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Cytochemical Labeling for Fungal and Host Components in Plant Tissues Inoculated with Fungal Wilt Pathogens

Published online by Cambridge University Press:  01 August 2004

G.B. Ouellette
Affiliation:
Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre, 1055 du P.E.P.S., P.O. Box 3800, Sainte-Foy, Quebec G1V 4C7, Canada
R.P. Baayen
Affiliation:
Plant Protection Service, P.O. Box 9102, NL-6700 HC Wageningen, The Netherlands
H. Chamberland
Affiliation:
Département de phytologie, Pavillon C.-E.–Marchand, Université Laval, Quebec G1K 7P4, Canada
M. Simard
Affiliation:
Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre, 1055 du P.E.P.S., P.O. Box 3800, Sainte-Foy, Quebec G1V 4C7, Canada
D. Rioux
Affiliation:
Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre, 1055 du P.E.P.S., P.O. Box 3800, Sainte-Foy, Quebec G1V 4C7, Canada
P.M. Charest
Affiliation:
Département de phytologie, Pavillon C.-E.–Marchand, Université Laval, Quebec G1K 7P4, Canada
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Abstract

Antibodies to detect pectin in present investigations attached to distinct fibrils in vessel lumina. In carnation infected with an isolate of Fusarium oxysporum f.sp., labeling of pathogen cells also occurred; in a resistant cultivar (cv.), it was coincident with proximate pectin fibrils and linked to altered fungal walls, which was the opposite in the susceptible cv., indicating that hindrance of pathogen ability to degrade pectin may be related to resistance. Labeling of the fungus in culture was nil, except in media containing pectin, showing that pectin is not native to the pathogen. Labeling of fungal walls for cellulose in elm (inoculated with Ophiostoma novo-ulmi) and carnation also occurred, linked to adsorbed host wall components. The chitin probe often attached to dispersed matter, in vessel lumina, traceable to irregularly labeled fungal cells and host wall degradation products. With an anti-horseradish peroxidase probe, host and fungal walls were equally labeled, and with a glucosidase, differences of labeling between these walls were observed, depending on pH of the test solution. Fungal extracellular matter and filamentous structures, present in fungal walls, predominantly in another elm isolate (Phaeotheca dimorphospora), did not label with any of the probes used. However, in cultures of this fungus, extracellular material labeled, even at a distance from the colony margin, with an anti-fimbriae probe.

