nicotiana plumbaginifolia. EMBO J 8:31–38

    CAS  PubMed  Google Scholar 

  • Bowler C, Slooten L, Vandenbranden S, De Rycke R, Botterman J, Sybesma C, Van Montagu M, Inze D (1991) Manganese superoxide dismutase can reduce cellular damage mediated by oxygen radicals in transgenic plants. EMBO J 10:1723–1732

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Bults G, Horwitz BA, Malkin S, Cahen D (1982) Photoacoustic measurements of photosynthetic activities in whole leaves — photochemistry and gas exchange. Biochim Biophys Acta 679:452–465

    Google Scholar 

  • Burke JJ, Gamble PE, Hatfield JL, Quisenberry JE (1985) Plant morphological and biochemical responses to field water deficit. I. Response of glutathione reductase activity and paraquat sensitivity. Plant Physiol 79:415–419

    Google Scholar 

  • Cannon RE, White JA, Scandalios JG (1987) Cloning of cDNA for maize superoxide dismutase 2 (SOD2). Proc Natl Acad Sci USA 84:179–183

    Google Scholar 

  • Decleire M, de Cat W, de Temmerman L, Baeten H (1984) Changes of peroxidase, catalase and superoxide dismutase activities in ozone-fumigated spinach leaves. J Plant Physiol 116:147–152

    Google Scholar 

  • Elroy-Stein O, Bernstein Y, Groner Y (1986) Overproduction of human Cu,Zn superoxide dismutase in transfected cells: extenuation of paraquat-mediated cytotoxicity and enhancement of lipid peroxidation. EMBO J 5:615–622

    Google Scholar 

  • Frank L (1985) Oxygen toxicity in eukaryotes. In: Oberley LW (ed) Superoxide dismutase, vol 3. CRC Press, Boca Raton, Florida, pp 1–43

    Google Scholar 

  • Furusawa I, Tanaka K, Thanutong P, Mizuguchi A, Yasaki M, Asada K (1984) Paraquat-resistant tobacco calluses with enhanced superoxide dismutase activity. Plant Cell Physiol 25:1247–1254

    Google Scholar 

  • Halliwell B (1984) Oxygen-derived species and herbicide action. Physiol Plant 15:21–24

    Google Scholar 

  • Halliwell B (1987) Oxidant and human disease: new concepts. FASEB J 1:358–364

    Google Scholar 

  • Havaux M, Canaani O, Malkin S (1986) Photosynthetic response of leaves to water stress, expressed by photoacoustic and related methods. I. Probing with the photoacoustic method as an indicator for water stress in vivo. Plant Physiol 82:827–833

    Google Scholar 

  • Havaux M, Canaani O, Malkin S (1987) Rapid screening for heat tolerance in Phaseolus species using the photoacoustic technique. Plant Sci 48:143–149

    Google Scholar 

  • Heiska E, Huttunen S (1987) Seasonal variation of superoxide dismutase activity in pine and spruce needles. Aquilo Ser Bot 25:39–47

    Google Scholar 

  • Imlay JA, Linn S (1988) DNA damage and oxygen radical toxicity Science 240:267–271

    Google Scholar 

  • Jansen MAK, Shaaltiel Y, Kazzes D, Canaani O, Malkin S, Gressel J (1989) Increased tolerance to photoinhibitory light in paraquat-resistant Conyza bonariensis measured by photoacoustic spectroscopy and 14CO2-fixation. Plant Physiol 91:1174–1178

    Google Scholar 

  • Jansen MAK, Malan C, Shaaltiel Y, Gressel J (1990) Mode of evolved photooxidant resistance to herbicides and xenobiotics. Z Naturforsch 45c:463–469

    Google Scholar 

  • Laemmli AK (1970) Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature 227:680–685

    PubMed  Google Scholar 

  • Matters GL, Scandalios JG (1986) Effect of the free radical-generating herbicide paraquat on the expression of the superoxide dismutase (SOD) genes of maize. Biochim Biophys Acta 882:29–38

    Google Scholar 

  • McCord JM, Fridovich I (1969) Superoxide dismutase, an enzymatic function for erythrocuprein (hemocuprein). J Biol Chem 60:6049–6055

    Google Scholar 

  • Monk LS, Davies HV (1989) Antioxidant status of the potato tuber and Ca2+ deficiency as a physiological stress. Physiol Plant 75:411–416

    Google Scholar 

  • Monk LS, Fagerstedt KV, Crawford RMM (1987) Superoxide dismutase as an anaerobic polypeptide — a key factor in recovery from oxygen deprivation in Iris pseudacous? Plant Physiol 85:1016–1020

    Google Scholar 

  • Monk LS, Fagerstedt KV, Crawford RMM (1989) Oxygen toxicity and superoxide dismutase as an antioxidant in physiological stress. Physiol Plant 76:456–459

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tissue cultures. Physiol Plant 15:473–497

    CAS  Google Scholar 

  • Nitsch JP (1969) Experimental androgenesis in Nicotiana. Phytomorphology 19:389–404

    Google Scholar 

  • Perl-Treves R (1990) Molecular cloning and expression of Cu,Zn superoxide dismutases in tomato. PhD thesis, The Weizmann Institute of Science, Rehovot, Israel

    Google Scholar 

  • Perl-Treves R, Galun E (1991) Developmental and light-induced regulation and stress-enhanced expression of two Cu,Zn superoxide dismutase genes in tomato. Plant Mol Biol 17:745–760

