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Variation in nuclear DNA content of isonicotinic acid hydrazide-resistant cell lines and mutant plants of Nicotiana tabacum

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Summary

Isonicotinic acid hydrazide (INH)-resistant lines of Nicotiana tabacum have been maintained in callus culture for six years and mutant plants have been regenerated from a number of these lines. This study examines variations in DNA content in nuclei of several of these callus cultures, regenerated plants, and secondary callus from the regenerated plants. The lines selected for study include three easily regenerated lines (I 21, I 24, and I 9) and two lines of poor regenerating capacity (I 1 and I 18). Two of the regenerating lines eventually led to fertile plants and the third produced only sterile plants. In general, the range of total nuclear variability was not as high as anticipated from other studies of long-term tobacco callus cultures. The majority of nuclei in all the distributions were between 3 and 20 pg, and the most frequently encountered distributions concentrated in the 7–18 pg region corresponding to 2–5C by our estimate of the C value for tobacco. Distributions were not identical for plants regenerated from the same culture simultaneously, and the nuclear DNA content of secondary callus cultures from one of the plants examined did not reflect the quantitative DNA pattern of the plant from which it was derived. The greatest degree of variability and highest DNA content for individual nuclei were observed in the primary callus of the poorly- and non-regenerating lines. The variability in DNA content was not associated with the INH-resistant trait.

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Literature

  • Bayliss, M.W. (1977): Factors affecting the frequency of tetraploid cells in a predominantly diploid suspension culture of Daucus carota. Protoplasma 92, 109–115

    Google Scholar 

  • Bayliss, M.W. (1980): Chromosomal variation in plant tissues in culture. In: Intern. Rev. of Cytology, Supplement 11A, Perspectives in Plant Cell and Tissue Culture (ed. Vasil, I.), pp. 113–144. New York: Acad. Press

    Google Scholar 

  • Bennici, A.; Buiatti, M.; D'Amato, F.; Pagliai, M. (1971): Nuclear behavior in Haplopappus gracilis callus grown in vitro on different culture media. In: Colloq. Intern. No. 193, Les Cultures de Tissus de Plantes, pp. 245–250. Paris: C.N.R.S.

    Google Scholar 

  • Berlyn, G.P.; Dhillon, S.S.; Miksche, J.P. (1979): Feulgen cytophotometry of pine nuclei, II. Effect of pectinase used in cell separation. Stain Technol. 54, 201–204

    Google Scholar 

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

    Google Scholar 

  • Berlyn, M.B. (1980): Isolation and characterization of isonicotinic acid hydrazide-resistant mutants of Nicotiana tabacum. Theor. Appl. Genet. 58, 19–26

    Google Scholar 

  • Binns, A.; Meins, F. (1980): Chromosome number and the degree of cytokinin habituation of cultured tobacco pith cells. Protoplasma 103, 179–187

    Google Scholar 

  • Brossard, D. (1975): La néoformation de bourgeons végétatifs a partir de la moelle du tabac (Nicotiana tabacum L., var. Wisconsin 38) cultivée in vitro. Analyse cytochimique, historadiographique et cytophotométrique. Ann. Sci. Nat. Bot. 16, 43–150

    Google Scholar 

  • D'Amato, F. (1972): Morphogenetic aspects of the development of meristems in seed embryos. In: The Dynamics of Meristem Cell Populations (eds. Miller, M.W.; Kuehnert, C.C.), pp. 149–163. New York: Plenum Press

    Google Scholar 

  • D'Amato, F. (1977): Cytogenetics of differentiation in tissue and cell cultures. In: Plant Cell, Tissue, and Organ Culture (eds. Reinert, J.; Bajaj, Y.), pp. 343–357. Berlin, Heidelberg, New York: Springer

    Google Scholar 

  • D'Amato, F. (1978): Chromosome number variation in cultured cells and regenerated plants. In: Frontiers of Plant Tissue Culture 1978 (ed. Thorpe, T.A.) Calgary: Intern. Assoc. for Plant Tissue Culture 1978

    Google Scholar 

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

    Google Scholar 

  • Fox, J.E. (1963): Growth factor requirements and chromosome number in tobacco tissue cultures. Physiol. Plant. 16, 793–794

    Google Scholar 

  • Ghosh, A.; Gadgil, V. (1979): Shift in ploidy level of callus tissue: a function of growth substances. Indian J. Exp. Biol. 17, 562–564

