Abstract
We report the isolation of a ripening-related apple cDNA which is complementary to a mRNA which may be involved in ethylene production. Poly(A)+ RNA was extracted from cortical tissue of ripe apple fruit (Malus domestica Borkh cv. Golden Delicious) and a cDNA library constructed in the plasmid vector pSPORT. The library was screened with pTOM13, a tomato cDNA clone thought to code for ACC oxidase in that fruit. An apple cDNA clone (pAP4) was isolated and sequenced. The 1182 bp cDNA insert includes an open reading frame of 942 bp, and shows strong homology with reported tomato and avocado sequences, both at the nucleic acid and amino acid levels. The polypeptide has a calculated molecular mass of 35.4 kDa and a calculated pI of 5.15. In apple cortical tissue, expression of pAP4-complementary RNA increased with ethylene production by the fruit during ripening. Expression was also enhanced in both ethylene-treated and wounded fruit.
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References
Adams DO, Yang SF: Ethylene biosynthesis: identification of 1-aminocyclopropane-1-carboxylic acid as an intermediate in the conversion of methionine to ethylene. Proc Natl Acad Sci USA 76: 170–174 (1979).
Apelbaum A, Burgoon AC, Anderson JD, Solomos T, Lieberman M: Some characteristics of the system converting 1-aminocyclopropane-1-carboxylic acid to ethylene. Plant Physiol 67: 80–84 (1981).
Bleeker AB, Kenyon WH, Somerville SC, Kende H: Use of monoclonal antibodies in the purification and characterization of 1-aminocyclopropane-1-carboxylate synthase, an enzyme in ethylene biosynthesis. Proc Natl Acad Sci USA 83: 7755–7759 (1986).
Cheverry JL, Pouliquen J, Le Guyader H, Marcellin P: Calcium regulation of exogenous and endogenous 1-aminocyclopropane-1-carboxylic acid bioconversion to ethylene. Physiol Plant 74: 53–57 (1988).
Devereux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395 (1984).
Dong JG, Yip WK, Yang SF: Monoclonal antibodies against apple 1-aminocyclopropane-1-carboxylate synthase. Plant Cell Physiol 32: 25–31 (1991).
Dong JG, Kim WT, Yip WK, Thompson GA, Li L, Bennett AB, Yang SF: Cloning of a cDNA encoding 1-aminocyclopropane-1-carboxylate synthase and expression of its mRNA in ripening apple fruit. Plant 185: 38–45 (1991).
Doyle JJ, Doyle JL: Isolation of plant DNA from fresh tissue. Focus 12: 13–15 (1990).
Engelman DM, Steitz TA, Goldman A: Identifying nonpolar transbilayer helices in amino acid sequences of membrane proteins. Annu Rev Biophys Chem 15: 321–353 (1986).
Fourney RM, Miyakoshi J, Day RSIII, Paterson MC: Northern blotting: efficient RNA staining and transfer. Focus 10: 5–7 (1988).
Hamilton AJ, Bouzayen M, Grierson D: Identification of a tomato gene for the ethylene-forming enzyme by expression in yeast. Proc Natl Acad Sci USA 88: 7434–7437 (1991).
Hamilton AJ, Lycett GW, Grierson D: Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature 346: 284–287 (1990).
Holdsworth MJ, Bird CR, Ray J, Schuch W, Grierson D: Structure and expression of an ethylene-related cDNA from tomato. Nucl Acids Res 15: 731–739 (1987).
Holdsworth MJ, Schuch W, Grierson D: Nucleotide sequence of an ethylene-related gene from tomato. Nucl Acids Res 15: 10600 (1987).
Holdsworth MJ, Schuch W, Grierson D: Organisation and expression of a wound/ripening-related small multigene family from tomato. Plant Mol Biol 11: 81–88 (1988).
Kende H: Enzymes of ethylene biosynthesis: Plant Physiol 91: 1–4 (1989).
Kyte J, Doolittle RF: A simple method for displaying the hydropathic character of a protein. J Mol Biol 157: 105–132 (1982).
Lay-Yee M, DellaPenna D, Ross GS: Changes in mRNA and protein during ripening in apple fruit (Malus domestica Borkh. cv. Golden Delicious). Plant Physiol 94: 850–853 (1990).
Lincoln JE, Cordes S, Read E, Fischer RL: Regulation of gene expression during Lycopersiocon esculentum (tomato) fruit development. Proc Natl Acad Sci USA 84: 2793–2797 (1987).
Lincoln JE, Fischer RL: Regulation of gene expression by ethylene. In: Verma DPS, Goldberg RB (eds) Plant Gene Research: Temporal and Spatial Regulation of Plant Genes. Springer-Verlag, New York (1988).
Mansour R, Latche A, Vaillant V, Pech J-C, Reid MS: Metabolism of 1-aminocyclopropane-1-carboxylic acid in ripening apple fruits. Physiol Plant 66: 495–502 (1986).
McGarvey DJ, Yu H, Christoffersen RE: Nucleotide sequence of a ripening-related cDNA from avocado fruit. Plant Mol Biol 15: 165–167 (1990).
Mol JNM, van der Krol AR, van Tunen AJ, van Blokland R, de Lange P, Stuitje AR: Regulation of plant gene expression by antisense RNA. FEBS Lett 268: 427–430 (1990).
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, pp. 1.94–1.95. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Sato T, Theologis A: Cloning the mRNA encoding 1-aminocyclopropane-1-carboxylate synthase, the key enzyme for ethylene biosynthesis in plants. Proc Natl Acad Sci USA 86: 6621–6625 (1989).
Southern EM: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517 (1975).
Spanu P, Reinhardt D, Boller T: Analysis and cloning of the ethylene-forming enzyme from tomato by functional expression of its mRNA in Xenopus laevis oocytes. EMBO J 10: 2007–2013 (1991).
Van der Straeten D, Van Wiemeersch L, Goodman HM, Van Montagu M: Cloning and sequence of two different cDNAs encoding 1-aminocyclopropane-1-carboxylate synthase in tomato. Proc Natl Acad Sci USA 87: 4859–4863 (1990).
Ververidis P, John P: Complete recovery in vitro of ethylene-forming enzyme activity. Phytochemistry 30: 725–727 (1991).
Yang SF: Biosynthesis of ethylene and its regulation. In: Friend J, Rhodes MJC (eds) Recent Advances in the Biochemistry of Fruits and Vegetables, pp. 89–106. Academic Press, London (1981).
Yang SF, Hoffman NE: Ethylene biosynthesis and its regulation in higher plants. Annu Rev Plant Physiol 35: 155–189 (1984).
Yip W-K, Dong J-G, Yang SF: Purification and characterisation of 1-aminocyclopropane-1-carboxylate synthase from apple fruits. Plant Physiol 95: 251–257 (1991).