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Hormone-induced caulogenesis in long-term tobacco cell lines and its effect on nuclear DNA content

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Summary

Manipulation of exogenous auxin and cytokinin levels during sequential subculture in vitro has been used to induce caulogenesis in several long-term tobacco cell lines. Concurrently, tissue samples at various stages of caulogenesis have been examined for nuclear DNA content. While a variety of hormone regimes permitted caulogenesis, extremely high (122.95 μM) cytokinin levels and extremely low (0.285 μM) auxin levels generally gave optimal response. For three lines, caulogenesis was accompanied by a progressive decrease in nuclear DNA content beyond that due to the loss of polyploid cells. In one line, however, DNA content remained stable during regeneration, perhaps reflecting the acquisition of a stably adapted aneuploid karyotype. Both caulogenic response and amount of nuclear DNA were affected by changes in the culture medium. The progressive nature of the observed changes in DNA content is inconsistent with a single-step selection for euploid competent cells. Alternative models postulating either progressive selection for euploidy, or the regulation of karyotype are proposed to explain the results.

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Literature

  • Berlyn, G.P.; Miksche, J.P. (1976): Botanical Microtechnique and Cytochemistry. Ames: The Iowa State Univers. Press

    Google Scholar 

  • Brossard, D. (1975): La néoformation de bourgeons vegetatifs a partir de la moelle du tabac (Nicotiana tabacum L. cv. ‘Wisconsin 38’) cultivée in vitro. Analyse cytochemique, historadiographique, et cytophotométrique. Ann. Sci. Nat. Bot. Biol. Veg. 16, 43–150

    Google Scholar 

  • Dhillon, S.S.; Berlyn, G.P.; Miksche, J.P. (1977): Requirement of an internal standard for microspectrophotometric measurements of DNA. Amer. J. Bot. 64, 117–121

    Google Scholar 

  • Doerschug, E.B.; Miksche, J.P.; Palmer, R.G. (1978): DNA content, ribosomal-RNA gene number, and protein content in soybeans. Can. J. Genet. Cytol. 20, 531–538

    Google Scholar 

  • Filner, P. (1965): Semi-conservative replication of DNA in a higher plant cell. Expt. Cell Res. 39, 33–39

    Google Scholar 

  • Flashman, S.M.; Filner, P. (1978): Selection of tobacco cell lines resistant to selenoamino acids. Plant Sci. Let. 13, 219–229

    Google Scholar 

  • Heimer, Y.M.; Filner, P. (1970): Regulation of the nitrate assimilation pathway of cultured tobacco cells, II. Properties of a variant cell line. Biochim. Biophys. Acta. 215, 152–165

    Google Scholar 

  • Melchers, G.; Sacristan, M.D. (1977): Somatic hybridization of plants by fusion of protoplasts II. The chromosome numbers of somatic hybrid plants of 4 different fusion experiments. In: La Culture de Tissus et des Cellules de Végétaux, pp. 169–177. Paris: Masson

    Google Scholar 

  • Melchers, G. (1977): Microbial techniques in somatic hybridization by fusion of protoplasts. In: Intern. Cell Biology 1976–1977. (eds.: Brinkley, B.; Parker, K.R.), pp. 207–213. New York: The Rockefeller Univers. Press

    Google Scholar 

  • Murashige, T.; Nakano, R. (1965): Morphogenic behavior of tobacco tissue cultures and implication of plant senescence. Amer. Jour. Bot. 52, 819–827

    Google Scholar 

  • Murashige, T.; Nakano, R. (1967): Chromosome complement as a determinant of the morphogenic potential of tobacco cells. Amer. Jour. Bot 54, 963–967

    Google Scholar 

  • Murashige, T.; Skoog, F. (1962): A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol. Plant. 15, 473–497

    Google Scholar 

  • Ogura, H. (1976): The cytological chimeras in original regenerates from tobacco tissue cultures and in their offspring. Jap. J. Genet. 51, 161–174

    Google Scholar 

  • Orton, T.J. (1980): Chromosomal variability in tissue cultures and regenerated plants of Hordeum. Theor. Appl. Genet. 56, 101–112

    Google Scholar 

  • Rice, T.B.; Reid, R.K. Gordon, P.N. (1979): Morphogenesis in field crops. In: Propagation of Higher Plants Through Tissue Culture (eds.: Hughes, K.W.; Henke, R.; Constantin, M.), pp. 262–277. US Dept. of Energy, Technical Information Center

  • Sacristan, M.D.; Melchers, G. (1969): The caryological analysis of plants regenerated from tumorous and other callus cultures of tobacco. Mol. Gen. Genet. 105, 317–333

    Google Scholar 

  • Skoog, F.; Miller, C.O. (1957): Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp. Soc. Exp. Bio. 11, 118–131

    Google Scholar 

  • Smith, S.M.; Street, M.E. (1974): The decline of embryogenic potential as callus and suspension cultures of carrot (Daucus carota L.) are serially subcultured. Ann. Bot. 38, 223–241

    Google Scholar 

  • Snedecor, G.W.; Cochran, W.G. (1967): Statistical Methods. Ames: The Iowa State Univers. Press

    Google Scholar 

  • Torrey, J.G. (1967): Morphogenesis in relation to chromosomal constitution in long-term plant tissue cultures. Physiol. Plant. 20, 265–275

    Google Scholar 

  • Wenzel, G.; Schieder, O.; Prezewozny, T.; Sopory, S.K.; Melchers, G. (1979): Comparison of single cell culture derived Solanum tuberosum L. plants and a model for the application in breeding programs. Theor. Appl. Genet. 55, 49–55

    Google Scholar 

  • Wilcoxon, F.; Katti, S.K.; Wilcox, R.A. (1963): Critical values and probability levels for the Wilcoxon Rank Sum Test and the Wilcoxon Signed Rank test. New York: American Cyanamid Company; Tallahassee: Florida State Univers.

    Google Scholar 

  • Zagorska, N.A.; Shamina, Z.B.; Butenko, R.G. (1974): The relationship of morphogenetic potency of tobacco tissue culture and its cytogenetic features. Biol. Plant. 16, 262–274

    Google Scholar 

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Communicated by D. von Wettstein

Paper No. 6265 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina

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Traynor, P.L., Flashman, S.M. Hormone-induced caulogenesis in long-term tobacco cell lines and its effect on nuclear DNA content. Theoret. Appl. Genetics 60, 31–36 (1981). https://doi.org/10.1007/BF00275174

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  • DOI: https://doi.org/10.1007/BF00275174

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