    Google Scholar 

  • Kibler, R.; Neumann, K.-H. (1980): On cytogenetic stability of cultured tissue and cell suspensions of haploid and diploid origin. In: Plant Cell Cultures: Results and Perspectives (eds. Sala, F.; Parisi, B.; Cella, R.; Ciferri, O.), pp. 59–65. New York: Elsevier

    Google Scholar 

  • Muir, W.H. (1965): Influence of variation in chromosome number on differentiation in plant tissue cultures. In: Proc. Intern. Conf. Plant Tissue Cultures (eds. White, P.R.; Grove, A.R.), pp. 485–492. Berkeley, McCutchan

    Google Scholar 

  • Murashige, T.; Nakano, R. (1967): Chromosome complement as a determinant of the morphogenic potential of tobacco cells. Am. J. Bot. 54, 963–970

    Google Scholar 

  • Nandi, S.; Fridborg, G.; Eriksson, T. (1977): Effects of 6-(3-methyl-2-buten-1-ylamino purine and α-naphthaleneacetic acid on root formation and cytology of root tips and callus in tissue cultures of Allium cepa var. ‘proliferum’. Hereditas 85, 57–62

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Partanen, C.R. (1965): Cytological behavior of plant tissues in vitro as a reflection of potentialities in vivo. In: Proc. Intern. Conference Plant Tissue Culture (eds. White, P.R.; Grove, A.R.), pp. 463–471. Berkeley: McCutchan

    Google Scholar 

  • Patel, K.R.; Berlyn, G.P. (1982): Genetic instability of multiple buds of Pinus coulteri regenerated from tissue culture. Can. J. For. Res. 12

  • Rasch, E.M.; Barr, H.J.; Rasch, R.W. (1971): The DNA content of sperm of Drosophila melanogaster. Chromosoma 33, 1–18

    Google Scholar 

  • Sacristan, M.D.; Lutz, A.M. (1970): Etude de nombre chromosomique sur des plants de tabac d'origine unicellulaire. C.R. Acad. Sci. (Paris) Ser. D. 270, 1334–1336

    Google Scholar 

  • 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 

  • Shimada, T.; Tabata, M. (1967): Chromosome numbers in cultured pith tissue of tobacco. Jpn. J. Genet. 42, 195–201

    Google Scholar 

  • Siegel, A.; Lightfoot, D.; Ward, O.G.; Keene, S. (1973): DNA complementary to ribosomal RNA: relation between genomic proportion and ploidy. Science 179, 682–683

    Google Scholar 

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

    Google Scholar 

  • Singh, B.D.; Harvey, B.L. (1975): Cytogenetic studies on Haplopappus gracilis cells cultured on agar and in liquid media. Cytologia 40, 347–354

    Google Scholar 

  • Torrey, J.G. (1961): Kinetin as trigger for mitosis in mature endomitotic plant cells. Exp. Cell Res. 23, 281–299

    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 

  • Traynor, P.L.; Flashman, S.M. (1981): Hormone-induced caulogenesis in long-term tobacco cell lines and its effect on nuclear DNA content. Theor. Appl. Genet. 60, 31–36

    Google Scholar 

  • van't Hof, J. (1974): Control of the cell cycle in higher plants. In: Cell Cycle Controls (eds. Padilla, G.M.; Cameron, I.L.; Zimmerman, A.), pp. 77–85. New York: Acad. Press

    Google Scholar 

  • van't Hof, J.; McMillan, B. (1969): Cell population kinetics in callus tissues of cultured pea root segments. Am. J. Bot. 56, 42–51

    Google Scholar 

  • Zagorska, N.A.; Shamino, 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 

  • Zelitch, I.; Berlyn, M.B. (1982): Altered glycine decarboxylation inhibition in isonicotinic acid hydrazide-resistant mutant callus lines and in regenerated plants and seed progeny. Plant Physiol. 69

  • Zimmerman, J.L.; Goldberg, R.B. (1977): DNA sequence organization in the genome of Nicotiana tabacum, Chromosoma 59, 227–252

    Google Scholar 

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Communicated by F. Mechelke

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Berlyn, M.B. Variation in nuclear DNA content of isonicotinic acid hydrazide-resistant cell lines and mutant plants of Nicotiana tabacum . Theoret. Appl. Genetics 63, 57–63 (1982). https://doi.org/10.1007/BF00303491

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  • DOI: https://doi.org/10.1007/BF00303491

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