Abstract
In cultures of isolated mesophyll cells ofZinnia elegans, transdifferentiation into tracheary elements is induced by a combination of auxin and cytokinin and is blocked by inhibitors of DNA synthesis and poly (ADP-ribose) synthesis. During transdifferentiation, a very low level of synthesis of nuclear DNA was found in some cultured cells by microautoradiography after pulse-labeling with [3H]thymidine. Density profiles of nuclear DNA that had been double-labeledin vivo with bromodeoxyuridine (BrdU) and [3H]thymidine indicated that this DNA synthesis was repair-type synthesis. The sedimentation velocity of nucleoids increased during the culture of isolated mesophyll cells and the increase was dependent on phytohormones. This phenomenon may reflect the rejoining of DNA strand breaks after repair-type DNA synthesis during transdifferentiation. Treatment of cells with inhibitors of DNA synthesis or of poly(ADP-ribose) synthesis prevented the increase in the sedimentation velocity of nucleoids. The data suggest the involvement of DNA-repair events in the transdifferentiation of mesophyll cells into tracheary elements.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- APC:
-
aphidicolin
- BA:
-
benzyladenine
- BrdU:
-
5-bromodeoxyuridine
- EDTA:
-
ethylenediaminetetraacetic acid
- EtBr:
-
ethidium bromide
- Mes:
-
2-(N-morpholino)-ethanesulfonic acid
- NAA:
-
1-naphthaleneacetic acid
References
Althaus, F.R., Lawrence, S.D., He, Y.-Z., Sattler, G.L., Tsukada, Y. andPitot, H.C. 1982. Effects of altered [ADP-ribose]n metabolism on expression of fetal functions by adult hepatocytes. Nature300: 366–368.
Althaus, F.R., Lawrence, S.D., Sattler, G.L. andPitot, H.C. 1982. ADP-ribosyltransferase activity in cultured hepatocytes. J. Biol. Chem.257: 5528–5535.
Anet, b R. andStrayer, D.R. 1969. Density gradient relaxation: a method for preparative buoyant density separations of DNA. Biochem. Biophys. Res. Commun.34: 328–334.
Clark, J.B., Ferris, G.M. andPinder, S. 1971. Inhibition of nuclear NAD nucleosidase and poly ADP-ribose polymerase activity from rat liver by nicotinamide and 5'-methyl nicotinamide. Biochim. Biophys. Acta238: 82–85.
Cleaver, J.E. 1983. Structure of repaired sites in human DNA synthesized in the presence of inhibitors of DNA polymerases alpha and beta in human fibroblasts. Biochim. Biophys. Acta739: 301–311.
Collins, A. 1983. DNA repair in ultraviolet-irradiated HeLa cells is disrupted by aphidicolin: the inhibition of repair need not imply the absence of repair synthesis. Biochim. Biophys. Acta741: 341–347.
Cook, P.R. andBrazell, I.A. 1976. Conformational constraints in nuclear DNA. J. Cell Sci.22: 287–302.
Farzaneh, F., Shall, S. andJohnstone, A.P. 1985. The dynamic nature of DNA-strand breaks present in differentiating muscle cells and quiescent lymphocytes. FEBS Lett.189: 62–66.
Farzaneh, F., Zalin, R., Brill, D. andShall, S. 1982. DNA strand breaks and ADP-ribosyltransferase activation during cell differentiation. Nature300: 362–366.
Fukuda, H. andKomamine, A. 1980. Establishment of an experimental system for the study of tracheary element differentiation from single cells isolated from the mesophyll ofZinnia elegans. Plant Physiol.65: 57–60.
Fukuda, H. andKomamine, A. 1981a. Relationship between tracheary element differentiation and the cell cycle in single cells isolated from the mesophyll ofZinnia elegans. Physiol. Plant.52: 423–430.
Fukuda, H. andKomamine, A. 1981b. Relationship between tracheary element differentiation and DNA synthesis in single cells isolated from the mesophyll ofZinnia elegans: analysis by inhibitors of DNA synthesis. Plant Cell Physiol.22: 41–49.
Hartley, J.A., Gibson, N.W., Zwelling, L.A. andYuspa, S.H. 1985. Association of DNA strand breaks with accelerated terminal differentiation in mouse epidermal cells exposed to tumor promoters. Cancer Res.45: 4864–4870.
Hawkins, S.W. andPhillips, R. 1983. 3-Aminobenzamide inhibits tracheary element differentiation but not cell division in cultured explants ofHelianthus tuberosus. Plant Sci. Lett.32: 221–224.
