Journal of Plant Research

, Volume 108, Issue 4, pp 463–468 | Cite as

Purification of cationic peroxidases bound ionically to the cell walls from the roots ofZinnia elegans

  • Yasushi Sato
  • Munetaka Sugiyama
  • Takashi Takagi
  • Hiroo Fukuda
Original Articles


Cell wall-bound and tracheary element-specific peroxidase isoenzymes, designated P5A and P5B, were shown previously to be associated with lignification during the differentiation into tracheary elements of single cells isolated from the mesophyll ofZinnia elegans (Satoet al. Planta 189: 584–589, 1993; Planta 196: 141–147, 1995). Isoenzymes corresponding to P5 (RP5A and RP5B) were present at a relatively high level in the roots ofZinnia elegans. These isoenzymes were purified from theZinnia roots by several column-chromatographic steps. Both RP5A and RP5B had molecular masses of 35 kDa. Purified RP5A and RP5B were cleaved by CNBr and the partial amino acid sequences of these isoenzymes were determined.

Key words

Amino acid sequence Lignification Peroxidase isoenzymes Tracheary element differentiation Zinnia elegans 



3-cyclohexylaminopropanesulfonic acid


Coomassie Brilliant Blue R-250






polyacrylamide gel electrophoresis


polyvinylidene difluoride


sodium dodecyl sulfate


tracheary element


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  1. Abeles, F.B. andBiles, C.L. 1991. Characterization of peroxidases in lignifying peach fruit endocarp. Plant Physiol.95: 269–273.Google Scholar
  2. Buffard, D., Breda, C., van Huystee, R.B., Asemota, O., Pierre, M., Ha, D.B.D. andEsnault, R. 1990. Molecular cloning of complementary DNAs encoding two cationic peroxidases from cultivated peanut cells. Proc. Natl. Acad. Sci. USA87: 8874–8878.PubMedGoogle Scholar
  3. Church, D.L. andGalston, A.W. 1988. 4-Coumarate: coenzyme A ligase and isoperoxidase expression inZinnia mesophyll cells induced to differentiate into tracheary elements. Plant Physiol.88: 679–684.PubMedGoogle Scholar
  4. Ferrer, M.A., Pedreño, M.A., Muñoz, R. andRos Barceló, A. 1990. Oxidation of coniferyl alcohol by cell wall peroxidases at the expense of indole-3-acetic acid and O2; a model for lignification of plant cell walls in the absence of H2O2. FEBS Lett.276: 127–130.CrossRefPubMedGoogle Scholar
  5. Fukuda, H. andKomamine, A. 1980. Establishment of an experimental system for the study of tracheary element differentiation from single cells isolated from the mesophyll ofZinnia elegans. Plant Physiol.65: 57–60.Google Scholar
  6. Fukuda, H. andKomamine, A. 1982. Lignin synthesis and its related enzymes as markers of trachearyelement differentiation in single cells isolated from the mesophyll ofZinnia elegans. Planta155: 423–430.CrossRefGoogle Scholar
  7. Goldberg, R., Lë, T. andCatesson, A.-M. 1985. Localization and properties of cell wall enzyme activities related to the final stages of lignin biosynthesis. J. Exp. Bot.36: 503–510.Google Scholar
  8. Harkin, J.M. andObst, J.R. 1973. Lignification in trees: Induction of exclusive peroxidase participation. Science180: 296–298.Google Scholar
  9. Hepler, P.K., Rice, R.M. andTerranova, W.A. 1972. Cytochemical localization of peroxidase activity in wound vessel members ofColeus. Can. J. Bot.50: 977–983.Google Scholar
  10. Imberty, A., Goldberg, R. andCatesson, A.-M. 1985. Isolation and characterization ofPopulus isoperoxidases involved in the last step of lignification. Planta164: 221–226.CrossRefGoogle Scholar
  11. Laemmli, U.K. 1970. Cleavage of structural proteins during assembly of the head bacteriophage T4. Nature227: 680–685.CrossRefPubMedGoogle Scholar
