1887

Abstract

SUMMARY: A biochemical and histochemical investigation with specific substrates and inhibitors was used to visualize protease, esterase and aminopeptidase activities produced during penetration of and cuticles by hyphae of the entomopathogenic fungus Two endoproteases, and aminopeptidase and esterase activities, were mainly localized in simple and complex appressoria and germinating conidia. The effect of inhibitors on two characterized proteases (Pr1 and Pr2) and aminopeptidase activity in appressorial plates was quantified by microdensitometric measurement of reaction products. Pr1 and Pr2 activities were differentially inhibited by various protease inhibitors. Pr1, Pr2, esterase, aminopeptidase and -acetylglucosaminidase (exochitinase) activities were present during penetration as detected directly following desorption from fungal and cuticle components. The proteases produced were fractionated, and were shown by immunological and enzymological criteria to be the same as those produced in culture media. The sequence of enzyme appearance showed that production of proteolytic enzymes precedes exochitinase production. No production of endochitinase was found before or during hyphal penetration of the cuticle.

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/content/journal/micro/10.1099/00221287-133-5-1371
1987-05-01
2021-10-20
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References

  1. Bell R. A., Joachim F. G. 1976; Techniques for rearing laboratory colonies of tobacco hornworms and pink bollworms. Annals of the Entomological Society of America 69:365–375
    [Google Scholar]
  2. Brockerhoff H., Jensen R. G. 1974 Lipolytic Enzymes New York & London: Academic Press;
    [Google Scholar]
  3. Chang P. L. Y., Trevithick V. R. 1974; How important is secretion of enzymes through apical cell walls of fungi?. Archives of Microbiology 101:281–293
    [Google Scholar]
  4. Charnley A. K. 1984; Physiological aspects of destructive pathogenesis in insects by fungi: a speculative review. In Invertebrate-Microbial Interactions pp. 229–270 British Mycological Society Symposium 6 Anderson J. M., Rayner A. D. M., Walton D. W. H. Edited by Cambridge: Cambridge University Press;
    [Google Scholar]
  5. Fargues J. 1984; Adhesion of the fungal spore to the insect cuticle in relation to pathogenicity. In Infection Processes of Fungi pp. 90–111 Roberts D. W., Aist J. R. Edited by New York: The Rockefeller Foundation;
    [Google Scholar]
  6. Gabriel B. P. 1968; Histochemical study of the insect cuticle infected by the fungus Entomophthora coronala. Journal of Invertebrate Pathology 11:82–89
    [Google Scholar]
  7. Gomori G. 1945; The microtechnical demonstration of sites of lipase activity. Proceedings of the Society for Experimental Biology and Medicine 58:362–364
    [Google Scholar]
  8. Gull K., Trinci A. P. J. 1974; Detection of areas of wall differentiation in fungi using fluorescent staining. Archives of Microbiology 96:53–57
    [Google Scholar]
  9. Hackman R. H. 1980; Biochemical methods (proteins). In Cuticle Techniques in Arthropods pp. 145–184 Miller T. A. Edited by New York: SpringerVerlag;
    [Google Scholar]
  10. Luppa H., Andra J. 1982; The histochemistry of carboxylester hydrolases: problems and possibilities. Histochemical Journal 15:111–137
    [Google Scholar]
  11. Mirelman D., Galun E., Sharon N., Lotan R. 1975; Inhibition of fungal growth by wheat germ agglutinin. Nature; London: 256414–416
    [Google Scholar]
  12. Moore S., Stein W. H. 1948; Photometric ninhydrin method for use in the chromatography of amino acids. Journal of Biological Chemistry 76:367–388
    [Google Scholar]
  13. Neville A. C. 1975 Biology of the Arthropod Cuticle Berlin: Springer-Verlag;
    [Google Scholar]
  14. Pearse A. G. E. 1972 Histochemistry, Theoretical and Applied 2, 3rd edn.. Edinburgh & London: Churchill Livingston;
    [Google Scholar]
  15. Ramakrishnan L., Staples R. C. 1970; Changes in ribonucleic acids during uredospore differentiation. Phytopathology 60:1087–1091
    [Google Scholar]
  16. Ratault C., Vey A. 1977; Production d’esterases et de N-acetyl-β-d-glucosaminidase dans le tegument du Coleoptere Oryctes rhinoceros par le champignon entomopathogene Metarhizium anisopliae. Entomo-phaga 22:289–294
    [Google Scholar]
  17. Smith L. R., Gilkerson E. 1979; Quantitation of glycosaminoglycan hexosamine using 3-methyl-2-benzothiazolone hydrazone hydrochloride. Analytical Biochemistry 98:478–480
    [Google Scholar]
  18. Staples R. C., Hoch H. C. 1984; A sensing mechanism in rust uredospore germlings responsive to host morphology starts the cell cycle. In Infection Processes of Fungi pp. 126–136 Roberts D. W., Aist J. R. Edited by USA: The Rockefeller Foundation;
    [Google Scholar]
  19. Starkey P. M., Barrett A. J. 1976; Human lysosomal elastase, catalytic and immunological properties. Biochemical Journal 155:265–271
    [Google Scholar]
  20. Stevenson G. T. 1974 Immunisation with antigen coupled to an immunosorbent Nature; London: 247477–478
    [Google Scholar]
  21. St Leger R. J., Charnley A. K., Cooper R. M. 1986a; Cuticle degrading enzymes of entomo-pathogenic fungi. Synthesis in culture on cuticle. Journal of Invertebrate Pathology 48:85–95
    [Google Scholar]
  22. St Leger R. J., Cooper R. M., Charnley A. K. 1986b; Cuticle degrading enzymes of entomo-pathogenic fungi: cuticle degradation in vitro by enzymes from entomopathogens. Journal of Invertebrate Pathology 47:167–177
    [Google Scholar]
  23. St Leger R. J., Cooper R. M., Charnley A. K. 1986C; Cuticle-degrading enzymes of entomo-pathogenic fungi : regulation of production of chitin-olytic enzymes. Journal of General Microbiology 132:1509–1517
    [Google Scholar]
  24. St Leger R. J., Charnley A. K., Cooper R. M. 1986d; Cuticle-degrading enzymes of entomo-pathogenic fungi : mechanisms of interaction between pathogen enzymes and insect cuticle. Journal of Invertebrate Pathology 47:295–302
    [Google Scholar]
  25. St Leger R. J., Charnley A. K., Cooper R. M. 1987; Characterization of cuticle-degrading proteases produced by the entomopathogen Metarhizium anisopliae. Archives of Biochemistry and Biophysics 253:221–232
    [Google Scholar]
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