Abstract
A highly anionic peroxidase was strongly suggested to be involved in the deposition of the aromatic domain of suberin. cDNA containing the coding region of the suberization-associated anionic peroxidase from potato has been cloned and sequenced. The deduced amino acid sequence of the peroxidase shows that it is an anionic protein with considerable homology to other peroxidases. The amino acid sequence of two tryptic peptides obtained from the anionic peroxidase purified from suberizing potato tuber slices matched exactly with two segments of the amino acid sequence deduced from the nucleotide sequence of the cloned cDNA. The identity of the cloned cDNA is further supported by hybrid-selected translation and immunological recovery of the product with antibodies prepared against the purified anionic peroxidase. This anionic peroxidase was barely detectable at 2 days after wounding, and reached a maximal level at 8 days after wounding. Using the cDNA for the anionic peroxidase as a probe, we showed that the mRNA for the enzyme was induced in suberizing potato. The mRNA levels increased from an undetectable level in control tuber tissue to a maximal level in suberizing tuber tissue aged for four days. In suberizing tomato fruit the peroxidase mRNA showed induction and the level reached a maximum in three days. Ine data suggest that the induction of the peroxidase by wounding is preceded by transcriptional activation of the peroxidase gene or by increased stabilization of the mRNA. The time course of increase in mRNA for the anionic peroxidase was consistent with its postulated role in suberization.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Agrawal VP, Kolattukudy PE: Biochemistry of suberization: ω-hydroxyacid oxidation in enzyme preparations from suberizing potato tuber disks. Plant Physiol 59: 667–672 (1977).
Benedum U, Baeuerle PA, Konecki DS, Frank R, Powell J, Mallet J, Huttner WB: The primary structure of bovine chromogranin A: a representative of a class of acidic secretory proteins common to a variety of peptidergic cells. EMBO J 5: 1495–1502 (1986).
Borchert R: Time course and spatial distribution of phenylalanine ammonia-lyase and peroxidase activity in wounded potato tuber tissue. Plant Physiol 62: 789–793 (1978).
Bradford MM: 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 (1976).
Chen EJ, Seeburg PH: Supercoil sequencing: a fast simple method for sequencing plasmid DNA. DNA 4: 165–170 (1985).
Chen J, Varner J: Isolation and characterization of cDNA clones for carrot extensin and a proline rich 33 kDa protein. Proc Natl Acad Sci USA 82: 4399–4403 (1985).
Chrestfield AM, Moore S, Stein WH: The preparation and enzymatic hydrolysis of reduced and S-carboxymethylated proteins. J Biol Chem 238: 622–627 (1963).
Cottle W, Kolattukudy PE: Abscisic acid stimulation of suberization: induction of enzymes and deposition of polymeric components and associated waxes in tissue cultures of potato tuber. Plant Physiol 70: 775–780 (1982).
Dean BB, Kolattukudy PE: Synthesis of suberin during wound-healing in jade leaves, tomato fruit, and bean pods. Plant Physiol 58: 411–416 (1976).
Espelie KE, Kolattukudy PE: Purification and characterization of an abscisic acid-inducible anionic peroxidase associated with suberization in potato (Solanum tuberosum). Arch Biochem Biophys 240: 539–545 (1985).
Espelie KE, Davis RW, Kolattukudy PE: Composition, ultrastructure and function of the cutin- and suberin-containing layers in the leaf, fruit peel, juice-sac and inner seed coat of grapefruit (Citrus paradisi Macfed.). Planta 149: 498–511 (1980).
Espelie KE, Franceschi VR, Kolattukudy PE: Immunocytochemical localization and time course of appearance of an anionic peroxidase associated with suberization in wound-healing potato tuber tissue. Plant Physiol. 81: 487–492 (1986).
Flurkey WH, Kolattukudy PE: In vitro translation of cutinase mRNA: evidence for a precursor form of an extracellular fungal enzyme. Arch Biochem Biophys 212: 154–161 (1981).
Heidecher G, Messing J: Structural analysis of plant genes. Ann Rev Plant Physiol 39: 439–466 (1986).
Henikoff S: Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28: 351–359 (1984).
Huynh TV, Young RA, Davis RW: Constructing and screening cDNA libraries in λgt10 and λgt11. In: Glover DM (ed) DNA Cloning, Vol. 1. IRL Press, Washington DC, pp. 19–27 (1984).
Kolattukudy PE: Biopolyester membranes of plants: cutin and suberin. Science 208: 990–1000 (1980).
Kolattukudy PE: Structure, biosynthesis and biodegradation of cutin and suberin. Ann Rev Plant Physiol 32: 539–567 (1981).
Lagrimini LM, Rothstein S: Tissue specificity of tobacco peroxidase isozymes and their induction by wounding and tobacco mosaic virus infection. Plant Physiol 84: 438–442 (1987).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1982.
Mazza G, Welinder KG: Covalent structure of turnip peroxidase 7. Cyanogen bromide fragments, complete structure and comparison to horesradish peroxidase c. Eur J Biochem 103: 481–489 (1980).
Mirels L, Bedi GS, Dickinson DP, Gross KW, Tabak LA: Molecular characterization of glutamic acid/glutamine-rich secretory proteins from rat submandibular glands. J Biol Chem 262: 7289–7297 (1987).
Okita TW, Eseene FC: Wheat storage proteins: isolation and characterization of the gliadin messenger RNAs. Plant Physiol 69: 834–839 (1982).
Pozuelo JM, Espelie KE, Kolattukudy PE: Magnesium deficiency results in increased suberization in endodermis and hypodermis of corn roots. Plant Physiol 74: 256–260 (1984).
Sanchez-Serrano J, Schmidt R, Schell J, Willmitzer C: Nucleotide sequence of proteinase inhibitor II encoding cDNA of potato (Solanum tuberosum) and its mode of expression. Mole Gen Genet 203: 15–20 (1986).
Sijmons PC, Kolattukudy PE, Bienfait HF: Iron deficiency decreases suberization in bean roots through a decrease in suberin-specific peroxidase activity. Plant Physiol 78: 115–120 (1985).
Thornburg RW, An G, Cleveland TG, Johnson R, Ryan CA: Wound-inducible expression of a potato inhibitor II-chloramphenicol acetyl transferase gene fusion in tobacco transgenic plants. Proc Natl Acad Sci USA 84: 744–748 (1987).
Welinder KG: Amino acid sequence studies of horseradish peroxidase. Amino and carboxyl termini, cyanogen bromide and tryptic fragments, a complete sequence, and some structural characteristics of horseradish peroxidase c. Eur J Biochem 96: 483–502 (1979).
Welinder KG: Plant peroxidases. Their primary, secondary and tertiary structures, and relation to cytochrome c peroxidase. Eur J Biochem 151: 497–504 (1985).
Wood D: Oligonucleotide screening of cDNA libraries. Focus 6 (3): 1–3 (1984).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Roberts, E., Kutchan, T. & Kolattukudy, P.E. Cloning and sequencing of cDNA for a highly anionic peroxidase from potato and the induction of its mRNA in suberizing potato tubers and tomato fruits. Plant Mol Biol 11, 15–26 (1988). https://doi.org/10.1007/BF00016010
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00016010