Surface polymers of the nematode-trapping fungus Free

Abstract

The nematophagous fungus captures nematodes using adhesive polymers present on special hyphae (traps) which form a three-dimensional network. To understand further the adhesion mechanisms, surface polymers were visualized by transmisson electron microscopy and characterized by chemical methods. Both traps and hyphae were surrounded by a fibrillar layer of extracellular polymers which stained with ruthenium red. The polymer layer was resistant to most of the chemicals and enzymes tested. However, part of the layer was removed by sonication in a Tris-buffer or by extraction in a chaotropic salt solution (LiCI), and the structure of the polymers was modified by treatment with Pronase E. Chemical analysis showed that the crude extracts of surface polymers removed by sonication or LiCl solution contained neutral sugars, uronic acids and proteins. Gel chromatography of the extracts revealed that the major carbohydrate-containing polymer(s) had a molecular mass of at least 100 kDa, containing neutral sugars (75% by weight, including glucose, mannose and galactose), uronic acids (6 %) and proteins (19 %). There was more polymer in mycelium containing trap-bearing cells than in vegetative hyphae. SDS-PAGE of the extracted polymers showed that the trap-forming cells contained at least one protein, with a molecular mass of approx. 32 kDa, not present on vegetative hyphae. Examining the capture of nematodes by traps of in which the layer of surface polymers was modified, or removed by chemical or enzymic treatments, showed that both proteins and carbohydrate surface polymers were involved in the adhesion process.

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1991-06-01
2024-03-29
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References

  1. Albersheim P., Nevins D. J., English P. D., Karr A. 1967; A method for the analysis of sugars in plant cell-wall polysaccharides by gas-liquid chromatography. Carbohydrate Research 5:340–345
    [Google Scholar]
  2. Beever R. E., Redgwell R. J., Dempsey G. P. 1979; Purification and chemical characterization of the rodlet layer of Neurospora crassa . Journal of Bacteriology 140:1063–1070
    [Google Scholar]
  3. Bitter T., Muir H. M. 1962; A modified uronic acid and carbazole reaction. Analytical Biochemistry 4:330–334
    [Google Scholar]
  4. Blakeney A. B., Harris P. J., Henry R. J., Stone B. A. 1983; A simple and rapid preparation of alditol acetates for monosaccharide analysis. Carbohydrate Research 113:291–299
    [Google Scholar]
  5. Borrebaeck C. A. K., Mattiasson B., Nordbring-Hertz B. 1984; Isolation and partial characterization of a carbohydrate-binding protein from a nematode-trapping fungus. Journal of Bacteriology 159:53–56
    [Google Scholar]
  6. Borrebaeck C. A. K., Mattiasson B., Nordbring-Hertz B. 1985; A fungal lectin and its apparent receptors on a nematode surface. FEMS Microbiology Letters 27:35–39
    [Google 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. Analytical Biochemistry 72:248–254
    [Google Scholar]
  8. Brooker B. E. 1979; Electron microscopy of the dextrans produced by lactic acid bacteria. In Microbial Polysaccharides and Polysac-charidases, Special Publication no. 3 of the Society for General Microbiology85–115 Berkeley R. C. W., Gooday G. W., Ellwood D. C. London: Academic Press;
    [Google Scholar]
  9. Catley B. J. 1988; Isolation and analysis of cell walls. In Yeast: a Practical Approach163–183 Campbell I., Duffus J. H. Oxford: IRL Press;
    [Google Scholar]
  10. Chaffin W. L., Stocco D. M. 1983; Cell wall proteins of Candida albicans . Canadian Journal of Microbiology 29:1438–1444
    [Google Scholar]
  11. Chattaway F. W., Shenolikar S., Barlow A. J. E. 1974; The release of acid phosphatase and polysaccharide- and protein- containing components from the surface of the dimorphic-forms of Candida albicans by treatment with dithiothreitol. Journal of General Microbiology 83:423–425
    [Google Scholar]
  12. Claverie-Martin F., Diaz-Torres M. R., Geoghegan M. J. 1986; Chemical composition and electron microscopy of the rodlet layer of Aspergillus nidulans conidia. Current Microbiology 14:221–225
