Summary
By culturing of callus tissue originating from root explants of Petroselinum sativum in a synthetic liquid medium under aeration, freely suspended single cells and small clusters consisting of mostly five cells were obtained. The rapidly dividing cells did not exhibit any morphogenesis. Their nucleic acid metabolism was investigated by pulse experiments with 32P-orthophosphate. Rapidly labelled RNA was prominently found associated with high molecular RNA. During the fractionation of the total nucleic acids on MAK columns it was eluted after the ribosomal RNA components. Its base ratio, however, differed from the latter in that the AMP content was higher than the GMP content. Sucrose gradient centrifugation and polyacrylamide gel electrophoresis resulted in the separation of the ribosomal RNA from the rapidly labelled RNA, thus proving the higher molecular weight of the latter. Based upon the migration in the gel a sedimentation coefficient of approximately 32S was calculated. The possible function of the heavy rapidly labelled RNA component as precursor of ribosomal RNA is discussed.
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Literatur
Bergmann, L.: Wachstum grüner Suspensionskulturen von Nicotiana tabacum var. “Samsun” mit CO2 als Kohlenstoffquelle. Planta (Berl.) 74, 243–249 (1967).
— Berger, Ch.: Farblicht und Plastidendifferenzierung in Zellkulturen von Nicotiana tabacum var. “Samsun”. Planta (Berl.) 69, 58–69 (1966).
Bishop, D. H. L., Claybrook, J. R., Spiegelman, S.: Electrophoretic separation of viral nucleic acids on polyacrylamide gels. J. molec. Biol. 26, 373–387 (1967).
Brawerman, G., Hufnagel, D. A., Chargaff, E.: On the nucleic acids of green and colorless Euglena gracilis. Isolation and composition of DNA and tRNA. Biochim. biophys. Acta (Amst.) 61, 340–345 (1962).
Cherry, J. H., Van Huystee, R.: Effects of 5-fluorouracil on photoperiodic induction on nucleic acid metabolism of Xanthium. Plant Physiol. 40, 987–993 (1965).
Ewing, E. E., Cherry, J. H.: Presence of AMP-rich RNA in countercurrent distribution and column chromatography fractions of pea root nucleic acids. Biochim. biophys. Acta (Amst.) 161, 331–340 (1968).
Hayashi, M. N., Hayshi, M., Spiegelman, S.: Chromatographic separation of annealed and enzymatically synthesized RNA-DNA hybrids. Biophys. J. 5, 231–246 (1965).
Hemleben-Vielhaben, V.: Characterization of rapidly labelled nucleic acids in tissues of plant seedlings. Z. Naturforsch. 21b, 983–992 (1966).
Hock, B.: Nature of a rapidly labelled RNA fraction described in higher plant system. Plant Physiol. 42, 1149–1152 (1967).
Ingle, J., Key, J. L.: A comparative evaluation of the synthesis of DNA-like RNA in excised and intact plant tissues. Plant Physiol. 40, 1212–1219 (1965).
Koblitz, H., Hagen, I.: Vergleichende Untersuchungen über das Wachstum isolierter Karottengewebe auf halbsynthetischen Substraten und auf einem neuen vollsynthetischen Medium. Flora (Jena) 152, 447–457 (1962).
Kirby, K. S.: Isolation and characterization of ribosomal ribonucleic acid. Biochem. J. 96, 266–269 (1965).
Kuhl, A., Lorenzen, H.: Handling and culturing of Chlorella. In: Methods in cell physiology (ed. D. M. Prescott), vol. I., p. 159. New York and London: Academic Press 1964.
Lin, C. Y., Key, J. L., Bracker, C. E.: Association of D-RNA with polyribosomes in soy bean root. Plant Physiol. 41, 976–982 (1966).
Loening, U. E.: The fracionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis. Biochem. J. 102, 251–257 (1967).
—: RNA structure and metabolism. Ann. Rev. Plant. Physiol. 19, 37–70 (1968).
— In: Chromatographic and electrophoretic techniques (ed. I. Smith), vol. II, p. 437. Kingswood, Engl.: W. Heinemann (Medical Books) Ltd. 1968b.
— Ingle, J.: Diversity of RNA components in green plant tissues. Nature (Lond.) 215, 363–367 (1967).
Lonberg-Holm, K. K.: Nucleic acid synthesis in seedlings. Nature (Lond.) 213, 454–457 (1967).
Mandell, I. D., Hershey, A. D.: A fractionating column for analysis of nucleic acids. Analyt. Chem. 1, 66–74 (1960).
Miller, A. O. A., Dhont, E., De Vreese, A. M., Van Nimmen, L.: Study of messenger like RNA extracted from HeLa cells polysomes. I. Sedimentation and chromatographic data. Biochem. biophys. Res. Commun. 33, 702–708 (1968).
Murashige, T., Skoog, F.: A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 15, 473–497 (1962).
Reinhard, E.: Probleme der Produktion von Arzneistoffen durch pflanzliche Gewebekulturen. Dtsch. Apoth.-Ztg 107, 1201–1207 (1967).
Richter, G., Senger, H.: Extraktion und chromatographische Auftrennung der Nukleinsäuren aus Photosynthese-Organismen. Biochim. biophys. Acta (Amst.) 87, 502–505 (1964).
Ruppel, H. G.: Nucleinsäuren in Chloroplasten. II. Isolierung und Charakterisierung der Chloroplasten-Ribosomen von Antirrhinum majus. Z. Naturforsch. 23b, 997–1004 (1968).
Scherrer, K., Latham, H., Darnell, J. E.: Demonstration of an unstable RNA and of a precursor to ribosomal RNA in HeLa cells. Proc. nat. Acad. Sci. (Wash.) 49, 240–248 (1963).
Taber, R. L., Vincent, W. S.: The synthesis and processing of ribosomal RNA precursor molecules in yeast. Biochim. biophys. Acta (Amst.) 186, 317–325 (1969).
Weinberg, R. A., Loening, U., Willems, M., Penman, S.: Acrylamide gel electrophoresis of HeLa cells nucleolar RNA. Proc. nat. Acad. Sci. (Wash.) 58, 1088–1095 (1967).
Yoshikawa-Fukada, M., Fukada, T., Kawade, Y.: Characterization of rapidly labelled ribonucleic acid of animal cells in culture. Biochim. biophys. Acta (Amst.) 103, 383–398 (1965).
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Seitz, U., Richter, G. Isolierung und Charakterisierung schnell markierter, hochmolekularer RNS aus frei suspendierten Calluszellen der Petersilie (Petroselinum sativum). Planta 92, 309–326 (1970). https://doi.org/10.1007/BF00385098
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DOI: https://doi.org/10.1007/BF00385098