Abstract
A Scots pine (Pinus sylvestris L.) cDNA library was screened with two heterologous cDNA probes (P31 and T10) encoding cytosolic and chloroplastic superoxide dismutases (SOD) from tomato. Several positive clones for cytosolic and chloroplastic superoxide dismutases were isolated, subcloned, mapped and sequenced. One of the cDNA clones (PS3) had a full-length open reading frame of 465 bp corresponding to 154 amino acid residues and showed approximately 85% homology with the amino acid sequences of angiosperm cytosolic SOD counterparts. Another cDNA clone (PST13) was incomplete, but encoded a putative protein with 93% homology to pea and tomato chloroplastic superoxide dismutase. The derived amino acid sequence from both cDNA clones matched the corresponding N-terminal amino acid sequence of the purified mature SOD isozymes. Northern blot hybridizations showed that, cytosolic and chloroplastic CuZn-SOD are expressed at different levels in Scots pine organs. Sequence data and Southern blot hybridization confirm that CuZn-SODs in Scots pine belong to a multigene family. The results are discussed in relation to earlier observations of CuZn-SODs in plants.
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References
Asada K: Superoxide dismutase. In: Otsuka S, Yamanaka T (eds) Metalloproteins, pp. 331–341 Elsevier, Amsterdam (1988).
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K: Screening of recombinant DNA libraries. In: Current Protocols of Molecular Biology. Green Publishing Associates/Wiley-Interscience, New York (1989).
Baum JA, Scandalios JG: Multiple genes controlling superoxide dismutase expression in maize. J Hered 73: 95–100 (1982).
Cannon RE, White JA, Scandalios JG: Cloning of cDNA for maize superoxide dismutase 2 (SOD2). Proc Natl Acad Sci USA 84: 179–183 (1987).
Cannon RE, Scandalios JG: Two cDNAs encode two nearly identical Cu/Zn superoxide dismutase proteins in maize. Mol Gen Genet 219: 1–8 (1989).
Chen EY, Seeburg PH: Supercoil Sequencing: A fast and simple method for sequencing plasmid DNA. DNA 4: 165–170 (1985).
Devereux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395 (1984).
Fridrovich I: Superoxide dismutases. Adv Enzymol 58: 62–97 (1986).
Hassan HM, Scandalios JG: Superoxide dismutases in aerobic organisms. In: Alscher RG, Cumming JR (eds) Stress Responses in Plants: Adaptation and Acclimation Mechanisms, pp. 175–199. Wiley-Liss, New York (1990).
Ingestad T: Mineral nutrient requirements for Pinus sy.vestris and Picea abies seedlings. Physiol Plant 45: 373–380 (1979).
Joshi CP: Putative polyadenylation signals in nuclear genes of higher plants: A compilation and anlysis. Nucl Acids Res 15: 9627–9641 (1987).
Kanematso S, Asada K: Characteristic amino acid sequences of chloroplast and cytosol isozymes of CuZn-superoxide dismutase in spinach, rice and horsetail. Plant Cell Physiol 31: 99–112 (1990).
Kitagava Y, Tsunasawa S, Tanaka N, Katsube Y, Sakiyama F, Asada K: Amino acid sequence of copper, zinc-superoxide dismutase from spinach leaves. J Biochem 99: 1289–1298 (1986).
Kwiatowski JA, Kaniuga Z: Isolation and characterization of cytosolic and chloroplast isoenzymes of CuZn-superoxide dismutase from tomato leaves and their relationship to other CuZn-superoxide dismutase. Biochim Biophys Acta 874: 99–115 (1986).
Oseki H, Umesono K, Inokuchi H: The chloroplast genome of plants: a unique origin. Genome 31: 169–174. (1989).
Palmer JG: Evolution of chloroplast and mitochondrial DNA in plants and algae. In: MacIntyre RJ (ed), Molecular Evolutionary Genetics, pp. 131–240. Plenum Press, New York/London (1985).
Perl-Treves R, Nacimas B, Aviv D, Zeelon EP, Galun E: Isolation of two cDNA clones from tomato containing two different superoxide dismutase sequences. Plant Mol Biol 11: 609–624 (1988).
Rabinowich HD, Fridrovich I: Superoxide radicals, superoxide dismutase and oxygen toxicity in plants. Rev Photochem Photobiol 37: 679–690 (1983).
Sambrook J, Fritsch T, Maniatis F: Molecular Cloning: A laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Sanger F, Nicklen S and Coulson AR: DNA sequencing with chainterminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Scioli JR, Zilinskas BA: Cloning and characterization of cDNA encoding the chloroplastic copper/zinc-superoxide dismutase from pea. Proc Natl Acad Sci USA 85: 7661–7665 (1988).
Steffens GJ, Michelson AM, Ötting F, Puget K, Strassburger W, Flohé L: Primary structure of Cu-Zn superoxide dismutase of Brassica oleracea proves homology with corresponding enzymes of animals, fungi and procaryotes. Biol Chem Hoppe-Seyler 367: 1007–1016 (1986).
Szmidt AE, Lidholm J, Hällgren J-E: DNA extraction and preliminary characterisation of chloroplast DNA from Pinus sylvestris and Pinus contorta. In: Lindgren D (ed) Provenances and Forest Tree Breeding for High Latitudes, pp. 269–280. Proceedings of the Frans Kempe symposium, Umeå, 10–11 June (1986).
Tepperman J, Katayama C, Dunsmuir P: Cloning and nucleotide sequence of a petunia gene encoding a chloroplast-localized superoxide dismutase. Plant Mol Biol 11: 871–872 (1988).
Whitmore FW, Kriebel HB: Expression of a gene in Pinus strobus ovules associated with fertilization and early embryo development. Can J Forest Res 17: 408–412 (1987).
Wingsle G, Gardeström P, Hällgren J-E, Karpinski S: Isolation, purification and subcellular localization of isozymes of superoxide dismutase from Scots pine (Pinus sylvestris L.) needles. Plant Physiol 95: 21–28 (1991).
Young RG, Davis RW: Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci USA 80: 1194–1198 (1983).
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Karpinski, S., Wingsle, G., Olsson, O. et al. Characterization of cDNAs encoding CuZn-superoxide dismutases in Scots pine. Plant Mol Biol 18, 545–555 (1992). https://doi.org/10.1007/BF00040670
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DOI: https://doi.org/10.1007/BF00040670