Type
Biological Applications
Copyright
© 2004 Microscopy Society of America

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References

REFERENCES

Aist, J.R. (1983). Structural responses as resistance mechanisms. In The Dynamics of Host Defence, Bailey, J.A. & Deverall, B.J. (Eds.), pp. 3370. Sydney, Australia: Academic Press.
Albersheim, P. & Anderson, A.J. (1971). Proteins from plant cell walls inhibit polygalacturonases secreted by plant pathogens. Proc Natl Acad Sci USA 68, 18151819.Google Scholar
Albersheim, P., Jones, T.M., & English, P.D. (1969). Biochemistry of the cell wall in relation to infective processes. Annu Rev Phytopathol 7, 171194.Google Scholar
Baayen, R.P., O'Donnell, K., Bonants, P.J.M., Cigelnik, E., Kroon, L.P.N.M., Roebroeck, E.J.A., & Waalwijk, C. (2000). Gene genealogies and AFLP analyses in the Fusarium oxysporum complex identify monophyletic and nonmonophyletic formae speciales causing wilt and rot disease. Phytopathology 90, 891900.Google Scholar
Baayen, R.P., Ouellette, G.B., & Rioux, D. (1996). Compartmentalization of decay in carnations resistant to Fusarium oxysporum f.sp. dianthi. Phytopathology 86, 10181031.Google Scholar
Baayen, R.P., Schoffelmeer, E.A.M., Toet, S., & Elgersma, D.M. (1997). Fungal polygalacturonase activity reflects susceptibility of carnation cultivars to fusarium wilt. Eur J Plant Pathol 103, 1523.Google Scholar
Beckman, C.H. (1987). The Nature of Wilt Diseases of Plants. American Phytopathological Society, St. Paul, MN: APS Press.
Bendayan, M. (1989). Protein A-gold and protein G-gold postembedding immunoelectron microscopy. In Colloidal Gold: Principles, Methods, and Applications, Hayat, M.A. (Ed.), pp. 3495. New York: Academic Press.
Benhamou, N. & Asselin, A. (1989). Attempted localization of a substrate for chitinases in plant cells reveals abundant N-acetyl-D-glucosamine residues in secondary walls. Biol Cell 67, 341350.Google Scholar
Bonsen, K.J.M. & Kucera, L.J. (1990). Vessel occlusions in plants: Morphological, functional and evolutionary aspects. IAWA Bull n.s. 11, 393399.Google Scholar
Bonsen, K.J.M., Sheffer, R.J., & Elgersma, D.M. (1985). Barrier zone formation as a resistance mechanism of elms to Dutch elm disease. IAWA Bull 6, 7177.Google Scholar
Celerin, M., Ray, J.M., Schisler, N.J., Day, A.W., Stetler-Stevenson, W.G., & Laudenbach, D.E. (1996). Fungal fimbriae are composed of collagen. EMBO J 15, 44454453.Google Scholar
Chamberland, H. (1994). Gold labeling methods for the ultrastructural localization of host wall and pathogen components. In Host Wall Alterations by Parasitic Fungi, Petrini, O. & Ouellette, G.B. (Eds.), pp. 111. American Phytopathological Society, St. Paul, MN: APS Press.
Chamberland, H., Benhamou, N., Ouellette, G.B., & Pauzé, F.J. (1989). Cytochemical detection of saccharide residues in paramural bodies formed in tomato root cells infected by Fusarium oxysporum f.sp. radicis-lycopersici. Physiol Mol Plant Pathol 34, 131146.Google Scholar
Chamberland, H., Charest, P.M., Ouellette, G.B., & Pauzé, F.J. (1985). Chitinase-gold complex used to localize chitin ultrastructurally in tomato root cells infected by Fusarium oxysporum f.sp. radicis-lycopersici, compared with a chitin specific gold-conjugated lectin. Histochem J 17, 313321.Google Scholar
Chamberland, H., Ouellette, G.B., Pauzé, F.J., & Charest, P.M. (1991). Immunocytochemical localization of tomato pectinesterase in root cells of tomato plants infected by Fusarium oxysporum f.sp. radicis-lycopersici. Can J Bot 69, 12651274.Google Scholar
Charest, P.M., Ouellette, G.B., & Pauzé, F.J. (1984). Cytological observations of early infection process by Fusarium oxysporum f.sp. radicis-lycopersici in tomato plants. Can J Bot 62, 12321244.Google Scholar
Day, A.W., Gardiner, R.B., Smith, R., Svircev, A.M., & McKeen, W.E. (1986). Detection of fungal fimbriae by protein A-gold immunocytochemical labelling in host plants infected with Ustilago heufleri or Peronospora hyoscyami f.sp. tabacina. Can J Microbiol 32, 577584.Google Scholar
DesRochers, P. & Ouellette, G.B. (1994). Phaeotheca dimorphospora sp.nov.: Description et caractéristiques culturales. Can J Bot 72, 808817.Google Scholar
Dimond, A.E. (1970). Biophysics and biochemistry of the vascular wilt syndrome. Annu Rev Phytopathol 8, 301322.Google Scholar
Duchesne, L.C., Jeng, R.S., & Hubbes, M. (1985). Accumulation of phytoalexins in Ulmus americana in response to infection by a nonaggressive and an aggressive strain of Ophiostoma ulmi. Can J Bot 63, 678680.Google Scholar
Hutchison, K.A., Green, J.R., Wharton, P.S., & O'Connell, R.J. (2002). Identification and localisation of glycoproteins in the extracellular matrices around germ-tubes and appressoria of Colletotrichum species. Mycol Res 106, 729736.Google Scholar
Knox, J.P., Linstead, P.J., King, J., Cooper, C., & Roberts, K. (1990). Pectin esterification is spatially regulated both within cell walls and between developing tissues of root apices. Planta 181, 512521.Google Scholar