Huberman, J.A. 1981. New views of the biochemistry of eukaryotic DNA replication revealed by aphidicolin, an unusual inhibitor of DNA polymerase α. Cell23: 647–648.
Ikegami, S., Taguchi, T., Ohashi, M., Oguro, M., Nagano, H. andMano, Y. 1978. Aphidicolin prevents mitotic cell division by interfering with the activity of DNA polymerase-α. Nature275: 458–460.
Johnstone, A.P. andWilliams, G.T. 1982. Role of DNA breaks and ADP-ribosyltransferase activity in eukaryotic differentiation demonstrated in human lymphocytes. Nature300: 368–370.
Mattern, M.R., Paone, R.F. andDay III, R.S. 1982. Eukaryotic DNA repair is blocked at different steps by inhibitors of DNA topoisomerases and of DNA polymerases α and β. Biochim. Biophys. Acta697: 6–13.
Morita, T., Tsutsui, Y., Nishiyama, Y., Nakamura, H. andYoshida, S. 1982. Effects of DNA polymerase inhibitors on replicative and repair DNA synthesis in ultraviolet-irradiated HeLa cells. Int. J. Radiat. Biol.42: 471–480.
Phillips, R. andHawkins, S.W. 1985. Characteristics of the inhibition of induced tracheary element differentiation by 3-aminobenzamide and related compounds. J. Exp. Bot.36: 119–128.
Rogers, S.O. andBendich, A.J. 1985. Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues. Plant Mol. Biol.5: 69–76.
Sala, F., Magnien, E., Galli, M.G., Daschaert, X., Pedrali-Noy, G. andSpadari, S. 1982. DNA repair synthesis in plant protoplasts is aphidicolin-resistant. FEBS Lett.138: 213–217.
Smith, C.A. andOkumoto, D.S. 1984. Nature of DNA repair synthesis resistant to inhibitors of polymerase α in human cells. Biochemistry23: 1383–1391.
Smith, C.A., Cooper, P.K. andHanawalt, P.C. 1981. Measurement of repair replication by equilibrium sedimentation.In E.C. Friedberg and P.C. Hanawalt, eds., DNA Repair, a Laboratory Manual of Research Procedures, Vol. 1 Part B. Marcel Dekker, New York, pp. 289–305.
Stoilov, L.M., Zlatanova, J.S., Vassileva, A.P., Ivanchenko, M.G., Krachmarov, Ch.P. andGenchev, D.D. 1988. Supercoils in plant DNA: nucleoid sedimentation studies. J. Cell Sci.89: 243–252.
Sugiyama, M., Fukuda, H. andKomamine, A. 1986. Effects of nutrient limitation and gamma-irradiation on tracheary element differentiation and cell division in single mesophyll cells ofZinnia elegans. Plant Cell Physiol.27: 601–606.
Sugiyama, M. andKomamine, A. 1987. Effect of inhibitors of ADP-ribosyltransferase on the differentiation of tracheary elements from isolated mesophyll cells ofZinnia elegans. Plant Cell Physiol.28: 541–544.
Sugiyama, M., Fukuda, H. andKomamine, A. 1990. Characteristics of the inhibitory effect of 5-fluorodeoxyuridine on cytodifferentiation into tracheary elements of isolated mesophyll cells ofZinnia elegans. Plant Cell Physiol.31: 61–67.
Suglyama, M., Fukuda, H. andKomamine, A. 1994. Characterization of the inhibitory effect of aphidicolin on transdifferentiation into tracheary elements of isolated mesophyll cells ofZinnia elegans. Plant Cell Physiol.35: 519–522.
Th'ng, J.P.H. andWalker, I.G. 1985. Excision repair of DNA in the presence of aphidicolin. Mutation Res.165: 139–150.
Wang, T.S.-F. 1991. Eukaryotic DNA polymerases. Annu. Rev. Biochem.60: 513–552.
Willmitzer, L. andWagner, K.G. 1981. The isolation of nuclei from tissue-cultured plant cells. Exp. Cell Res.135: 69–77.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sugiyama, M., Yeung, E.C., Shoji, Y. et al. Possible involvement of DNA-repair events in the transdifferentiation of mesophyll cells ofZinnia elegans into tracheary elements. J. Plant Res. 108, 351–361 (1995). https://doi.org/10.1007/BF02344360
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02344360