Accumulation of pathogenesis-related proteins in the epidermis of tomato leaves infected by Cladosporium fulvum

  • J. P. Wubben
  • C. A. Eijkelboom
  • P. J. G. M. De Wit


Upon infection byCladosporium fulvum, tomato plants start to produce pathogenesis-related (PR) proteins. The PR proteins 1,3-β-glucanase, chitinase, and PR-1b accumulated near the stomata in the lower epidermis ofC. fulvum-inoculated tomato leaves as could be determined by immunolocalization with polyclonal antibodies. However, no differences in accumulation of PR proteins between a compatible and an incompatible interaction were found. Results obtained from enzyme activity measurements of 1,3-β-glucanase and chitinase on similar leaf material as used for the immunolocalization did not fully reflect the immunolocalization data. The antibodies possibly detect only the extracellular but not the intracellular enzymes. The accumulation of PR proteins near the stomata might be part of a general defence response of plants against pathogens and potential pathogens.

Additional keywords

1,3-β-glucanase chitinase PR-1b stoma immumolocalization 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Broglie, K., Chet, I., Holliday, M., Cressman, R., Biddle, P., Knowlton, S., Mauvais, C.J. & Broglie, R., 1991. Transgenic plants with enhanced resistance to the fungal pathogenRhizoctonia solani. Science 254: 1194–1197.PubMedCrossRefGoogle Scholar
  2. De Wit, P.J.G.M., 1977. A light and scanning-electron microscopic study of infection of tomato plants by virulent and avirulent races ofCladosporium fulvum. Netherlands Journal of Plant Pathology 83: 109–122.CrossRefGoogle Scholar
  3. De Wit, P.J.G.M. & Flach, W., 1979. Differential accumulation of phytoalexins in tomato leaves, but not in fruits after inoculation with virulent and avirulent races ofCladosporium fulvum. Physiological Plant Pathology 15: 257–267.CrossRefGoogle Scholar
  4. De Wit, P.J.G.M., Buurlage, M.B. & Hammond, K.E., 1986. The occurrence of host-, pathogen- and interaction- specific proteins in the apoplast ofCladosporium fulvum (syn.Fulvia fulva)-infected tomato leaves. Physiological and Molecular Plant Pathology 29: 159–172.Google Scholar
  5. Farmer, E.E. & Ryan, C.A., 1990. Interplant communication: Airborne methyl jasmonate induces synthesis of proteinase inhibitors in plant leaves. Proceedings of the national Academy of Sciences, USA 87: 7713–7716.CrossRefGoogle Scholar
  6. Joosten, M.H.A.J. & De Wit, P.J.G.M., 1988. Isolation, purification and preliminary characterization of a protein specific for compatibleCladosporium fulvum (syn.Fulvia fulva)-tomato interactions. Physiological and Molecular Plant Pathology 33: 241–253.CrossRefGoogle Scholar
  7. Joosten, M.H.A.J. & De Wit, P.J.G.M., 1989. Identification of several pathogenesis-related proteins in tomato leaves inoculated withCladosporium fulvum (syn.Fulvia fulva) as 1,3-β-glucanases and chitinases. Plant Physiology 89: 945–951.PubMedCrossRefGoogle Scholar
  8. Joosten, M.H.A.J., Bergmans, C.J.B., Meulenhoff, E.J.S., Cornelissen, B.J.C. & De Wit, P.J.G.M., 1990. Purification and serological characterization of three basic 15-kilodalton pathogenesisrelated proteins from tomato. Plant Physiology 94: 585–591.PubMedCrossRefGoogle Scholar
  9. Kauffmann, S., Legrand, M., Geoffroy, P. & Fritig, B., 1987. Biological function of ‘pathogenesis-related’ proteins: four PR proteins of tobacco have 1,3-β-glucanase activity. The EMBO Journal 6: 3209–3212.PubMedGoogle Scholar
  10. Kombrink, E., Beerhues, L., Garcia-Garcia, F., Hahlbrock, K., Müller, M., Schröder, M., Witte, B. & Schmelzer, E., 1993. Expression patterns of defense-related genes in infected and uninfected plants. In: Fritig, B. & Legrand, M. (Eds), Developments in plant pathology, Vol. 2, Mechanisms of plant defense responses. Kluwer Academic Publishers, Dordrecht, the Netherlands. p. 236–249.Google Scholar
