Skip to main content
SpringerLink
Log in

DNA profile of the spore of blastocladiella emersonii: Evidence for γ-particle DNA

  • Published:
Archiv für Mikrobiologie Aims and scope Submit manuscript

Summary

Procedures were developed for isolating DNA from zoospores of Blastocladiella emersonii, critical steps being the use of non-frozen cells, enzymatic elimination of polysaccharide, and a combined treatment with T1RNAse and pancreatic RNAse for complete removal of RNA. Methods for isolating γ-particles from spores were also established. Extraction of DNA directly from purified γ-particles substantiated previous assumptions that they contain DNA. Heat denaturation and preparative CsCl equilibrium sedimentation studies of whole spore DNA revealed the presence of four distinct components, I–IV, with buoyant densities of 1.731, 1.715, 1.705, and 1.687 g/cm3. The major species appeared to be nuclear chromatin. Nucleolar and mitochondrial sites were tentatively assigned to species II and III, respectively. Evidence was presented that species IV resided in γ-particles. Some possible roles for γ-particle DNA were discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

EGTA:

ethyleneglycol-bis (B-aminoethyl ether)-N,N′-tetracetic acid

EDTA:

ethylenediaminetetracetic acid

TCA:

trichloroacetic acid

SDS:

sodium dodecyl sulfate

SSC:

0.15 M NaCl and 15 mM Na citrate

References

  • Burton, K.: A study of the condition and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem. J. 62, 315–323 (1956).

    Google Scholar 

  • Cantino, E. C.: Morphogenesis in aquatic fungi, pp. 283–337. In: G. C. Ainsworth and A. S. Sussman (Eds.): The fungi, Vol. 2. New York: Academic Press 1966.

    Google Scholar 

  • —, Horenstein, E. A.: Cytoplasmic exchange without gametic copulation in the water mold Blastocladiella emersonii. Amer. Nat. 88, 143–154 (1954).

    Google Scholar 

  • ——: Gamma and the cytoplasmic control of differentiation in Blastocladiella. Mycologia (N. Y.) 48, 443–446 (1956).

    Google Scholar 

  • —, Lovett, J. S., Leak, L. V., Lythgoe, J.: The single mitochondrion, fine structure, and germination of the spore of Blastocladiella emersonii. J. gen. Microbiol. 31, 393–404 (1963).

    Google Scholar 

  • —, Mack, J. P.: Form and function in the zoospore of Blastocladiella emersonii. I. The γ-particle and satellite ribosome package. Nova Hedwigia 18, 115–147 (1969).

    Google Scholar 

  • —, Truesdell, L. C., Shaw, D. S.: Life history of the motile spore of Blastocladiella emersonii: A study in cell differentiation. J. Elisha Mitchell Sci. Soc. 84, 125–146 (1968).

    Google Scholar 

  • Comb, D. G., Brown, R., Katz, S.: The nuclear DNA and RNA components of the aquatic fungus Blastocladiella emersonii. J. molec. Biol. 8, 781–789 (1964).

    Google Scholar 

  • Coudray, Y., Quetier, F., Guille, E.: New compilation of satellite DNAs. Biochim. biophys. Acta 217, 259–267 (1970).

    Google Scholar 

  • Flamm, W. G., Birnstiel, M. L., Walker, P. M. B.: Preparation and fractionation, and isolation of single strands, of DNA by isopycnic ultracentrifugation in fixed-angle rotors, pp. 125–155. In: G. D. Birnie and S. M. Fox (eds.) Subcellular components. London: Butterworths 1969.

    Google Scholar 

  • Hruban, Z., Rechcigl, M.: Microbodies and related particles. New York: Academic Press 1969.

    Google Scholar 

  • Knittel, M. D., Black, C. H., Sandine, W. E., Fraser, Dorothy K.: Use of normal probability paper in determining thermal melting values of deoxyribonucleic acid. Canad. J. Microbiol. 14, 239–245 (1968).

    Google Scholar 

  • Lee, L. F., Boezi, J. A.: Characterization of bacteriophage gh-1 for Pseudomonas putida. J. Bact. 92, 1821–1827 (1966).

    Google Scholar 

  • Lessie, P. E., Lovett, J. S.: Ultrastructural changes during sporangium formation and zoospore differentiation in Blastocladiella emersonii. Amer. J. Bot. 55, 220–236 (1968).

    Google Scholar 

  • Lovett, J. S.: Chemical and physical characterization of “nuclear caps” isolated from Blastocladiella zoospores. J. Bact. 85, 1235–1246 (1963).

    Google Scholar 

  • —: Aquatic fungi, pp. 341–358. In: F. H. Wilt and N. K. Wessells (eds.): Methods in developmental biology. New York: T. Y. Crowell Co. 1967.

    Google Scholar 

  • —: Reactivation of ribonucleic acid and protein synthesis during germination of Blastocladiella zoospores and the role of the ribosomal nuclear cap. J. Bact. 96, 962–969 (1968).

    Google Scholar 

  • Maden, B. E. H.: Ribosome formation in animals cells. Nature (Lond.) 219, 685–689 (1968).

    Google Scholar 

  • Mandel, M., Igambi, L., Bergendahl, J., Dodson, M. L. Jr., Scheltgen, E.: Correlation of melting temperature and cesium chloride buoyant density of bacterial deoxyribonucleic acid. J. Bact. 101, 333–338 (1970).

