Skip to main content
SpringerLink
Log in

An examination of the peroxidases from Lupinus albus L. hypocotyls

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Peroxidases (EC 1.11.1.7) from hypocotyls of Lupinus albus L. cv. Rio Maior have been characterised using one- and two-dimensional, native electrophoretic techniques. Data are presented showing the complexity in charge and molecular size or shape of these peroxidases. We report the finding of a new acidic peroxidase and several new basic peroxidases in these hypocotyls, and of their stability to treatments considered to break ligand-induced variants and conformational variants derived from differences in polypeptide folding. Densitometric data demonstrate that these new peroxidases contribute up to 60‰ of the total peroxidase activity in hypocotyls. Studies of intercellular fluid, cell-wall and soluble fractions, with assays of purity were conducted in an attempt to define the subcellular locations of these additional peroxidases. The acidic form (pI 4.1) is greatly enriched in soluble fractions, three of the basic peroxidases (pIs 9.5, 9.7 and >9.7) are strongly associated to the cell wall, ad a minor, basic component (pI 9.7) is enriched in the intercellular fluid. Individual peroxidase activities with the substrates coniferyl alcohol, ferulic acid or indole acetic acid were compared by densitometric analysis of zymograms with those for guaiacol, and notable differences between these peroxidases in their capacity to oxidise indole acetic acid in vitro were identified. The possible functions of these peroxidases in vivo and their implications to current understanding of peroxidases in L. albus are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

APAGE:

anionic polyacrylamide gel electrophoresis

CA:

coniferyl alcohol

CPAGE:

cationic polyacrylamide gel electrophoresis

IEF:

isoelectric focusin

NEIEF:

non-equilibrated isoelectric focusing

2D:

two dimensional

pI:

isoelectric point

RCPAGE:

reversed current polyacrylamide gel electrophoresis

References

  • Barceló, A.R., Ferrer, M.A., Florenciano, E.G., Muñoz, R. (1991) The tonoplast localization of two basic isoperoxidases of high pI in Lupinus. Bot. Acta. 104, 272–278

    Google Scholar 

  • Barceló, A.R., Muñoz, R., Sabater, F. (1987) Lupin peroxidases. I. Isolation and characterization of cell wall bound isoperoxidase activity. Physiol. Plant. 71, 448–454

    Google Scholar 

  • Barceló, A.R., Muñoz, R., Sabater, F. (1989a) Substrate specificity of soluble lupin isoperoxidases. Biol. Plant. 31, 161–167

    Google Scholar 

  • Barceló, A.R., Muñoz, R., Sabater, F. (1989b) Subcellular location of basic and acid soluble isoperoxidases in Lupinus. Plant Science 63, 31–38

    Google Scholar 

  • Barceló, A.R., Pedreño, M.A., Ferrer, M.A., Sabater, F., Muñoz, R. (1990) Indole-3-methanol is the main product of the oxidation of indole-3-acetic acid catalyzed by two cytosolic basic isoperoxidases from Lupinus. Planta 181, 448–450

    Google Scholar 

  • Barceló, A.R., Pedreño, M.A., Muñoz, R., Sabater, F. (1988) Lupin peroxidases. II. Binding of acidic peroxidases to cell walls. Physiol. Plant. 73, 238–244

    Google Scholar 

  • Barceló, A.R., Pedreño, M.A., Muñoz, R., Sabater, F. (1989c) Physiological significance of the binding of the acidic peroxidases to cell walls of lupin. Physiol. Plant. 75, 267–274

    Google Scholar 

  • Cordewener, J., Booij, H., Van der Zandt, H., Engelen, F., Van Kammen, A., Vries, S. (1991) Tunicamycin inhibited carrot somatic embryogenesis can be restored by secreted cationic peroxidase isozymes. Planta 184, 478–486

    Google Scholar 

  • Douce, R., Bourguinin, J., Brouquisse, R., Neuburger, M. (1987) Isolation of plant mitochondria: Principles and criteria of integrety. Methods Enzymol. 148, 403–415

    Google Scholar 

  • Epstein, C.J., Shechter, A.N. (1968) An approach to the problem of conformational isoenzymes. Ann. N.Y. Acad. Sci. 151, 85–101

    Google Scholar 

  • Espelie, K.E., Franceschi, V.R., Kolattukudy, P.E. (1986) Immunocytochemical localization and time course of appearance of an anionic peroxidase associated with suberisation in wound-healing potato tuber tissue. Plant Physiol. 81, 487–492

    Google Scholar 

  • Ferrer, M.A., Pedreño, M.A., Caldrón, A.A., Muñoz, R., Barceló, A.R. (1990a) Distribution of isoflavones in lupin hypocotyls. Possible control of cell peroxidase activity involved in lignification. Physiol. Plant. 79, 610–616

    Google Scholar 

  • Ferrer, M.A., Pedreño, M.A., Muñoz, R., Barceló, A.R. (1990b) Oxidation of coniferyl alcohol by cell well peroxidases at the expense of indole-3-acetic acid and O2. FEBS Lett. 276, 127–130

    Google Scholar 

  • Ferrer, M.A., Pedreño, M.A., Muñoz, R., Barceló, A.R. (1991a) Soluble peroxidase gradients in lupin hypocotyls and the control of the level of polarly transported indole-3-acetic acid. J. Plant Growth Regul. 10, 139–146

