Summary
A cytochemical technique for the ultrastructural localization of chitin in tomato root cells infected byFusarium oxysporum f. sp.radicis-lycopersici is reported. Chitinase was complexed to colloidal gold and thin sections were incubated with the enzyme-gold complex. This technique yielded a more uniform distribution of gold particles over the fungus wall, compared to that obtained with the lectin-gold technique. Both techniques revealed no labelling of the fungus cytoplasm, except for organelles resembling Woronin bodies. No significant labelling of either healthy or infected root cells was seen except for the secondary walls of vessels and, occasionally, that of adjoining parenchyma cells. The importance of this technique in studying the development of the pathogen within host cells is discussed.
Similar content being viewed by others
References
Arroyo-Begovich, A. &Carabez-Trejo, A. (1982) Location of chitin in the cyst wall ofEntamoeba invadens with colloidal gold tracers.J. Parasit. 68, 253–8.
Bendayan, M. (1981a) Ultrastructural localization of nucleic acids by the use of enzyme-gold complexes.J. Histochem. Cytochem. 29, 531–41.
Bendayan, M. (1981b) Electron microscopical localization of nucleic acids by means of nuclease-gold complexes.Histochem. J. 13, 699–710.
Bracker, C. E. (1967) Ultrastructure of fungi.A. Rev. Phytopath. 5, 343–74.
Charest, P. M., Miville, L. &Pauzé, F. J. (1983) Note sur une technique simple pour l'obtention de plantules stériles et son application pour des études en histopathologie.Phytoprotection 64, 31–4.
Charest, P. M., Ouellette, G. B. &Pauzé, F. J. (1984). Cytological observations of early infection process byFusarium oxysporum f. sp.radicis-lycopersici in tomato plants.Can. J. Bot. 62, 1232–44.
Farkas, V. (1979) Biosynthesis of cell wall of fungi.Microbiol. Rev. 43, 117–44.
Frens, G. (1973) Controlled nucleation for regulation of the particle size in monodisperse gold suspensions.Nature phys. Sci. 241, 20–2.
Goodman, S. L., Hodges, G. M. &Livingston, D. C. (1980) A review of the colloidal gold marker system.Scanning Electron Microscopy II, 133–46.
Hadwiger, L. A., Beckman, J. M. &Adams, M. J. (1981) Localization of fungal components in the pea-Fusarium interaction detected immunochemically with anti-chitosan and anti-fungal cell wall antisera.Plant Physiol. 67, 170–5.
Horisberger, M. &Rosset, J. (1977a) Colloidal gold, a useful marker for transmission and scanning electron microscopy.J. Histochem. Cytochem. 25, 295–305.
Horisberger, M. &Rosset, J. (1977b) Localization of wheat germ agglutinin receptor sites on yeast cells by scanning electron microscopy.Experentia 32, 998–1000.
Horisberger, M. &Vonlanthen, M. (1977) Location of mannan and chitin on thin sections of budding yeasts with gold markers.Arch. Mikrobiol. 115, 1–7.
Molano, J., Bowers, B. &Cabib, E. (1980) Distribution of chitin in the yeast cell wall.J. Cell Biol. 85, 199–212.
Pusztai, A. (1964) Hexosamines in the seeds of higher plants (Spermatophytes).Nature, Lond. 201, 1328–9.
Ride, J. P. &Drysdale, R. B. (1971) A chemical method for estimatingFusarium oxysporum f. lycopersici in infected tomato plants.Physiol. Plant Path. 1, 409–20.
Tronchin, G., Poulain, D., Herbaut, J. &Biguet, J. (1981) Localization of chitin in the cell wall ofCandida albicans by means of wheat germ agglutinin fluorescence and ultrastructural studies.Eur. J. Cell Biol. 26, 121–8.
Vian, B., Brillouet, J. M. &Satiat-Jeunemaitre, B. (1983) Ultrastructural visualization of xylans in cell walls of hardwood by means of xylanase-gold complex.Biol. Cell. 49, 179–82.
Wergin, W. P. (1973) Development of Woronin bodies from microbodies inFusarium oxysporum f. sp.lycopersici.Protoplasma 76, 249–60.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Chamberland, H., Charest, P.M., Ouellette, G.B. et al. Chitinase-gold complex used to localize chitin ultrastructurally in tomato root cells infected byFusarium oxysporum f. sp.radicislycopersici, compared with a chitin specific gold-conjugated lectin. Histochem J 17, 313–321 (1985). https://doi.org/10.1007/BF01004593
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01004593