    [Google Scholar]
  13. Cooper J. B., Varner J. E. 1983; Insolubilization of hydroxypro-line-rich cell wall glycoprotein in aerated carrot slices. Biochemical and Biophysical Research Communications 112:161–167
    [Google Scholar]
  14. Costerton J. W., Irvin R. T., Cheng K.-J. 1981; The role of bacterial surface structures in pathogenesis. CRC Critical Reviews in Microbiology 8:303–338
    [Google Scholar]
  15. Dubois M., Gilles K. A., Hamilton J. K., Rebers P. A., Smith F. 1956; Colorimetric method for determination of sugars and related substances. Analytical Chemistry 28:350–356
    [Google Scholar]
  16. Elson L. A., Morgan W. T. J. 1933; A colorimetric method for the determination of glucosamine and chondrosamine. Biochemical Journal 27:1824–1828
    [Google Scholar]
  17. Foerder C. A., Shapiro B. M. 1977; Release of ovoperoxidase from sea urchin eggs hardens the fertilization membrane with tyrosine crosslinks. Proceedings of the National Academy of Sciences of the United States of America 744214–4218
    [Google Scholar]
  18. Freifelder D. 1982; Hydrodynamic shear and sonic degradation. In Physical Biochemistry. Applications to Biochemistry and Molecular Biology685–688 San Francisco: W. H. Freeman and Company;
    [Google Scholar]
  19. Friman E., Olsson S., Nordbring-Hertz B. 1985; Heavy trap-formation by Arthrobotrys oligospora in liquid culture. FEMS Microbiology Ecology 31:17–21
    [Google Scholar]
  20. Gold R. E., Mendgen K. 1984; Cytology of basidiospore germination, penetration, and early colonization of Phaseolus vulgaris by Uromyces appendiculatus var. appendiculatus . Canadian Journal of Botany 62:1989–2002
    [Google Scholar]
  21. Gubler F., Hardham A. R. 1988; Secretion of adhesive material during encystment of Phytophthora cinnamomi zoospores, characterized by immunogold labelling with monoclonal antibodies to components of peripheral vesicles. Journal of Cell Science 90:225–235
    [Google Scholar]
  22. Hamer J. E., Howard R. J., Chumley F. G., Valent B. 1988; A mechanism for surface attachment in spores of a plant pathogenic fungus. Science 239:288–290
    [Google Scholar]
  23. Handley P. S., Hargreaves J., Harty D. W. S. 1988; Ruthenium red staining reveals surface fibrils and a layer external to the cell wall in Streptococcus salivarius HB and adhesion deficient mutants. Journal of General Microbiology 134:3165–3172
    [Google Scholar]
  24. Hyde K. D., Jones E. B. G. 1989; Observation on ascospore morphology in marine fungi and their attachment to surfaces. Botanica Marina 32:205–218
    [Google Scholar]
  25. Kennedy M. J. 1990; Models for studying the role of fungal attachment in colonization and pathogenesis. Mycopathologia 109:123–137
    [Google Scholar]
  26. Latge J.-P., Cole G. T., Horisberger M., Prevost M.-C. 1986; Ultrastructure and chemical composition of the ballastospore wall of Conidiobolus obscurus . Experimental Mycology 10:99–113
    [Google Scholar]
  27. Luft J. H. 1971; Ruthenium red and violet I. Chemistry, purification, methods of use for electron microscopy and mechanism of action. Anatomical Record 171:347–368
    [Google Scholar]
  28. Marrie T. J., Costerton J. W. 1981; The ultrastructure of Candida albicans infections. Canadian Journal of Microbiology 27:1156–1164
    [Google Scholar]
  29. Mims C. W., Richardson E. A. 1989; Ultrastructure of appressorium development by basidiospore germlings of the rust fungus Gymnosporangium juniperi-virginianae . Protoplasma 148:111–119
    [Google Scholar]
  30. Nicholson R. L., Epstein L. 1991; Adhesion of fungi to the plant surface: prerequisite for pathogenesis. In The Fungal Spore and Disease Initiation in Plants and Animals Cole C. T., Hoch H. C. New York: Plenum Press;
    [Google Scholar]
  31. Nicholson R. L., Moraes W. B. C. 1980; Survival of Colletotrichum graminicola: importance of the spore matrix. Phytopathology 70:255–261
    [Google Scholar]
  32. Nicholson R. L., Butler L. G., Asquith T. N. 1986; Glycoproteins from Colletotrichum graminicola that bind phenols: implications for survival and virulence of phytopathogenic fungi. Phytopathology 76:1315–1318
