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Isolation of protoplasts by means of a “species-specific” autolysine inChlamydomonas

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Summary

Prior to fusion, gametes ofChlamydomonas reinhardii discard their cell walls. This naturally occuring phenomenon has provided the basis for a method of protoplast isolation from both gametes and vegetative cells within the genusChlamydomonas. When synchronized cultures of compatibleChlamydomonas gametes are mixed it is possible, after removal of the cells, to obtain a solution having a high cell wall lytic activity. That vegetative and gamete cells after treatment with this “gamete-autolysine” are indeed protoplasts has been proved by various light and electron microscopical methods.

The species specifity of this autolysine, its difference to the previously described “sporangialautolysine” (Schlösser 1966) and furthermore its use in the large scale production of protoplasts is also described. Since this wall autolysine is a factor produced by the cells themselves at a particular stage in their life cycle it represents a non-foreign agent in contrast to all other enzymic methods previously employed for protoplast isolation.

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References

  • Atkinson, A. W., B. E. S. Gunning, andP. C. L. John, 1972: Sporopollenin in the cell wall ofChlorella and other algae: Ultrastructure, chemistry and incorporation of14C-acetate, studied in synchronous cultures. Planta (Berl.)107, 1–32.

    Google Scholar 

  • Branton, D., 1971: Freeze-etching studies of membrane structure. Phil. Trans. Roy. Soc. (Lond.)B 261, 133–138.

    Google Scholar 

  • Bray, D. F., K. Nakamura, J. W. Costerton, andE. B. Wagenaar, 1974: Ultrastructure ofChlamydomonas eugametos as revealed by freeze-etching: Cell wall, plasmalemma and chloroplast membrane. J. Ultrastruct. Res.47, 125–141.

    Google Scholar 

  • Burczyk, J., H. Grzybek, J. Banás, andE. Banás, 1971: Presence of cellulase in the algaScenedesmus. Exp. Cell Res.63, 451–453.

    Google Scholar 

  • Claes, H., 1971: Autolyse der Zellwand bei den Gameten vonChlamydomonas reinhardii. Arch. Mikrobiol.78, 180–188.

    Google Scholar 

  • Cocking, E. C., 1972: Plant cell protoplasts—isolation and development. Ann. Rev. Plant Physiol.23, 29–50.

    Google Scholar 

  • — andP. K. Evans, 1973: The isolation of protoplasts. In: Plant tissue and cell culture, pp. 100–120. (H. E. Street, ed.). Oxford: Blackwell Sci. Publ.

    Google Scholar 

  • Davies, D. R., andV. Lyall, 1973: The assembly of a highly ordered component of the cell wall: The role of heritable factors and of physical structure. Molec. gen. Genetics124, 21–34.

    Google Scholar 

  • Deason, T. R., 1967:Chlamydomonas gymnogama, a new homothallic species with naked gametes. J. Phycol.3, 109–112.

    Google Scholar 

  • Franke, W. W., S. Krien, andR. M. Brown, Jr., 1969: Simultaneous glutaraldehydeosmium tetroxide fixation with postosmication, an improved fixation procedure for electron microscopy of plant and animal cells. Histochemie19, 162–164.

    Google Scholar 

  • Gabriel, M., 1970: Formation, growth, and regeneration of protoplasts of the green alga,Uronema gigas. Protoplasma70, 135–138.

    Google Scholar 

  • Hills, G. J., 1973: Cell wall assemblyin vitro fromChlamydomonas reinhardii. Planta (Berl.)115, 17–23.

    Google Scholar 

  • —,M. Gurney-Smith, andK. Roberts, 1973: Structure, composition, and morphogenesis of the cell wall ofChlamydomonas reinhardii. II. Electron microscopy and optical diffraction analysis. J. Ultrastruct. Res.43, 179–192.

    Google Scholar 

  • Horne, R. W., D. R. Davies, K. Norton, andM. Gurney-Smith, 1971: Electron microscope and optical diffraction studies on isolated cell walls fromChlamydomonas. Nature (Lond.)232, 493–495.

    Google Scholar 

  • Hoshaw, R. W., andH. Ettl, 1966:Chlamydomonas smithii sp. nov.—a chlamydomonad interfertile withChlamydomonas reinhardii. J. Phycol.2, 93–96.

    Google Scholar 

  • Klebs, G., 1896: Die Bedingungen der Fortpflanzung bei einigen Algen und Pilzen, pp. 424–438. Jena: G. Fischer.

