Summary
‘Tabac anergié’ tissue has been used by many investigators as a habituated tobacco cell line. In this study, we have found sequences which are homologous to approximately 9 megadaltons of an Agrobacterium octopine-type plasmid in DNA isolated from ‘Tabac anergié’ tissue. These T-DNA sequences were shown to be the same ‘core DNA’ sequences found to be present in several other unorganized tumor lines which have been studied in our laboratory. The T-DNA was also shown to be integrated into plant DNA. Thus, the ‘Tabac anergié’ tissue is a crown gall tumor induced by a strain of Agrobacterium tumefaciens containing an octopinetype plasmid. These results indicated that the plants regenerated from 40 clones of the ‘Tabac anergié’ line by Sacristán and Melchers (1977) were actually reverted plants obtained from an unorganized crown gall tumor, kept in culture for more than 25 years. It is necessary to re-evaluate observations and conclusions which have been based upon ‘Tabac anergié’ being a habituated tissue.
References
Blair, D.G., Sherratt, D.J., Clewell, D.B., Helinski, D.R.: Isolation of supercoiled colicinogenic factor E1 DNA sensititive to ribonuclease and alkali. Proc. Natl. Acad. Sci. USA 69, 2518 (1972)
Bomhoff, G.H.: Studies on Crown gall — a plant tumor. Investigation on protein composition and on the use of guanidine compounds as a marker for transformed cells. Thesis, University of Leiden, The Netherlands (1974)
Bomhoff, G., Klapwijk, P.M., Kester, H.C.M., Schilperoot, R.A., Hernalsteens, J.P., Schell, J.: Octopine and nopaline synthesis and breakdown genetically controlled by a plasmid of Agrobacterium tumefaciens. Mol. Gen. Genet. 145, 177 (1976)
Buitti, M., Bennici, A.: Callus formation and habituation in Nicotiana species in relation to the specific ability for dedifferentiation. Lincei-Rend. Sci. Fis. Mat. Nat. 48, 261 (1970)
Braun, A.C., Wood, H.N.: Suppression of the neoplastic state with the acquisition of specialized functions in cells, tissues, and organs of crown gall teratoma of tobacco. Proc. Natl. Acad. Sci. USA 73, 496 (1976)
Chilton, M.D.: Ribosomal DNA in a nuclear satellite of tomato. Genetics 81, 469 (1975)
Chilton, M.D., Drummond, M.H., Merlo, D.J., Sciaky, D., Montoya, A.L., Gordon, M.P., Nester, E.W.: Stable incorporation of plasmid DNA into higher plant cells: The molecular basis of crown gall tumorigenesis. Cell 11, 263 (1977)
Dons, J.J.M., Valentijn, M., Schilperoot, R.A., Duijn, P. van: Nuclear DNA content and phytohormone requirements of normal crown gall and habituated tissues of Nicotiana tabacum var. White Burley. Exp. Cell Res. 89, 283 (1974)
Gautheret, R.J.: Hétéroanxines et cultures de tissue végétaux. Bull. Soc. Chim. Biol. 24, 13 (1942)
Gautheret, R.J.: Nutrition of plant tissue cultures. Ann. Rev. Plant Physiol. 6, 433–484 (1955)
Heyn, R.F., Hermans, A.K., Schilperoot, R.A.: Rapid and efficient isolation of highly polymerized plant DNA. Plant Sci. Lett. 2, 73 (1973)
Linsmaier, E.M., Skoog, F.: Organic growth factor requirements of tobacco tissue cultures. Physiol. Plant. Pathol. 18, 100 (1965)
Lutz, A.: Obtention de plantes de Tabac a partir de cultures unicellularies provenant d'une souche anergiée. C.R. Acad. Sci. (Paris) 262, 1856 (1966)
Maniatis, T., Jeffrey, A., Kleid, D.G.: Nucleotide sequence of the rightward operator of phage λ. Proc. Natl. Acad. Sci. USA 72, 1184 (1975)
Meins, F.: Reversal of the Neoplastic state in plants. Am. J. Pathol. 89, 687 (1977)
Meins, F., Binns, A.: Epigenetic variation of cultured somatic cells: Evidence for gradual changes in the requirement for factors promoting cell division. Proc. Natl. Acad. Sci. USA 74, 2928 (1977)
Merlo, D.J., Nutter, R.C., Montoya, A.L., Garfinkel, D.J., Drummond, M.H., Chilton, M.-D., Gordon, M.P., Nester, E.W.: The boundaries and copy numbers of Ti-plasmid T-DNA vary in crown gall tumors. (sumitted)
Morel, G.: Recherches sur la culture associée de parasites obligatories et de tissues végétaux. Thesis, Paris, in: Ann. Epiphyties 14, mémoire no. 5 (1948)
Sacristán, M., Melchers, G.: Regeneration of plants from “Habituated” and “Agrobacterium transformed” single-cell clones of tobacco. Mol. Gen. Genet. 152, 111 (1977)
Sheridan, W.F.: Tissue culture of the monocot Lilium. Planta 82, 189 (1968)
Southern, E.M.: Detection of specific sequence among DNA fragments separated by gel, electrophoresis. J. Mol. Biol. 98, 503 (1975)
Syono, K., Furuya, T.: Induction of Auxin-Nonrequiring tobacco calluses and its reversal by treatments with auxin. Plant and Cell Physiol. 15, 7 (1974)
Thomashow, M.F., Nutter, R., Montoya, A.L., Gordon, M.P., Nester, E.W.: Integration and organization of Ti-plasmid sequences in Crown Gall tumors. Cell, in press (1980)
Wendt,-Gallitelli, M.F., Dobrigkeit, I.: Investigations implying the invalidity of octopine as a marker for transformation by Agrobacterium tumefaciens. Z. Naturforsch. 28c, 768 (1973)
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Yang, F., Merlo, D.J., Gordon, M.P. et al. Plasmid DNA of Agrobacterium tumefaciens detected in a presumed habituated tobacco cell line. Molec. Gen. Genet. 179, 223–226 (1980). https://doi.org/10.1007/BF00268467
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00268467