    Google Scholar 

  • Marmur, J.: A procedure for the isolation of deoxyribonucleic acid from microorganisms. J. molec. Biol. 3, 208–218 (1961).

    Google Scholar 

  • —, Doty, P.: Determination of the base composition of deoxyribonucleic acid from its thermal denaturation temperature. J. molec. Biol. 5, 109–118 (1962).

    Google Scholar 

  • Matsumae, A., Myers, R. B., Cantino, E. C.: Comparative numbers of γ-particles in the flagellate cells of various species and mutants of Blastocladiella. J. gen. appl. Microbiol. 16, 443–453 (1970).

    Google Scholar 

  • Miller, O. L., Jr., Beatty, B. R.: Extrachromosomal nucleolar genes in amphibian oocytes. Genetics Suppl. 61, 1, 133–143 (1969).

    Google Scholar 

  • Nass, M. M. K.: Mitochondrial DNA: Advances, problems, and goals. Science 165, 25–35 (1969a).

    Google Scholar 

  • ——: Mitochondrial DNA. II. Structure and physicochemical properties of isolated DNA. J. molec. Biol. 42, 529–545 (1969b).

    Google Scholar 

  • Schiff, J. A., Epstein, H. T.: The continuity of the chloroplast in Euglena, pp. 131–189. In: M. Locke (ed.): The Twenty-Fourth Sympos. Soc. Dev. Biology. New York: Academic Press 1965.

    Google Scholar 

  • Schildkraut, C. L., Marmur, J., Doty, P.: Determination of the base composition of deoxyribonucleic acid from its buoyant density in CsCl. J. molec. Biol. 4, 430–443 (1962).

    Google Scholar 

  • Schmid, P., Schmid, C., Brodie, D. C.: The determination of the total deoxyribose of deoxyribonucleic acid. J. biol. Chem. 238, 1068–1072 (1963).

    Google Scholar 

  • Schneider, W. C.: Determination of nucleic acids in tissues by pentose analysis, pp. 680–682. In: S. P. Colowick and N. O. Kaplan (eds.): Methods in enzymology, vol. 3. New York: Academic Press 1957.

    Google Scholar 

  • Shaw, D. S., Cantino, E. C.: An albino mutant of Blastocladiella emersonii: Comparative studies of zoospore behaviour and fine structure. J. gen. Microbiol. 59, 369–382 (1969).

    Google Scholar 

  • Sober, H. A. (ed.): Handbook of biochemistry. Cleveland, Ohio: The Chemical Rubber Co. 1968.

    Google Scholar 

  • Soll, D. R., Sonneborn, D. R.: Zoospore germination in Blastocladiella emersonii: Cell differentiation without protein synthesis? Proc. nat. Acad. Sci. (Wash.) 68, 459–463 (1971).

    Google Scholar 

  • Srb, A. M.: Extrachromosomal heredity in fungi, pp. 191–211, In: M. Locke (ed.): The Twenty-Fourth Sympos. Soc. for Dev. Biol. New York: Academic Press 1965.

    Google Scholar 

  • Storck, R., Alexopoulos, C. J.: Deoxyribonucleic acid of fungi. Bact. Rev. 34, 126–154 (1970).

    Google Scholar 

  • Truesdell, L. C., Cantino, E. C.: Decay of γ-particles in germinating zoospores of Blastocladiella emersonii. Arch. Mikrobiol. 70, 378–392 (1970).

    Google Scholar 

  • ——: The induction and early events of germination in the zoospore of Blastocladiella emersonii. In: A. Monroy and A. A. Moscona (eds.): Current topics in dev. Biol., vol. 6. New York: Academic Press 1971 (in press).

    Google Scholar 

  • Van Bruggen, E. F. J., Runner, C. M., Borst, P., Ruttenberg, G. J. C. M., Kroon, A. M., Schuurmans Stekhoven, F. M. A. H.: Mitochondrial DNA: III. Electron microscopy of DNA released from mitochondria by osmotic shock. Biochim. biophys. Acta (Amst.) 161, 402–414 (1968).

    Google Scholar 

  • Villa, V. D., Storck, R.: Nucleotide composition of nuclear and mitochondrial deoxyribonucleic acid of fungi. J. Bact. 96, 184–190 (1968).

    Google Scholar 

  • Vinograd, J., Hearst, J. E.: Equilibrium sedimentation of macromolecules and viruses in a density gradient, p. 372, In: L. Zechmeister (ed.): Fortschr. Chem. org. Naturstoffe, Bd. 20. Vienna: Springer 1962.

    Google Scholar 

  • Wyatt, G. R.: Recognition and estimation of 5-methyl cytosine in nucleic acids. Biochem. J. 48, 581–584 (1951).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Botany and Plant Pathology, Michigan State University, 48823, East Lansing, Michigan, USA

    R. B. Myers & E. C. Cantino

Authors
  1. R. B. Myers
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. C. Cantino
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Myers, R.B., Cantino, E.C. DNA profile of the spore of blastocladiella emersonii: Evidence for γ-particle DNA. Archiv. Mikrobiol. 78, 252–267 (1971). https://doi.org/10.1007/BF00424898

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00424898

Keywords

Search

Navigation