    Google Scholar 

  • Kuhl, A., 1962: Zur Physiologie der Speicherung kondensierter anorganischer Phosphate. In: Beiträge zur Physiologie und Morphologie der Algen, pp. 157–164. Stuttgart: G. Fischer.

    Google Scholar 

  • — andH. Lorenzen, 1964: Handling and culturing ofChlorella. In: Methods in cell physiology. Vol. I, pp. 159–187. (D. M. Prescott, ed.). New York: Academic Press.

    Google Scholar 

  • Lorenzen, H., 1957: Synchrone Zellteilungen vonChlorella bei verschiedenen Licht-Dunkel-Wechseln. Flora (Jena)144, 473–496.

    Google Scholar 

  • Miller, D. H., D. T. A. Lamport, andH. Miller, 1972: Hydroxyproline heterooligosaccharides inChlamydomonas. Science176, 918–920.

    Google Scholar 

  • —,I. S. Mellman, D. T. A. Lamport, andM. Miller, 1974: The chemical composition of the cell wall ofChlamydomonas gymnogama and the concept of a plant cell wall protein. J. Cell Biol.63, 420–429.

    Google Scholar 

  • Pascher, A., 1943: Beiträge zur Morphologie der ungeschlechtlichen und geschlechtlichen Vermehrung der GattungChlamydomonas. Beih. Biol. Centralblatt Abt.A 62, 197–220.

    Google Scholar 

  • Potrykus, I., 1973: Isolation, fusion and culture of protoplasts ofPetunia. In: Yeast, mould and plant protoplasts, pp. 319–332. (J. R. Villanueva, I. García-Acha, S. Gáscon, andF. Uruburu, eds.). London-New York: Academic Press.

    Google Scholar 

  • Roberts, K., 1974: Crystalline glycoprotein cell walls of algae: their structure, composition and assembly. Phil. Trans. Roy. Soc. (Lond.)B 268, 129–146.

    Google Scholar 

  • —,M. Gurney-Smith, andG. J. Hills, 1972: Structure, composition and morphogenesis of the cell wall ofChlamydomonas reinhardii. I. Ultrastructure and preliminary chemical analysis. J. Ultrastruct. Res.40, 599–613.

    Google Scholar 

  • Ruesink, A. W., 1971: Protoplasts of plant cells. In: Methods in enzymology, Vol. 23, pp. 197–209. (A. San Pietro, ed.). London-New York: Academic Press.

    Google Scholar 

  • Sager, R., andS. Granick, 1953: Nutritional studies withChlamydomonas reinhardii. Ann. N.Y. Acad. Sci.56, 831.

    Google Scholar 

  • — — 1954: Nutritional control of sexuality inChlamydomonas reinhardii. J. gen. Physiol.37, 729–742.

    Google Scholar 

  • Schlösser, U., 1966: Enzymatisch gesteuerte Freisetzung von Zoosporen beiChlamydomonas reinhardii Dangeard in Synchronkultur. Arch. Mikrobiol.54, 129–159.

    Google Scholar 

  • Skuja, H., 1963: Die geschlechtliche Reproduktion einer Monade aus dem Formenkreis vonChlamydomonas tecta. Svensk. Bot. Tidskr.57, 243–252.

    Google Scholar 

  • Van der Woude, W. J., D. J. Morré, andC. E. Bracker, 1971: Isolation and characterization of secretory vesicles in germinating pollen ofLilium longiflorum. J. Cell Sci.8, 331–351.

    Google Scholar 

  • Villanueva, J. R., andI. García-Acha, 1971: Production and use of fungal protoplasts. In: Methods in microbiology. Vol. 4, pp. 665–718. (C. Booth, ed.). London-New York: Academic Press.

    Google Scholar 

  • Villanueva, J. R., I. García-Acha, S. Gáscon, andF. Uruburu (eds.), 1973: Yeast, mould and plant protoplasts. London-New York: Academic Press.

    Google Scholar 

  • Wiese, L., 1965: On sexual agglutination and mating-type substances (gamones) in isogamous heterothallicChlamydomonads. I. Evidence of the identity of the gamones with surface components responsable for sexual flagellar contact. J. Phycol.1, 46–54.

    Google Scholar 

  • — 1974: Nature of sex specific glycoprotein agglutinins inChlamydomonas. Ann. N.Y. Acad. Sci.234, 383–395.

    Google Scholar 

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Schlösser, U.G., Sachs, H. & Robinson, D.G. Isolation of protoplasts by means of a “species-specific” autolysine inChlamydomonas . Protoplasma 88, 51–64 (1976). https://doi.org/10.1007/BF01280359

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  • DOI: https://doi.org/10.1007/BF01280359

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