1887

Abstract

The conidial germling of produces an appressorium upon contact with a hard hydrophobic surface. We have conducted an investigation into how this entomopathogen mediates intracellularly the inductive signal to shift from polarized germ-tube growth to non-polarized appressorial growth. During sporulation, conidia accumulated Ca but there was no evidence for a gradient of Ca in the spore which could establish the initial polarity. Calmodulin, however, was localized at the poles of the conidia, near the site of germ-tube emergence. Exposing conidia to Ca deprivation or calmodulin antagonists inhibited germination and polar growth. Disruption of Ca gradients by ionophoresis did not prevent germination but caused the multiple emergence of branched germ-tubes from conidia. These findings indicate that Ca plays a fundamental role in establishing the dominance of apical growth. Although an external source of Ca is not required for appressorium formation, germlings producing appressoria took up Ca when available. The Ca accumulated in the cell wall, plasma membrane and organelles suggesting that these may function as Ca stores for Ca-stimulated exocytosis of cell-wall materials. Mitochondria and vacuoles sequestered Ca indicating that they play a role in maintaining low cytoplasmic concentrations of Ca consistent with the reorganization of the cytoskeleton required for appressorial growth. Several Ca-binding proteins in appressoria may provide an energy-independent component of Ca buffering in the cytoplasm. The results indicate that the apical Ca gradient is disrupted during differentiation and subsequent differential Ca redistribution in the cell enlargement zone coincides with germ-tube swelling. cAMP may also be involved by potentiating the effects of small changes in Ca concentration and stimulating exocytosis of mucus components required for adhesion, which is a prerequisite for differentiation.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-9-1779
1990-09-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/9/mic-136-9-1779.html?itemId=/content/journal/micro/10.1099/00221287-136-9-1779&mimeType=html&fmt=ahah

