Summary
Cell-wall regeneration and reversion of protoplasts ofSchizophyllum commune were investigated using electron microscopic methods and X-ray diffraction.
After 3 hours of regeneration protoplasts have formed a loosely organized wall which does not react with Thiéry's stain for periodic acid sensitive carbohydrates. This wall largely consists of chitin microfibrils which are adpressed to the plasmalemma and which are covered by loose aggregates of alkali-soluble S-glucan (α-1,3-glucan). Both components are microcrystalline, at least partly. Walls formed in the presence of polyoxin D only consist of thick loose fibers of S-glucan.
From 3 hours onward the inner chitin microfibrils of the wall of the primary cells become embedded in alkali-insoluble material that stains heavily with the Thiéry reagent and probably is similar to the R-glucan of the mature wall (i.e., β-1,3-β-1,6-glucan). The outer chitin microfibrils remain free of this matrix and are covered by S-glucan only.
Bud-like structures that arise have the same wall architecture as the primary cells,i.e., only the inner chitin microfibrils are embedded in R-glucan and the S-glucan forms a fluffy coat. The walls of hyphal tubes that arise are distinct, however, in that all chitin microfibrils are embedded in R-glucan and the S-glucan forms a compact coat.
Cytoplasmic vesicles are sparse in primary cells except at the sites of emergence of budlike structures and hyphae. They continue to be present in the apex of growing hyphae.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Bartnicki-Garcia, S., 1973: Fundamental aspects of hyphal morphogenesis. In: Microbial differentiation; Proc. 23th Symp. Soc. Gen. Microbiol. (London, 1973), pp. 245–267. Cambridge: University Press.
Carbonell, L. M., F. Gil, andF. Yegres, 1973: Cell wall regeneration of protoplasts ofHistoplasma capsulatum. In: Yeast, mould and plant protoplasts (J. R. Villanueva, I. García-Acha, S. Gascon, andF. Uruburu, eds.); Proc. Third Intern. Symp. Yeast Protoplasts (Salamanca, 1972), pp. 93–103. London and New York: Academic Press.
—,F. Kanetsuna, andF. Gil, 1970: Chemical morphology of glucan and chitin in the cell wall of the yeast phase ofParacoccidioides brasiliensis. J. Bacteriol.101, 636–642.
Cook, G. M. W., andR. W. Stoddart, 1973: Surface carbohydrates of the eukaryotic cell. London and New York: Academic Press.
Edwards, M. R., andR. W. Stevens, 1963: Fine structure ofListena monocytogenes. J. Bacteriol.86, 414–428.
Fultz, S. A., andR. A. Woolf, 1972: Surface structure inAllomyces during germination and growth. Mycologia64, 212–218.
Gabriel, M., 1970: Cell wall regeneration inRhizopus nigricans protoplasts. In: Yeast protoplasts (O. Nečas, andA. Svoboda, eds.); Proc. second intern, symp. yeast protoplasts (Brno, 1968), pp. 147–151. Brno: University J. E. Purkyně.
Girbardt, M., 1969: Die Ultrastruktur der Apikalregion von Pilzhyphen. Protoplasma67, 413–441.
Grove, S. N., andC. E. Bracker, 1970: Protoplasmic organization of hyphal tips among fungi: Vesicles and Spitzenkörper. J. Bacteriol.104, 989–1009.
—,K. B. Oujezdsky, andP. J. Szaniszlo, 1973: Budding in the dimorphic fungusPhialophora dermatitidis. J. Bacteriol.115, 323–329.
Heath, I. B., andA. D. Greenwood, 1970: The structure and formation of lomasomes. J. gen. Microbiol.62, 129–137.
Hegnauer, H., andH. R. Hohl, 1973: A structural comparison of cyst and germ tube wall inPhytophtbora palmivora. Protoplasma77, 151–163.
Hemmes, D. E., andH. R. Hohl, 1971: Ultrastructural aspects of encystation and cystgermination inPhytophtbora parasitica. J. Cell Sci.9, 175–192.
Koltin, Y., J. G. H. Wessels, andP. van der Valk, 1973: Chlamydospores ofSchizophyllum commune. Arch. Mikrobiol.91, 179–183.
Kreger-van Rij, N. J. W., andM. Veenhuis, 1971: A comparative study of the cell wall structure of basidiomycetous and related yeasts. J. gen. Microbiol.68, 87–95.
Marchant, R., andR. T. Moore, 1973: Lomasomes and plasmalemmasomes in fungi. Protoplasma76, 235–247.
—, andD. G. Smith, 1967: Wall structure and bud formation inRhodotorula glutinis. Arch. Mikrobiol.58, 248–256.