  12. Lagrimini, L.M., Burkhart, W., Moyer, M. andRothstein, S. 1987. Molecular cloning of complementary DNA encoding the lignin-forming peroxidase from tobacco: Molecular analysis and tissue-specific expression. Proc. Natl. Acad. Sci. USA84: 7542–7546.Google Scholar
  13. Mäder, M., Nessel, A. andBopp, M. 1977. Über die physiologische Bedeutung der Peroxidase-Isoenzymgruppen des Tabaks anhand einiger biochemischer Eigenschaften II. pH-Optima, Michaelis-Konstanten, Maximale Oxidationsraten. Z. Pflanzenphysiol.82: 247–260.Google Scholar
  14. Mäder, M., Ungemach, J. andSchlo, P. 1980. The role of peroxidase isoenzyme groups ofNicotiana tabacum in hydrogen peroxide formation. Planta147: 467–470.CrossRefGoogle Scholar
  15. Masuda, H., Fukuda, H. andKomamine, A. 1983. Changes in peroxidase isoenzyme patterns during tracheary element differentiation in a culture of single cells isolated from the mesophyll ofZinnia elegans. Z. Pflanzenphysiol.112: 417–426.Google Scholar
  16. Mazza, G. andWelinder, K.G. 1980. Covalent structure of turnip peroxidase 7. Cyanogen bromide fragments, complete structure and comparison to horseradish peroxidase C. Eur. J. Biochem.108: 481–489.PubMedGoogle Scholar
  17. McDougall, G.J. 1991. Cell-wall-associated peroxidases and lignification during growth of flax fibres. J. Plant Physiol.139: 182–186.Google Scholar
  18. Pedreño, M.A., Ros Barceló, A., Sabater, F. andMuñoz, R. 1989. Control by pH of cell wall peroxidase activity involved in lignification. Plant Cell Physiol.30: 237–241.Google Scholar
  19. Sato, Y., Sugiyama, M., Górecki, R.J., Fukuda, H. andKomamine, A. 1993. Interrelationship between lignin deposition and the activities of peroxidase isoenzymes in differentiating tracheary elements ofZinnia: analysis using L-α-aminooxy-β-phenyl-propionic acid and 2-aminoindan-2-phosphonic acid. Planta189: 584–589.CrossRefGoogle Scholar
  20. Sato, Y., Sugiyama, M., Komamine, A. andFukuda, H. 1995. Separation and characterization of the isoenzymes of wall-bound peroxidase from culturedZinnia cells during tracheary element differentiation. Planta196: 141–147.CrossRefGoogle Scholar
  21. Smith, C.G., Rodgers, M.W., Zimmerlin, A., Ferdinando, D. andBolwell, G.P. 1994. Tissue and subcellular immunolocalization of enzymes of lignin synthesis in differentiating and wounded hypocotyl tissue of French bean (Phaseolus vulgaris L.). Planta192: 155–164.Google Scholar
  22. Sugiyama, M., Fukuda, H. andKomamine, A. 1986. Effects of nutrient limitation and gamma irradiation on tracheary element differentiation and cell division in single mesophyll cells ofZinnia elegans. Plant Cell Physiol.27: 601–606.Google Scholar
  23. van Huystee, R.B. 1987. Some molecular aspects of plant peroxidase. Biosynthetic studies. Annu. Rev. Plant Physiol.38: 205–219.Google Scholar
  24. Welinder, K.G. 1979. Amino acid sequence studies of horseradish peroxidase. Amino acid carboxyl terminal, cyanogen bromide and tryptic fragments, the complete sequence, and some structural characteristics of horseradish peroxidase C. Eur. J. Biochem.96: 483–502.PubMedGoogle Scholar

Copyright information

© The Botanical Society of Japan 1995

Authors and Affiliations

  • Yasushi Sato
    • 1
  • Munetaka Sugiyama
    • 1
  • Takashi Takagi
    • 1
  • Hiroo Fukuda
    • 1
  1. 1.Biological Institute, Faculty of ScienceTohoku UniversitySendaiJapan
  2. 2.Department of Biology, Faculty of ScienceEhime UniversityMatsuyamaJapan

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