References

  1. Bowman E. J., Bowman B. J. 1988; Purification of vacuolar membrane, mitochondria and plasma membranes from Neurosporacrassaand modes of discriminating among the different H+- ATPases. Methods in Enzymology 157:562–573
    [Google Scholar]
  2. 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]
  3. Butt T. M., Hoch H. C., Staples R. C., Stleger R. J. 1989; Use of fluorochromes in the study of fungal cytology and differentiation. Experimental Mycology 13:303–320
    [Google Scholar]
  4. Caswell A. H. 1979; Methods of measuring intracellular calcium. International Review of Cytology 56:145–181
    [Google Scholar]
  5. Cornelius G., Nakashima H. 1987; Vacuoles play a decisive role in calcium homeostasis in Neurospora crassa. Journal of General Microbiology 133:2341–2347
    [Google Scholar]
  6. Deber C. M., Tom-KUN J., Mack E., Grinstein S. 1985; Bromo- A23187: a nonfluorescent calcium ionophore for use with fluorescent probes. Analytical Biochemistry 146:349–352
    [Google Scholar]
  7. Demarty M., Morvan C., Thellier M. 1984; Calcium and the cell wall. Plant, Cell and Environment 7:441–448
    [Google Scholar]
  8. Gerisch G. 1987; Cyclic AMP and other signals controlling cell development and differentiation in Dictyostelium. Annual Review of Biochemistry 56:853–879
    [Google Scholar]
  9. Goettel M. S., Stleger R. J., Rizzo N. W., Staples R. C., Roberts D. W. 1989; Ultrastructural localization of a cuticledegrading protease produced by the entomopathogenic fungus Metarhiziumanisopliaeduring penetration of host (Manducasexta) cuticle. Journal of General Microbiology 134:2233–2239
    [Google Scholar]
  10. Goettel M. S., Stleger R. J., Bhairi S., Jung M. K., Oakley B. R., Roberts D. W., Staples R. C. 1990; Transformation of the entomopathogenic fungus Metarhiziumanisopliaeusing the Aspergillusnidulans ben A3gene. Current Genetics 17:129–132
    [Google Scholar]
  11. Hanson B. A. 1989; The effects of lithium on the growth, morphology and biochemistry of Neurospora crassa. Program of the XV Fungal Genetics Conference,University of California, Los Angeles
    [Google Scholar]
  12. Hauber I., Herth W., Reiss H. D. 1984; Calmodulin in tipgrowing cells, visualized by fluorescing calmodulin-binding pheno- thiazines. Planta 162:33–39
    [Google Scholar]
  13. Hidaka H., Tanaka T. 1987; TransmembraneCa2+signalling and a new class of inhibitors. Methods in Enzymology 139:570–582
    [Google Scholar]
  14. Jackson S. L., Heath I. B. 1989; Effects of exogenous calcium ions on tip growth, intracellular Ca2+ concentration and actin arrays in hyphae of the fungus Saprolegniaferax. Experimental Mycology 13:1–12
    [Google Scholar]
  15. Maresca B., Kobayashi G. S. 1989; Dimorphism in Histoplasmacapsulatum: a model for the study of cell differentiation in pathogenic fungi. Microbiological Reviews 53:186–209
    [Google Scholar]
  16. Maruyama K., Mikawa T., Ebashi S. 1984; Detection of calcium binding proteins by 45Ca2+ autoradiography on nitrocellulose membrane after sodium dodecyl sulfate gel electrophoresis. Journal of Biochemistry 95:511–519
    [Google Scholar]
  17. Mckerracher L. J., Heath I. B. 1987; Cytoplasmic migration and intracellular organelle movements during tip growth of fungal hyphae. Experimental Mycology 11:79–100
    [Google Scholar]
  18. Picton J. M., Steer M. W. 1982; A model for the mechanism of tip extension in pollen tubes. Journal of Theoretical Biology 98:15–20
    [Google Scholar]
  19. Picton J. M., Steer M. W. 1983; Evidence for the role of Ca2+ ions in tip extension in pollen tubes. Protoplasma 115:11–17
    [Google Scholar]
  20. Picton J. M., Steer M. W. 1985; The effects of ruthenium red, lanthanum, fluorescein isothiocyanate and trifluoperazine on vesicle transport, vesicle fusion and tip extension in pollen tubes. Planta 163:20–26
    [Google Scholar]
  21. Reissig J. L., Kinney S. G. 1983; Calcium as a branching signal in Neurospora crassa. Journal of Bacteriology 154:1397–1402
    [Google Scholar]
  22. Rothermel J. D., Perillo N. L., Marks J. S., Parkerbotelho L. M. 1984; Effects of the specific cAMP antagonist (Rp)- Adenosine cyclic 3',5'-phosphorothioate, on the cAMP-dependent protein kinase-induced activity of hepatic glycogen phosphorylase and glycogen synthase. Journal of Biological Chemistry 259:15294–15300
    [Google Scholar]
  23. Schmid J., Harold F. M. 1988; Dual roles for calcium ions in apical growth of Neurospora crassa. Journal of General Microbiology 134:2623–2631
    [Google Scholar]
  24. Stleger R. J., Cooper R. M., Charnley A. K. 1987; Production of cuticle-degrading enzymes by the entomopathogenMetarhiziumanisopliaeduring infection of cuticles from Calliphoravomitoriaand Manducasexta. Journal of General Microbiology 133:1371–1382
    [Google Scholar]
  25. Stleger R. J., Butt T. M., Roberts D. W., Staples R. C. 1989; a.Production in vitroof appressoria by the entomopathogenic fungus Metarhiziumanisopliae. Experimental Mycology 13:274–288
    [Google Scholar]
  26. Stleger R. J., Butt T.M.STAPLES, Roberts D. W. 1989b; Synthesis of proteins including a cuticle degrading protease during differentiation of the entomopathogenic fungus Metarhiziumanisopliae. Experimental Mycology 13:253–262
    [Google Scholar]
  27. St leger R. J., roberts D. W., staples D. W. 1989c; Calcium and calmodulin-mediated protein synthesis and protein phosphorylation during germination, growth and protein production by Metarhiziumanisopliae. Journal of General Microbiology 135:2141–2154
    [Google Scholar]
  28. St leger R. J., roberts D. W. 1989d; Novel GTP-binding proteins in plasma membranes of the fungus Metarhi-ziumanisopliae.Biochemical and Biophysical Research Communications. Journal of General Microbiology 164:362–366
    [Google Scholar]
  29. St leger R. J., roberts D. W., staples R. C. 1989a; A model to explain differentiation of appressoria by germlings of Metarhiziumanisopliae.Journal of Invertebrate Pathology(in the Press).
    [Google Scholar]
  30. St leger R. J., Laccetti L. B., Staples R. C., Roberts D. W. 1990b; Protein kinases in the entomopathogenic fungus Metarhi-ziumanisopliae. Journal of General Microbiology 136:1401–1411
    [Google Scholar]
  31. Takeuchi Y., Schmid J., Caldwell J. H., Harold T. M. 1988; Transcellular ion currents and extension of Neurosporacrassahyphae. Journal of Membrane Biology 101:33–41
    [Google Scholar]
  32. Uchida E., Ohsumi Y., Anraka Y. 1988; Purification of yeast vacuolar membrane H+-ATPase and enzymological discrimination of three ATP-driven proton pumps in Saccharomyces cerevisiae. Methods in Enzymology 157:544–562
    [Google Scholar]
  33. Vanlookerencampagne M. M., Grneux C., Vaneijk R. 1988; Two dephosphorylation pathways of inositol 1,4,5-triphosphate in Dictyosteliumdiscoideumhomogenates. Biochemical Journal 254:343–350
    [Google Scholar]
  34. Wessels J.G.H. 1986; Cell wall synthesis in apical hyphal growth. International Review of Cytology 104:37–79
    [Google Scholar]
  35. Zacharuk R. Y. 1970; Fine structure of the fungus Metarhiziumanisopliaeinfecting three species of larval elateridae (coleoptera). II. Conidial germ tubes and appressoria. Journal of Invertebrate Pathology 15:81–91
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-9-1779
Loading
/content/journal/micro/10.1099/00221287-136-9-1779
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error