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Planta

, Volume 157, Issue 1, pp 22–31 | Cite as

Chitinase in bean leaves: induction by ethylene, purification, properties, and possible function

  • T. Boller
  • A. Gehri
  • F. Mauch
  • U. Vögeli
Article

Abstract

Ethylene induced an endochitinase in primary leaves of Phaseolus vulgaris L. The enzyme formed chitobiose and higher chitin oligosaccharides from insoluble, colloidal or regenerated chitin. Less than 5% of the total chitinolytic activity was detected in an exochitinase assay proposed by Abeles et al. (1970, Plant Physiol. 47, 129–134) for ethylene-induced chitinase. In ethylene-treated plants, chitinase activity started to increase after a lag of 6 h and was induced 30 fold within 24 h. Exogenously supplied ethylene at 1 nl ml−1 was sufficient for half-maximal induction, and enhancement of the endogenous ethylene formation also enhanced chitinase activity. Cycloheximide prevented the induction. Among various hydrolases tested, only chitinase and, to a lesser extent, β-1,3-glucanase were induced by ethylene. Induction of chitinase by ethylene occurred in many different plant species. Ethylene-induced chitinase was purified by affinity chromatography on a column of regenerated chitin. Its apparent molecular weight obtained by sodium dodecyl sulfate-gel electrophoresis was 30,000; the molecular weight determined from filtration through Sephadex G-75 was 22,000. The purified enzyme attacked chitin in isolated cell walls of Fusarium solani. It also acted as a lysozyme when incubated with Micrococcus lysodeikticus. It is concluded that ethylene-induced chitinase functions as a defense enzyme against fungal and bacterial invaders.

Key words

Chitinase Defense (against bacteria, fungi) Enzyme induction Ethylene Lysozyme Phaseolus (chitinase) 