McCully, E. K., andC. E. Bracker, 1972: Apical vesicles in growing bud cells of heterobasidiomycetous yeasts. J. Bacteriol.109, 922–926.
Moor, H., andK. Mühlethaler, 1963: Fine structure in frozen-etched yeast cells. J. Cell Biol.17, 609–628.
Niederpruem, D. J., andJ. G. H. Wessels, 1969: Cytodifferentiation and morphogenesis inSchizopbyllum commune. Bacteriol. Rev.33, 505–535.
Pojnar, E., J. H. M. Willison, andE. C. Cocking, 1967: Cell-wall regeneration by isolated tomato-fruit protoplasts. Protoplasma64, 460–480.
Raudaskoski, M., 1970: Occurrence of microtubules and microfilaments, and origin of septa in dikaryotic hyphae ofSchizophyllum commune. Protoplasma70, 415–422.
Reese, E. T., andM. Mandels, 1966: β-Glucanases other than cellulase. In: Methods in Enzymology (S. P. Colowick, andN. O. Kaplan, eds.), Vol.8, 4th edition, pp. 607–615. New York: Academic Press.
Reynolds, E. S., 1963: The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell Biol.17, 208–212.
Roland, J.-C. etB. Vian, 1971: Réactivité du plasmalemme vegetal. Etude cytochimique. Protoplasma73, 121–137.
San-Blas, G., andL. M. Carbonell, 1974: Chemical and ultrastructural studies on the cell walls of the yeast-like and mycelial forms ofHistoplasma farciminosum. J. Bacteriol.119, 602–611.
Sietsma, J. H., J. J. Child, L. R. Nesbitt, andR. H. Haskins, 1975: Ultrastructural aspects of wall regeneration byPythium protoplasts. Antonie van Leeuwenhoek41, 17–23.
Strunk, C., 1969: Licht- und elektronenmikroskopische Untersuchungen an regenerierenden Protoplasten vonPolystictus versicolor. Z. allg. Mikrobiol.9, 205–216.
—, 1970: Protoplasts of moulds-The present state of knowledge. In: Yeast protoplasts (O. Nečas, andA. Svoboda, eds.); Proc. Second Intern. Symp. Yeast Protoplasts (Brno, 1968), pp. 107–119. Brno: University J. E. Purkyně.
Thiéry, J.-P., 1967: Mise en évidence des polysaccharides sur coupes fines en microscopie électronique. J. Microscopie6, 987–1018.
Tokunaga, J., andS. Bartnicki-Garcia, 1971: Structure and differentiation of the cell wall ofPhytophthora, palmivora: cysts, hyphae and sporangia. Arch. Mikrobiol.79, 293–310.
Valk, P. van der, andJ. G. H. Wessels, 1973: Mitotic synchrony in multinucleateSchizophyllum protoplasts. Protoplasma78, 427–432.
Villanueva, J. R., andI. García-Acha, 1971: Production and use of fungal protoplasts. In: Methods in Microbiology (C. Booth, ed.), Vol. 4, pp. 665–718. New York: Academic Press.
Vries, O. M. H. de, andJ. G. H. Wessels, 1972: Release of protoplasts fromSchizophyllum commune by a lytic enzyme preparation fromTrichoderma viride, J. gen. Microbiol.73, 13–22.
— —, 1973: Release of protoplasts fromSchizopbyllum commune by combined action of purified α-1,3-glucanase and chitinase derived fromTrichoderma viride. J. gen. Microbiol.76, 319–330.
— —, 1975: Chemical analysis of cell wall regeneration and reversion of protoplasts fromSchizophyllum commune. Arch. Microbiol.102, 209–218.
Wessels, J. G. H., 1965: Morphogenesis and biochemical processes inSchizopbyllum commune Fr. Wentia13, 1–113.
—,D. R. Kreger, R. Marchant, B. A. Regensburg, andO. M. H. de Vries, 1972: Chemical and morphological characterization of the hyphal wall surface of tle basidiomyceteSchizophyllum commune. Biochim. biophys. Acta273, 346–358.
—,D. R. Kreger, R. Marchant, B. A. Regensburg, 1974: Enzymic degradation of septa in hyphal wall preparations from a monokaryon and a dikaryon ofSchizopbyllum commune. J. gen. Microbiol.83, 359–368.
- P.van derValk, and O. M. H. deVries, 1976: Wall synthesis by fungal protoplasts. In: Proceedings of the fourth International Symposium on Yeast and other Protoplasts (Nottingham, 1975). In press.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
van der Valk, P., Wessels, J.G.H. Ultrastructure and localization of wall polymers during regeneration and reversion of protoplasts ofSchizophyllum commune . Protoplasma 90, 65–87 (1976). https://doi.org/10.1007/BF01276480
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01276480