    Google Scholar 

  • Ferrer, M.A., Pedreño, M.A., Muñoz, R., Barceló, A.R. (1991b) Integrated control by genistein of cell wall peroxidase activities involved in growth and differentiation. In: Biochemical, molecular and physiological aspects of plant peroxidases, pp. 353–358, Lobarzewski, J., Greppin, H., Penel, C., Gaspar, T., eds. University of Geneva, Switzerland

    Google Scholar 

  • Fry, S.C. (1979) Phenolic components of the primary cell wall and their possible role in the hormonal regulation of growth. Planta 146, 343–351

    Google Scholar 

  • Fry, S.C. (1982a) Phenolic components of primary cell wall. Biochem. J. 203, 493–504

    Google Scholar 

  • Fry, S.C. (1982b) Isodityrosine, a new cross-linking amino acid from plant cell wall glycoproteins. Biochem. J. 204, 449–455

    Google Scholar 

  • Fry, S.C. (1986) Cross-linking of matrix polymers in the growing cell walls of angiosperms. Annu. Rev. Plant Physiol. 37, 165–186

    Google Scholar 

  • Gaspar, T., Penel, C., Hagege, D., Greppin, H. (1991) Peroxidases in plant growth, differentiation and developmental processes. In: Biochemical, molecular and physiological aspects of plant peroxidases, pp. 249–280, Lobarzewski, J., Greppin, H., Penel, C., Gaspar, T., eds. University of Geneva, Switzerland

    Google Scholar 

  • Goldberg, R., Pang, A., Rolando, C., Franceschi, C., Catesson, A.M. (1991) Cell wall peroxidases and lignification: Tissue and substrate specificity. In: Biochemical, molecular and physiological aspects of plant peroxidases, pp. 209–220, Lobarzewski, J., Greppin, H., Penel, C., Gaspar, T., eds. University of Geneva, Switzerland

    Google Scholar 

  • Gove, J.P., Hoyle, M.C. (1975) The isozymic similarity of indole acetic acid oxidase to peroxidase in birch and horseradish. Plant Physiol. 56, 684–687

    Google Scholar 

  • Halliwell, B. (1978) Lignin synthesis: The generation of hydrogen peroxidase and Superoxide by horseradish peroxidase and its stimulation by mangenese (II) and phenols. Planta 140, 81–88

    Google Scholar 

  • Jackson, P., Ricardo, C.P.P. (1992) Cytochrome c aided resolution of Lupinus albus isoperoxidases in a cathodal polyacrylamide gel electrophoresis system. Anal. Biochem. 200, 36–41

    Google Scholar 

  • Joersbo, M., Andersen, J.M., Okkels, F.T., Rajagopal, R. (1989) Isoperoxidases as markers of somatic embryogenesis in carrot cell suspension cultures. Physiol. Plant. 76, 10–16

    Google Scholar 

  • Mäder, M. (1980) Origin of the heterogeneity of peroxidase isoenzymes group G1 from Nicotiana tabacum. I. Conformation. Z. Pflanzenphysiol. 96, 283–296

    Google Scholar 

  • Mato, M.C., Vietez, A.M. (1986) Changes in auxin protectors and IAA oxidases during the rooting of chestnut shoots in vitro. Physiol. Plant. 66, 491–494

    Google Scholar 

  • Mato, M.C., Rúa, M.L., Ferro, E. (1988) Changes in levels of peroxidases and phenolics during root formation in Vitis cultured in vitro. Physiol. Plant. 72, 84–88

    Google Scholar 

  • O'Farrell, P.Z., Goodman, H.M., O'Farrel, P.H. (1977) High resolution, two dimensional electrophoresis of basic as well as acid proteins. Cell 12, 1133–1142

    Google Scholar 

  • Rao, K.V., Suprasanna, P., Reddy, G.M. (1990) Biochemical changes in embryogenic and non-embryogenic calli of Zea mays L. Plant Sci. 66, 127–130

    Google Scholar 

  • Ros, A., Pedreño, M.A., Muñoz, R., Sabater, F. (1986) Specific binding and Ca++-mediated of acidic peroxidases to lupin cell walls and membrane fractions, respectively. Note on the subcellular localization of acidic isoperoxidases. In: Molecular and physiological aspects of plant peroxidase, pp. 165–168, Greppin, H., Penel, C., Gaspar, T., eds. University of Geneva, Switzerland

    Google Scholar 

  • Sánchez-Bravo, J., Ortuño, A., Acosta, M., Sabater, F. (1986) Distribution of indole-3-acetic acid in relation to the growth of etioliated Lupinus albus hypocotyls. Physiol. Plant. 66, 509–514

    Google Scholar 

  • Srivastava, O.P., Van Huystee, R.B. (1977) Interactions among phenols and peroxidase isozymes. Bot. Gaz. 138, 457–464

    Google Scholar 

  • Welinder, K.G. (1991) The plant peroxidase superfamily. In: Biochemical, molecular and physiological aspects of plant peroxidases, pp. 3–13, Lobarzewski, J., Greppin, H., Penel, C., Gaspar, T., eds. University of Geneva, Switzerland

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Química e Biológica, Instituto de Tecnologia, Apartado 127, 2780, Oeiras, Portugal

    Philip Jackson & Cândido P. P. Ricardo

  2. Departamento de Botânica e Engenharia Biológica, Instituto Superior de Agronomia, 1399, Lisboa Codex, Portugal

    Cândido P. P. Ricardo

Authors
  1. Philip Jackson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cândido P. P. Ricardo
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jackson, P., Ricardo, C.P.P. An examination of the peroxidases from Lupinus albus L. hypocotyls. Planta 194, 311–317 (1994). https://doi.org/10.1007/BF00197530

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00197530

Key words

Search

Navigation