    Google Scholar 

  • Perl-Treves R, Nacmias B, Aviv D, Zeelon EP, Galun E (1988) Isolation of two cDNA clones from tomato containing two different superoxide dismutase sequences. Plant Mol Biol 11:609–623

    Google Scholar 

  • Perl-Treves R, Abu-Abied M, Magal N, Galun E, Zamir D (1990) Genetic mapping of tomato cDNA clones encoding the chloroplastic and the cytosolic isozymes of superoxide dismutase. Biochem Genet 28:543–552

    Google Scholar 

  • Phillips JP, Campbell SD, Michaud D, Charbonneau M, Hilliker A (1989) Null mutation of copper/zinc superoxide dismutase in Drosophila confers hypersensitivity to paraquat and reduced longevity. Proc Natl Acad Sci USA 86:2761–2765

    Google Scholar 

  • Pitcher LH, Brennan E, Hurley A, Dunsmuir P, Tepperman JM, Zilinskas BA (1991) Overproduction of petunia copper/zinc superoxide dismutase does not confer ozone tolerance in transgenic tobacco. Plant Physiol 97:452–455

    Google Scholar 

  • Polos E, Mikulas J, Szigeti Z, Matkovics B, Hai DQ, Parducz A, Lehoczki E (1988) Paraquat and atrazine co-resistance in Conyza canadensis L. Cronq. Pest Biochem Physiol 30:142–154

    Google Scholar 

  • Poulet P, Cahen D, Malkin S (1983) Photoacoustic detection of oxygen evolution from leaves — quantitative analysis by phase and amplitude measurements. Biochim Biophys Acta 724:433–446

    Google Scholar 

  • Reiss B, Sprengel R, Will H, Schaller H (1984) A new sensitive method for qualitative and quantitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30:211–218

    Google Scholar 

  • Rich PR, Bonner WD Jr (1978) The sites of superoxide anion generation in higher plant mitochondria. Arch Biochem Biophys 188:206–213

    Google Scholar 

  • Scioli JR, Zilinskas BA (1988) Cloning and characterization of a cDNA encoding the chloroplastic copper/zinc-superoxide dismutase from pea. Proc Natl Acad Sci USA 85:7661–7665

    Google Scholar 

  • Shaaltiel Y, Gressel J (1986) Multienzyme oxygen radical detoxifying system correlated with paraquat resistance in Conyza bonariensis. Pest Biochem Physiol 26:22–28

    Google Scholar 

  • Shaaltiel Y, Gressel J (1987) Kinetic analysis of resistance to paraquat in Conyza: evidence that paraquat transiently inhibits leaf chloroplast reaction in resistant plants. Plant Physiol 85:869–871

    Google Scholar 

  • Sheermann S, Bevan MW (1988) A rapid transformation method for Solanum tuberosum using Agrobacterium tumefaciens. Plant Cell Rep 7:13–16

    Google Scholar 

  • Szigeti Z, Polos E, Lehoczki E (1988) Fluorescence properties of paraquat-resistant Conyza leaves. In: Lichtenthaler HK (ed) Application of chlorophyll fluorescence. Kluwer Publishers, Dordrecht, pp 109–114

    Google Scholar 

  • Tepperman JM, Dunsmuir P (1990) Transformed plants with elevated levels of chloroplastic SOD are not more resistant to superoxide toxicity. Plant Mol Biol 14:501–511

    Google Scholar 

  • Tepperman JM, Katayama C, Dunsmuir P (1988) Cloning and nucleotide sequence of a petunia gene encoding a chloroplast-localized superoxide dismutase. Plant Mol Biol 11:871–882

    Google Scholar 

  • Tsang EWT, Bowler C, Herouart D, Van Camp W, Villarrod R, Genetello C, Van Montagu M, Inze D (1991) Differential regulation of superoxide dismutases in plants exposed to environmental stress. Plant Cell 3:783–792

    Article  CAS  PubMed  Google Scholar 

  • White DA, Zilinskas BA (1991) Nucleotide sequence of a complementary DNA encoding pea cytosolic copper/zinc superoxide dismutase. Plant physiol 96:1391–1392

    Google Scholar 

  • Yakir D, Rudich J, Bravdo BA, Malkin S (1986) Prolonged chilling under moderate light — effect on photosynthetic activities measured with the photoacoustic method. Plant Cell Environ 9:581–588

    Google Scholar 

  • Van Camp W, Bowler C, Villarrod R, Tsang EWT, Van Montagu M, Inze D (1990) Characterization of iron superoxide dismutase cDNAs from plants obtained by genetic complementation in Escherichia coli. Proc Natl Acad Sci USA 87:9903–9907

    Google Scholar 

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Authors and Affiliations

  1. Department of Plant Genetics, Weizmann Institute of Science, 76100, Rehovot, Israel

    A. Perl, R. Perl-Treves, S. Galili, D. Aviv & E. Galun

  2. Department of Biochemistry, The Weizmann Institute of Science, 76100, Rehovot, Israel

    E. Shalgi & S. Malkin

Authors
  1. A. Perl
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  2. R. Perl-Treves
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  3. S. Galili
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  4. D. Aviv
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  5. E. Shalgi
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  6. S. Malkin
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  7. E. Galun
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Communicated by R. Hagemann

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Perl, A., Perl-Treves, R., Galili, S. et al. Enhanced oxidative-stress defense in transgenic potato expressing tomato Cu,Zn superoxide dismutases. Theoret. Appl. Genetics 85, 568–576 (1993). https://doi.org/10.1007/BF00220915

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  • DOI: https://doi.org/10.1007/BF00220915

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