Archives of Microbiology

, Volume 108, Issue 2, pp 189–194 | Cite as

Synchronized cultures of a cell wall-less mutant of Chlamydomonas reinhardii

  • Torleiv Lien
  • Gjert Knutsen
Article

Abstract

Synchronization and synchronous growth of a cell wall-less mutant of Chlamydomonas reinhardii have been described. The following growth conditions were used: A modified Sueokas' “high salt minimal medium”, 14∶10 h light-dark cycle, growth temperature 30°C, light intensity 12–18 Klux and dilution of the culture at the end of the dark to a constant cell density of 1.0·106 cells/ml. The time course of increase and distribution of cell volume, cytoplasmic and nuclear division, release of motile cells after the division period and accumulation of DNA, RNA and protein are reported. These mutant cells did not make any sporangium in which the dividing cells were kept as a unit inside a mother cell wall. However, they usually adhered during the period of division, thus making clumps containing 2, 4 and 8 cells. Several of these cell clumps dissolved releasing either single or couples of 2 and 4 cells. After the end of division the cells became flagellated and motile and thereby releasing themselves from the aggregate.

Key words

Chlamydomonas reinhardii Synchronous growth Cell wall-less mutant 

Non-Standard Abbreviations

AWV

average weighed cell volume

MM

minimal medium

HSM

high salt medium

TCA

trichloroacetic acid

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Baker, A. L., Schmidt, R. R.: Further studies on the intracellular distribution of phosphorus during synchronous growth of Chlorella pyrenoidosa. Biochim. biophys. Acta (Amst.) 82, 336–342 (1964)Google Scholar
  2. Bolognani, L., Coppi, G., Zambotti, V.: Determination of pentoses. Experientia (Basel) 17, 67–68 (1961)Google Scholar
  3. Burton, K.: A study of the conditions and mechanism of diphenyl-amine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem. J. 62, 315–323 (1956)Google Scholar
  4. Cattolico, R. A., Senner, J. W., Jones, R. F.: Changes in cytoplasmic and chloroplast ribosomal ribonucleic acid during the cell cycle of Chlamydomonas reinhardtii. Arch. Biochem. Biophys. 156, 58–65 (1973)Google Scholar
  5. Chiang, K., Sueoka, N.: Replication of chloroplast DNA in Chlamydomonas reinhardi during vegetative cell cycle: its mode and regulation. Proc. nat. Acad. Sci. (Wash.) 57, 1506–1513 (1967)Google Scholar
  6. Davies, D. R., Plaskitt, A.: Genetical and structural analyses of cell-wall formation in Chlamydomonas reinhardi. Genet. Res. 17, 33–43 (1971)Google Scholar
  7. Gould, R. R.: The basal bodies of Chlamydomonas reinhardtii. J. Cell Biol. 65, 65–74 (1975)Google Scholar
  8. Holm-Hansen, O., Sutdiffe, W. H., Jonathan, S.: Measurement of deoxyribonucleic acid in the ocean and its ecological significance. Limnol. Oceanogr. 13, 507–514 (1968)Google Scholar
  9. Hyams, J., Davies, D. R.: The induction and characterisation of cell wall mutants of Chlamydomonas reinhardi. Mutation Res. 14, 381–389 (1972)Google Scholar
  10. Kissane, J. M., Robbins, E.: The fluorometric measurement of deoxyribonucleic acid in animal tissues with special reference to the central nervous system. J. biol. Chem. 233, 184–188 (1958)Google Scholar
  11. Knutsen, G.: Repressed and derepressed synthesis of phosphatases during synchronous growth of Chlorella pyrenoidosa. Biochim. biophys. Acta (Amst.) 161, 205–214 (1968)Google Scholar
  12. Kuhl, A., Lorenzen, H.: Handling and culturing of Chlorella. In: Methods in cell physiology, Vol. 1 (D. M. Prescott, ed.), pp. 159–188. New York-London: Academic Press 1964Google Scholar
  13. Lien, T., Knutsen, G.: Phosphate as a control factor in cell division of Chlamydomonas reinhardti, studied in synchronous culture. Exp. Cell Res. 78, 79–88 (1973)Google Scholar
  14. Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurements with the Folin Phenol Reagent. J. biol. Chem. 193, 265–275 (1951)Google Scholar
  15. Roberts, K., Gurney-Smith, M., Hills, J. G.: Structure, composition and morphogenesis of the cell wall of Chlamydomonas reinhardti. I. Ultrastructure and preliminary chemical analysis. J. Ultrastruct. Res. 40, 599–613 (1972)Google Scholar
  16. Robreau, G., Le Gal, Y.: Conditions for isolating chromatin from Chlamydomonas reinhardtii CW15. Hébd. Séances Acad. Sci., Ser. D 279, 1923–1926 (1974)Google Scholar
  17. Senger, H.: Untersuchung zur Synchronisierung von Chlorella-Kulturen. Arch. Mikrobiol. 40, 47–72 (1961)Google Scholar
  18. Sueoka, N., Chiang, K. S., Kates, R. J.: Deoxyribonucleic acid replication in meiosis of Chlamydomonas reinhardi. J. molec. Biol. 25, 47–66 (1967)Google Scholar
  19. Wanka, F.: Die Bestimmung der Nucleinsäuren in Chlorella-Kulturen. Planta (Berl.) 58, 594–609 (1962)Google Scholar

Copyright information

© Springer-Verlag 1976

Authors and Affiliations

  • Torleiv Lien
    • 1
    • 2
  • Gjert Knutsen
    • 1
    • 2
  1. 1.Institute for General MicrobiologyUniversity of BergenBergen-UniversitetetNorway
  2. 2.Botanical LaboratoryUniversity of BergenBergen-UniversitetetNorway

Personalised recommendations