Mace, M.E. (1978). Contributions of tyloses and terpenoid aldehyde phytoalexins to Verticillium wilt resistance in cotton. Physiol Plant Pathol 12, 111.Google Scholar
Nicole, M., Ruel, K., & Ouellette, G.B. (1994). Fine morphology of fungal structures involved in host wall alteration. In Host Wall Alterations by Parasitic Fungi, Petrini, O. & Ouellette, G.B. (Eds.), pp. 1330. American Phytopathological Society, St. Paul, MN: APS Press.
O'Donnell, K., Kistler, H.C., Cigelnik, E., & Ploetz, R.C. (1998). Multiple evolutionary origins of the fungus causing Panama disease of banana: Concordant evidence from nuclear and mitochondrial gene genealogies. Proc Natl Acad Sci USA 95, 20442049.Google Scholar
Ouellette, G.B. (1980). Occurrence of tyloses and their ultrastructural differentiation from similarly configured structures in American elm infected by Ceratocystis ulmi. Can J Bot 58, 10561073.Google Scholar
Ouellette, G.B. & Baayen, R.P. (2000). Peculiar structures occurring in vessel walls of the susceptible carnation cultivar Early Sam infected with Fusarium oxysporum f.sp. dianthi. Can J Bot 78, 270277.Google Scholar
Ouellette, G.B., Baayen, R.P., Chamberland, H., Simard, M., & Charest, P.M. (2002a). Cytochemical localization of fungal wall components in host-pathogen interactions: Particular labeling with gold-complexed probes. Microsc Microanal 8 (Suppl. 2), 254255.Google Scholar
Ouellette, G.B., Baayen, R.P., Simard, M., & Rioux, D. (1999a). Ultrastructural and cytochemical study of colonization of xylem vessel elements of susceptible and resistant Dianthus caryophyllus by Fusarium oxysporum f.sp. dianthi. Can J Bot 77, 644663.Google Scholar
Ouellette, G.B., Baayen, R.P., Simard, M., & Rioux, D. (2002b). Reactions of paratracheal cells of resistant and susceptible carnation (Dianthus caryophyllus) cultivars to vascular invasion by Fusarium oxysporum f.sp. dianthi. New Phytol 156, 113128.Google Scholar
Ouellette, G.B., Chamberland, H., Goulet, A., Lachapelle, M., & Lafontaine, J.-G. (1999b). Fine structure of the extracellular sheath and cell walls in Ophiostoma novo-ulmi growing on various substrates. Can J Microbiol 45, 582597.Google Scholar
Ouellette, G.B., Méthot, N., Chamberland, H., Côté, C., & Lafontaine, J.-G. (1995). Cytology of irregular growth forms of Ophiostoma ulmi and Ophiostoma novo-ulmi growing through millipore membranes and sterilized elm wood sections. Can J Microbiol 41, 10951110.Google Scholar
Ouellette, G.B. & Rioux, D. (1992). Anatomical and physiological aspects of resistance to Dutch elm disease. In Defense Mechanisms of Woody Plants Against Fungi, Blanchette, R.A. & Biggs, A.R. (Eds.), pp. 257307. Berlin: Springer-Verlag.
Ouellette, G.B. & Rioux, D. (1993). Alterations of vessel elements and reactions of surrounding tissues in the DED syndrome. In Dutch Elm Disease Research: Cellular and Molecular Approaches, Sticklen, M.B. & Sherald, J.L. (Eds.), pp. 255292. New York: Springer-Verlag.
Ouellette, G.B., Rioux, D., Bussières, G., Simard, M., & Bernier, L. (2000). Extracellular sheath formation by Sphaeropsis hypodermia and association with its infection in elm trees. Phytoprotection 81, 6986.Google Scholar
Punja, Z.K. & Parker, M. (2000). Development of fusarium root and stem rot, a new disease on greenhouse cucumber in British Columbia, caused by Fusarium oxysporum f. sp. radicis-cucumerinum. Can J Plant Pathol 22, 349363.Google Scholar
Reynolds, E.S. (1963). The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. Cell Biol 17, 208212.Google Scholar
Rioux, D., Chamberland, H., Simard, M., & Ouellette, G.B. (1995). Suberized tyloses in trees: An ultrastructural and cytochemical study. Planta 196, 125140.Google Scholar
Rioux, D., Nicole, M., Simard, M., & Ouellette, G.B. (1998). Immunocytochemical evidence that secretion of pectin occurs during gel (gum) and tylosis formation in trees. Phytopathology 88, 494505.Google Scholar
Rioux, D. & Ouellette, G.B. (1991). Barrier zone formation in host and nonhost trees inoculated with Ophiostoma ulmi. I. Anatomy and histochemistry. Can J Bot 69, 20552073.Google Scholar
Shigo, A. & Tippett, J.T. (1981). Compartmentalization of American elm tissues infected by Ceratocystis ulmi. Plant Dis 65, 715718.Google Scholar
Tamaki, H. & Yamashina, S. (1994). Improved method for post-embedding cytochemistry using reduced osmium and LR White resin. J Histochem Cytochem 42, 12851293.Google Scholar
Tenberge, K.B., Beckedorf, M., Hoppe, B., Schouten, A., Solf, M., & von den Driesch, M. (2002). In situ localization of AOS in host–pathogen interactions. Microsc Microanal 8 (Suppl. 2), 250251.Google Scholar
Tenberge, K.B., Homann, V., Oeser, B., & Tudzynski, P. (1996). Structure and expression of two polygalacturonase genes of Claviceps pupurea oriented in tandem and cytological evidence for pectinolytic enzyme activity during infection of rye. Phytopathology 86, 10841097.Google Scholar