Abbreviations

ACC

1-aminocyclopropane-1-carboxylic acid

AVG

aminoethoxyvinylglycine

GlcNAc

N-acetylglucos-amine

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References

  1. Abeles, F.B. (1973) Ethylene in plant biology. Academic Press, New York LondonGoogle Scholar
  2. Abeles, F.B., Bosshart, R.P., Forrence, L.E., Habig, W.H. (1970) Preparation and purification of glucanase and chitinase from bean leaves. Plant Physiol. 47, 129–134Google Scholar
  3. Altmann, A., Kaur-Sawhney, R., Galston, A.W. (1977) Stabilization of oat leaf protoplasts through polyamine-mediated inhibition of senescence. Plant Physiol. 60, 570–574Google Scholar
  4. Araki, Y., Ito, E. (1975) A pathway for chitosan formation in Mucor rouxii. Enzymatic deacetylation of chitin. Eur. J. Biochem. 55, 71–78Google Scholar
  5. Berger, L.R., Reynolds, D.M. (1958) The chitinase system of a strain of Streptomyces griseus. Biochim. Biophys. Acta 29, 522–534Google Scholar
  6. Boller, T., Kende, H. (1979) Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol. 63, 1123–1132Google Scholar
  7. Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254Google Scholar
  8. Cabib, E., Bowers, B. (1971) Chitin and yeast budding. J. Biol. Chem. 246, 152–159Google Scholar
  9. Eastwell, K.C., Bassi, P.K., Spencer, M.E. (1978) Comparison and evaluation of methods for the removal of ethylene and other hydrocarbons from air for biological studies. Plant Physiol. 62, 723–726Google Scholar
  10. Glazer, A.N., Barel, A.O., Howard, J.B., Brown, D.M. (1969) Isolation and characterization of fig lysozyme. J. Biol. Chem. 244, 3583–3589Google Scholar
  11. Hadwiger, L.A., Beckman, J.M. (1980) Chitosan as a component of pea-Fusarium solani interactions. Plant Physiol. 66, 205–211Google Scholar
  12. Hestrin, H., Feingold, E.S., Schramm, M. (1955) Hexoside hydrolases. Methods Enzymol. 1, 231–257Google Scholar
  13. Howard, J.B., Glazer, A.N. (1967) Studies of the physicochemical and enzymatic properties of papaya lysozyme. J. Biol. Chem. 242, 5715–5723Google Scholar
  14. Konze, J.R., Kwiatowski, M.K. (1981) Rapidly induced ethylene formation after wounding is controlled by the regulation of ACC synthesis. Planta 151, 327–330Google Scholar
  15. Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227, 680–685Google Scholar
  16. Lieberman, M. (1979) Biosynthesis and action of ethylene. Annu. Rev. Plant Physiol. 30, 533–591Google Scholar
  17. Molano, J., Duran, A., Cabib, E. (1977) A rapid and sensitive assay for chitinase using tritiated chitin. Anal. Biochem. 83, 648–656Google Scholar
  18. Molano, J., Polacheck, I., Duran, A., Cabib, E. (1979) An endochitinase from wheat germ. J. Biol. Chem. 254, 4901–4907Google Scholar
  19. Montalbini, P., Elstner, E.F. (1977) Ethylene evolution by rustinfected, detached bean (Phaseolus vulgaris L.) leaves susceptible and hypersensitive to Uromyces phaseoli (Pers.) Wint. Planta 135, 301–306Google Scholar
  20. Nichols, J.E., Beckman, J.M., Hadwiger, L.A. (1980) Glycosidic enzyme activity in pea tissue and pea-Fusarium solani interactions. Plant Physiol. 66, 199–204Google Scholar
  21. Otakara, A. (1961) Studies on the chitinolytic enzymes in blackkoji mold. II. Purification of chitinase. Agr. Biol. Chem. 25, 54–60Google Scholar
  22. Pegg, G.F. (1976) The involvement of ethylene in plant pathogenesis. In: Encyclopedia of plant physiology, N. S., vol. 4, pp. 582–591, Heitefuss, R., Williams, P.H., eds. Springer, Berlin Heidelberg New YorkGoogle Scholar
  23. Pegg, G.F. (1977) Glucanohydrolases of higher plants: a possible defense mechanism against parasitic fungi. In: Cell wall biochemistry related to specificity in host-plant pathogen interactions, pp. 305–345, Solheim, B., Raa, J., eds. Universitetsforlaget, Tromsø Oslo BergenGoogle Scholar
  24. Pegg, G.F., Vessey, J.C. (1973) Chitinase activity in Lycopersicon esculentum and its relationship to the in vivo lysis of Verticillium albo-atrum mycelium. Physiol. Plant Pathol. 3, 371–382Google Scholar
  25. Pegg, G.F., Young, D.H. (1981) Changes in glycosidase activity and their relationship to fungal colonisation during infection of tomato by Verticillium albo-atrum. Physiol. Plant Pathol. 19, 371–382Google Scholar
  26. Powning, R.F., Irzykiewicz, H. (1965) Studies on the chitinase system in bean and other seeds. Comp. Biochem. Physiol. 14, 127–133Google Scholar
  27. Reissig, J.L., Strominger, J.L., Leloir, L.F. (1955) A modified colorimetric method for the estimation of N-acetylamino sugars. J. Biol. Chem. 217, 959–966Google Scholar
  28. Rupley, J.A. (1964) The hydrolysis of chitin by concentrated hydrochloric acid, and the preparation of low-molecular-weight substrates for lysozyme. Biochim. Biophys. Acta 83, 245–255Google Scholar
  29. Wargo, P.M. (1975) Lysis of the cell wall of Armillaria mellea by enzymes from forest trees. Physiol. Plant Pathol. 5, 99–105Google Scholar
  30. Yang, S.F., Pratt, H.K. (1978) The physiology of ethylene in wounded plant tissues. In: Biochemistry of wounded plant tissues, pp. 595–622, Kahl, G., ed. Walter de Gruyter, BerlinGoogle Scholar

Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • T. Boller
    • 1
  • A. Gehri
    • 1
  • F. Mauch
    • 1
  • U. Vögeli
    • 1
  1. 1.Botanisches Institut der UniversitätBaselSwitzerland

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