    [Google Scholar]
  33. Nicholson R. L., Yoshioka H., Yamaoka N., Kunoh H. 1988; Preparation of the infection court by Erysiphe graminis II. Release of esterase enzyme from conidia in relation to a contact stimulus. Experimental Mycology 12:336–349
    [Google Scholar]
  34. Nordbring-Hertz B., Mattiasson B. 1979; Action of a nematode-trapping fungus shows lectin-mediated host-microorganism interaction. Nature; London: 281477–479
    [Google Scholar]
  35. Nordbring-Hertz B., Veenhuis M., Harder W. 1984; Dialysis membrane technique for ultrastructural studies of microbial interactions. Applied and Environmental Microbiology 45:290–293
    [Google Scholar]
  36. Nordbring-Hertz B. 1988; Ecology and recognition in the nematode-nematophagous fungus system. In Advances in Microbial Ecology 1081–114 Marshall K. C. New York: Plenum Press;
    [Google Scholar]
  37. Nordbring-Hertz B., Mattiasson B. 1979; Action of a nematode-trapping fungus shows lectin-mediated host-microorganism interaction. Nature; London: 281477–479
    [Google Scholar]
  38. Nordbring-Hertz B., Veenhuis M., Harder W. 1984; Dialysis membrane technique for ultrastructural studies of microbial interactions. Applied and Environmental Microbiology 45:290–293
    [Google Scholar]
  39. Onyile A. B., Edwards H. H., Gessner R. V. 1982; Adhesive material of the hyphopodia of Buergenerula spartinae . Mycologia 74:777–784
    [Google Scholar]
  40. Peberdy J. F. 1985; Mycolytic enzymes. In Fungal Protoplasts. Applications in Biochemistry and Genetics31–44 Peberdy J. F., Ferenczy L. New York: Marcel Dekker, Inc;
    [Google Scholar]
  41. Ponton J., Jones J. M. 1986; Analysis of cell wall extracts of Candida albicans by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot techniques. Infection and Immunity 53:565–572
    [Google Scholar]
  42. Ramadoss C. S., Uhlig J., Carlson D. M., Butler L. G., Nicholson R. L. 1985; Composition of the mucilaginous spore matrix of Colletotrichum graminicola a pathogen of com, sorghum, and other grasses. Journal of Agricultural and Food Chemistry 33:728–732
    [Google Scholar]
  43. Rees D. A. 1972; Shapely polysaccharides. Biochemical Journal 126:257–273
    [Google Scholar]
  44. Spiro R. G. 1966; Analysis of sugars found in glycoproteins. Methods in Enzymology 8:3–25
    [Google Scholar]
  45. Sutherland I. W. 1989; Microbial polysaccharides - a comparison with eukaryotic polymers. In Mucus and Related Topics, Symposia of the Society for Experimental BiologyXLIII389–402 Chantler E., Ratcliffe N. A. Cambridge: The Company of Biologists Ltd;
    [Google Scholar]
  46. Taylor I. E. P, Cameron D. S. 1973; Preparation and quantitative analysis of fungal cell walls: strategy and tactics. Annual Review of Microbiology 27:243–259
    [Google Scholar]
  47. Teepe H., Bottge J.-A., Wostemeyer J. 1988; Isolation and electrophoretic analysis of surface proteins of the zygomycete Absidia glauca . FEBS Letters 234:100–106
    [Google Scholar]
  48. Tronchin G., Poulain D., Vernes A. 1984; Cytochemical and ultrastructural studies of Candida albicans III. Evidence for modification of the cell wall coat during adherence to human and buccal epithelial cells. Archives of Microbiology 139:221–224
    [Google Scholar]
  49. Veenhuis M., Nordbring-Hertz B., Harder W. 1985; An electron-microscopical analysis of capture and initial stages of penetration of nematodes by Arthrobotrys oligospora . Antonie van Leeuwenhoek 51:385–398
    [Google Scholar]
  50. Wessels J. G. H, Sietsma J. H. 1981; Fungal cell walls: a survey. In Plant Carbohydrates II: Extracellular Carbohydrates352–394 Tanner W., Loewus F. A. Berlin: Springer Verlag;
    [Google Scholar]
  51. Wessels J. G. H., Kreger D. R., Marchant R., Regensburg B. A., DeVries O. M. H. 1972; Chemical and morphological characterization of the hyphal cell wall surface of the basidiomycete Schizophyllum commune . Biochimica et Biophysica Acta 273:346–358
    [Google Scholar]
  52. Wu-Yuan C. D., Hashimoto T. 1977; Architecture and chemistry of microconidial walls of Trichophyton mentagrophytes . Journal of Bacteriology 129:1584–1592
    [Google Scholar]
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