  11. Lawton, K., Uknes, S., Friedrich, L., Gaffney, T., Alexander, D., Goodman, R., Metraux, J.P., Kessmann, H., Ahl Goy, P., Gut Rella, M., Ward, E. & Ryals, J., 1993. The molecular biology of systemic acquired resistance. In: Fritig, B. & Legrand, M. (Eds), Developments in plantpathology, Vol. 2, Mechanisms of plant defense responses. Kluwer Academic Publishers, Dordrecht, the Netherlands. p. 422–432.Google Scholar
  12. Legrand, M., Kauffmann, S., Geoffroy, P. & Fritig, B., 1987. Biological function of pathogenesis-related proteins: Four tobacco pathogenesis-related proteins are chitinases. Proceedings of the National Academy of Sciences, USA 84: 6750–6754.CrossRefGoogle Scholar
  13. Linthorst, H.J.M., 1991. Pathogenesis-related proteins of plants. Critical Reviews in Plant Sciences 10: 123–150.CrossRefGoogle Scholar
  14. Mauch, F., Mauch-Mani, B. & Boller, T., 1988. Antifungal hydrolases in pea tissue. II. Inhibition of fungal growth by combinations of chitinase and β-1,3-glucanase. Plant Physiology 88: 936–942.PubMedCrossRefGoogle Scholar
  15. Mauch, F., Meehl, J.B. & Staehelin, L.A., 1992. Ethylene-induced chitinase and β-1,3-glucanase accumulate specifically in the lower epidermis and along the vascular strands of bean leaves. Planta 186: 367–375.CrossRefGoogle Scholar
  16. Melchers, L.S., Sela-Buurlage, M.B., Vloemans, S.A., Woloshuk, C.P., Van Roekel, J.S.C., Pen, J., Van den Elzen, P.J.M. & Cornelissen, B.J.C.., 1993. Extracellular targeting of the vacuolar tobacco proteins AP24, chitinase and β-1,3-glucanase in transgenic plants. Plant Molecular Biology 21: 583–593.PubMedCrossRefGoogle Scholar
  17. Neuhaus, J.-M., Ahl-Goy, P., Hinz, U., Flores, S. & Meins, F.Jr., 1991a. High-level expression of a tobacco chitinase gene inNicotiana sylvestris. Susceptibility of transgenic plants toCercospora nicotianae infection. Plant Molecular Biology 16: 141–151.PubMedCrossRefGoogle Scholar
  18. Neuhaus, J.-M., Sticher, L., Meins, F.Jr. & Boller, T., 1991b. A short C-terminal sequence is necessary and sufficient for the targeting of chitinases to the plant vacuole. Proceedings of the National Academy of Sciences, USA 88: 10362–10366.CrossRefGoogle Scholar
  19. Ohashi, Y. & Matsuoka, M., 1987. Localization of pathogenesis-related proteins in the epidermis and intercellular spaces of tobacco leaves after their induction by potassium salicylate or tobacco mosaic virus infection. Plant Cell Physiology 28: 1227–1235.Google Scholar
  20. Schlumbaum, A., Mauch, F., Vögeli, U. & Boller, T., 1986. Plant chitinases are potential inhibitors of fungal growth. Nature 324: 265–267.CrossRefGoogle Scholar
  21. Sela-Buurlage, M.B., Ponstein, A.S., Bres-Vloemans, S.A., Melchers, L.S., van den Elzen, P.J.M. & Cornelissen, B.J.C., 1993. Only specific tobacco (Nicotiana tabacum) chitinases and β-1,3-glucanases exhibit antifungal activity. Plant Physiology 101: 857–863.PubMedGoogle Scholar
  22. Van Lent, J.W.M. & Verduin, B.J.M., 1986. Detection of viral protein and particles in thin sections of infected plant tissue using immunogold labelling. In: Jones, R.A.C. & Torrance, L. (Eds), Developments in applied biology. 1. Developments and applications in virus testing. Wellesbourne, UK: 193–211.Google Scholar
  23. Vigers, A.J., Roberts, W.K. & Selitrennikoff, C.P., 1991. A new family of plant antifungal proteins. Molecular-Plant Microbe Interactions 4: 315–323.PubMedGoogle Scholar
  24. Woloshuk, C.P., Meulenhoff, J.S., Sela-Buurlage, M., Van den Elzen, P.J.M. & Cornelissen, B.J.C., 1991. Pathogen-induced proteins with inhibitory activity towardPhytophthora infestans. The Plant Cell 3: 619–628.PubMedCrossRefGoogle Scholar
  25. Wubben, J.P., Joosten, M.H.A.J., Van Kan, J.A.L. & De Wit, P.J.G.M., 1992. Subcellular localization of plant chitinases and 1,3-β-glucanases inCladosporium fulvum (syn.Fulvia fulva)-infected tomato leaves. Physiological and Molecular Plant Pathology 41: 23–32.CrossRefGoogle Scholar

Copyright information

© Koninklijke Nederlandse Planteziektenkundige Vereniging 1993

Authors and Affiliations

  • J. P. Wubben
    • 1
  • C. A. Eijkelboom
    • 1
  • P. J. G. M. De Wit
    • 1
  1. 1.Department of PhytopathologyWageningen Agricultural UniversityWageningenthe Netherlands

Personalised recommendations