Summary
Somatic hybrid plants of Nicotiana tabacum (bar) (+) Nicotiana megalosiphon (npt II) were recovered after polyethylene glycol (PEG) mediated fusion. Hybrid calluses were selected on the basis of their dual resistance to bialaphos and kanamycin or UV inactivation of the donor species (Nicotiana megalosiphon) protoplasts. The hybrid nature of the individual clones obtained was confirmed by AFLP analysis. An array of plants were recovered including self-fertile hybrid plants with N. tabacum or N. megalosiphon phenotype, self-sterile plants with N. tabacum habit, leaf and intermediate flower morphology, self-sterile plants with N. megalosiphon habit, abnormal leaves and intermediate flowers, and self sterile plants of N. megalosiphon type with abnormal characters. Viable pollen was observed in hybrid plants from the third group. The hybrids possessed a nuclear DNA content near that of the diploid tobacco or N. megalosiphon, and also near that of the tetraploid genome size of N. megalosiphon. The results provide evidence for nonpreferential loss of one of the parental genomes and spontaneous asymmetrization of hybrid plants. The present study shows that by means of somatic hybridization a great genetic diversity in the hybrid clones can be achieved.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Bajaj, Y. P. S. Somatic hybrilization—a rich source of genetic variability. In: Bajaj, Y. P. S., ed. Biotechnology in agriculture and forestry, vol. 27. New York: Springer Verlag; 1994:3–32.
De Vries, S. E.; Ferwerda, M. A.; Leonen, A. E. H. M.; Pijnacker L. P.; Feenstra W. J. Chromosomes in somatic hybrids between Nicotiana plumbaginifolia and monoploid potato. Theor. Appl. Genet. 75: 170–176; 1987.
Dominique, M.; Spencer, B. C. A cytometric exercise in plant DNA histograms with 2C values for 70 species. Biol Cell. 78:41–45; 1993.
Donaldson, P.; Sproule, A.; Bevis, E.; Pandeya, R.; Keller, W.A.; Gleddie, S. Non-random chloroplast segregation in N. tabacum (+) N. rustica somatic hybrids selected by dual nuclear-encoded resistance. Theor. Appl. Genet. 86:465–473; 1993.
Famelaer, I.; Gleba, Y. Y.; Sidorov, V. A.; Kaleda, V. A.; Parokonny, A. S.; Boryshuk, N. V.; Cherep, N. N.; Negrutiu, I.; Jacobs, M. Intergeneric asymmetric hybrids between Nicotiana plumbaginifolia and Nicotiana sylvestris obtained by “gamma-fusion”. Plant Sci. 61:105–117; 1989.
Gleba, Y. Y.; Shlumukov, L. R. Somatic hybridization and cybridization. In: Bhojwani, S. S. ed. Development of crop science 19. Plant tissue culture: applications and limitations. Amsterdam: Elsevier; 1990:316–344.
Heller, R. Recherches sur la nutrition minérale des tissus végétaux cultivés in vitro. Ann. Sci. Nat. Bot. Biol. Végét. 14:1–223; 1953.
Hinnisdaels, S.; Bariller, L.; Mouras, A.; Sidorov, V.; Del-Favero, J.; Veuskens, J.; Negrutiu, I.; Jacobs, M. Highly asymmetric intergeneric nuclear hybrids between Nicotiana and Petunia: evidence for recombinations and translocation events in somatic hybrid plants after “gamma”-fusion N. tabacum. Theor. Appl. Genet. 82:609–614; 1991a.
Hinnisdaels, S.; Charti-Chhetri, G. B.; Negritiu, I. Somatic hybridization. Protoplast fusion—the PEG system. In: Negritiu, I.; Charti-Chhetri, G. B., eds. BioMethods. A laboratory guide for cellular and molecular plant biology, vol. 4. Basel, Boston, Berlin: Birkhäuser Verlag; 1991b:32–39.
Hinnisdaels, S.; Mouras, A.; Salesses, G.; Veuskens, J.; Taylor, C.; Charti-Chetri, G. B.; Negrutiu, I.; Jacobs, M. Translation events demonstrated by molecular, in situ hybridization and chromosome pairing analyses in highly asymmetric somatic hybrid plants. Transgenic Res. 1:170–176; 1992.
Hranov, A. Some problems on crossability between N. tabacum L. and N. megalosiphon Heurk et Müll. Bulgarian Tobacco 10:45–48; 1970.
Manolov, A. Transfer of complex resistance to blue mold and powdery mildew. Bulgarian Tobacco 12:9–17; 1981.
Morel, G. Sur la developpement de tissus de vignes cultivees in vitro. C. R. Acad. Sci. Soc. Biol. Paris. 138:62; 1944.
Murashige, T.; Skoog, F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.
Palakarcheva, M. Interspecific hybridization in g. Nicotiana. Sofia: Bulgarian Academy of Sciences Publishing House; 1988.
Pelletier, G. Somatic hybridization. In: Hayward, M. D.; Bosemark, N. O.; Romagosa, I., eds. Plant breeding: principles and prospects. London: Chapman & Hall; 1993:93–106.
Piastuch, W. C.; Bates, G. W. Chromosomal analysis of Nicotiana asymmetric somatic hybrids by dot blotting and in situ hybridisation. Mol. Gen. Genet. 222:97–103; 1990.
Sproule, A.; Donaldson, P.; Dijak, M.; Bevis, E.; Pandeya, R.; Keller, W. A.; Gleddie, S. Fertile somatic hybrids between transgenic Nicotiana tabacum and transgenic Nicotiana debneyi selected by dual antibiotic resistance. Theor. Appl. Genet. 82:450–456; 1991.
Stavely, J. R. Disease resistance. Nicotiana: Proceeding for experimental use. U.S. Department of Agriculture 1:87–110; 1979.
Vos, P.; Hogers, R.; Bleeker, M.; Reijans, M.; van de Lee, T.; Hornes, M.; Frijters, A.; Pot, J.; Peleman, J.; Kupier, M.; Zabeau, M. AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res. 25:4407–4414; 1995.
Zelcer, A.; Aviv, D.; Galun, E. Interspecific transfer of cytoplasmic male sterility by fusion between protoplasts of normal Nicotiana sylvestris and X-irradiated protoplasts of male sterile N. tabacum. Z. Pflanzenphysiol. 90:397–407; 1978.
Zhu, X.Y.; Negrutiu, I. Isolation and culture of protoplasts. In: Negritiu, I.; Charti-Chhetri, G. B., eds. BioMethods. A laboratory guide for cellular and molecular plant biology. Basel, Boston, Berlin: Birkhäuser Verlag; 1991:18–27.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ilcheva, V., San, L.H., