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Separation and characterization of the isoenzymes of wall-bound peroxidase from cultured Zinnia cells during tracheary element differentiation

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Abstract

Cell wall-bound peroxidase (EC 1.11.1.7) isoenzymes (P1-P5) from cells of Zinnia elegans L. that were differentiating into tracheary elements were separated and characterized to obtain information about the relationships between these isoenzymes and the biosynthesis of lignin. Fractionation of Zinnia cells by centrifugation in solutions of Percoll revealed that P1, P2, and P5 were present in differentiated tracheary elements. These peroxidase isoenzymes were separated by several column-chromatographic steps. During hydrophobic chromatography on Phenyl Superose, P5 activity was separated into activities P5A and P5B. Enzymatically pure preparations of P1, P3, P5A, and P5B were finally obtained and used for the characterization of each isoenzyme. The optimum pH was 5.5–6.0 for P1, 5.0–7.5 for P3, 5.0 for P5A, and 4.0 for P5B. Each of the isoenzymes oxidized coniferyl alcohol efficiently, whereas p-coumaryl alcohol and sinapyl alcohol were poor substrates for all the isoenzymes. An absolute requirement for Ca2+ ions was demonstrated for P3. Based on these results, possible roles of peroxidase isoenzymes in the formation of lignin during the differentiation of tracheary elements are discussed.

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Abbreviations

DAB:

diaminobenzidine

GTA:

equal proportions of 3,3-dimethylglutaric acid, tris(hydroxymethyl)aminomethane, and 2-amino-2-methyl-1,3-propanediol

TE:

tracheary element

References

  • Abeles, F.B., Biles, C.L. (1991) Characterization of peroxidases in lignifying peach fruit endocarp. Plant Physiol. 95, 269–273

    Google Scholar 

  • Bao, W., O'Malley, D.M., Whetten, R., Sederoff, R.R. (1993) A laccase associated with lignification in loblolly pine xylem. Science 260, 672–674

    Google Scholar 

  • Church, D.L., Galston, A.W. (1988) 4-Coumarate:coenzyme A ligase and isoperoxidase expression in Zinnia mesophyll cells induced to differentiate into tracheary elements. Plant Physiol. 88, 679–684

    Google Scholar 

  • Driouich, A., Laine, A.-C., Vian, B., Faye, L. (1992) Characterization and localization of lacasse forms in stem and cell cultures of sycamore. Plant J. 2, 13–24

    Google Scholar 

  • Fergus, B.J., Goring, D.A.I. (1970) The location of guaiacyl and syringyl lignins in birch xylem tissue. Holzforschung 24, 113–117

    Google Scholar 

  • Ferrer, M.A., Pedreño, M.A., Muñoz, R., Ros 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

    Google Scholar 

  • Fukuda, H. (1992) Tracheary element formation as a model system of cell differentiation. Int. Rev. Cytol. 136, 289–332

    Google Scholar 

  • Fukuda, H., Komamine, A. (1980) Establishment of an experimental system for the study of tracheary element differentiation from single cells isolated from the mesophyll of Zinnia elegans. Plant Physiol. 65, 61–64

    Google Scholar 

  • Fukuda, H., Komamine, A. (1982) Lignin synthesis and its related enzymes as markers of tracheary-element differentiation in single cells isolated from the mesophyll of Zinnia elegans. Plant Physiol. 65, 57–60

    Google Scholar 

  • Goldberg, R., Lê, T., Catesson, 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 

  • Hepler, P.K., Rice, R.M., Terranova, W.A. (1972) Cytochemical localization of peroxidase activity in wound vessel members of Coleus. Can. J. Bot. 50, 977–983

    Google Scholar 

  • Higuchi, T. (1985) Biosynthesis of lignin. In: Biosynthesis and biodegradation of wood components, pp. 141–160, Academic Press Inc., New York

    Google Scholar 

  • Imberty, A., Goldberg, R., Catesson, A.-M. (1985) Isolation and characterization of Populus isoperoxidases involved in the last step of lignification. Planta 164, 221–226

    Google Scholar 

  • Kohda, S. (1981) Kanshoueki no shurui to tyousei hou. In: Tanpakushitu kouso no kiso jikken hou, pp. 429–437, Horio, T., Yamashita, J., eds. Nankohdoh, Tokyo (in Japanese)

    Google Scholar 

  • Liu, L., Dean, J.F.D., Friedman, W.E., Eriksson, K.-E.L. (1994) A laccase-like phenoloxidase is correlated with lignin biosynthesis in Zinnia elegans stem tissues. Plant J. 6, 213–224

    Google Scholar 

  • Mäder, M., Nessel, A., Bopp, M. (1977) Über die physiologische Bedeutung der Peroxidase-Isoenzym-gruppen des Tabaks anhand einiger biochemischer Eigenschaften II. pH-Optima, Michaelis-Konstanten, Maximale Oxidationsraten. Z. Pflanzenphysiol. 82, 247–260

    Google Scholar 

  • Mäder, M., Ungemach, J., Schlo, P. (1980) The role of peroxidase isoenzyme groups of Nicotiana tabacum in hydrogen peroxide formation. Planta 147, 467–470

    Google Scholar 

  • Masuda, H., Fukuda, H., Komamine, A. (1983) Changes in peroxidase isoenzyme patterns during tracheary element differentiation in a culture of single cells isolated from the mesophyll of Zinnia elegans. Z. Pflanzenphysiol. 112, 417–426

    Google Scholar 

  • McDougall, G.J. (1991) Cell-wall-associated peroxidases and lignification during growth of flax fibres. J. Plant Physiol. 139, 182–186

    Google Scholar 

  • Pedreño, M.A., Ros Barceló, A., Sabater, F., Muñoz, R. (1989) Control by pH of cell wall peroxidase activity involved in lignification. Plant Cell Physiol. 30, 237–241

    Google Scholar 

  • Sato, Y., Sugiyama, M., Gorecki, R.J., Fukuda, H., Komamine, A. (1993) Interrelationship between lignin deposition and the activities of peroxidase isoenzymes in differentiating tracheary elements of Zinnia: analysis using L-α-aminooxy-β-phenylpropionic acid and 2-aminoindan-2-phosphonic acid. Planta 189, 584–589

    Google Scholar 

  • Smith, C.G., Rodgers, M.W., Zimmerlin, A., Ferdinando, D., Bolwell, G.P. (1994) Tissue and subcellular immunolocalization of enzymes of lignin synthesis in differentiating and wounded hypocotyl tissue of French bean (Phaseolus vulgaris L.). Planta 192, 155–164

    Google Scholar 

  • Sterjiades, R., Dean, J.F.D., Eriksson, K.-E.L. (1992) Laccase from sycamore maple (Acer pseudoplatanus) polymerizes monolignols. Plant Physiol. 99, 1162–1168

    Google Scholar 

  • Sterjiades, R., Dean, J.F.D., Gamble, G., Himmelsbach, D.S., Eriksson, K.-E.L. (1993) Extracellular laccases and peroxidases from sycamore maple (Acer psudoplatanus) cell-suspension cultures. Reactions with monolignols and lignin model compounds. Planta 190, 75–87

    Google Scholar 

  • Sticher, L., Penel, C., Greppin, H. (1981) Calcium requirement for the secretion of peroxidase by plant cell suspension. J. Cell Sci. 48, 345–355

    Google Scholar 

  • Sugiyama, M., Fukuda, H., Komamine, A. (1986) Effects of nutrient limitation and gamma irradiation on tracheary element differentiation and cell division in single mesophyll cells of Zinnia elegans. Plant Cell Physiol. 27, 601–606

    Google Scholar 

  • van Huystee, R.B. (1987) Some molecular aspects of plant peroxidase. Biosynthetic studies. Annu. Rev. Plant Physiol. 38, 205–219

    Google Scholar 

  • Welinder, K.G. (1985) Plant peroxidases. Their primary, secondary and tertiary structures, and relation to cytochrome c peroxidase. Eur. J. Biochem. 151, 497–504

    Google Scholar 

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The authors are very grateful to Professor M. Tanahashi of Gifu University for providing hydroxycinnamyl alcohols. This work was supported in part by Grants-in-Aid from the Ministry of Education, Science and Culture of Japan to H.F.

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Sato, Y., Sugiyama, M., Komamine, A. et al. Separation and characterization of the isoenzymes of wall-bound peroxidase from cultured Zinnia cells during tracheary element differentiation. Planta 196, 141–147 (1995). https://doi.org/10.1007/BF00193227

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  • DOI: https://doi.org/10.1007/BF00193227

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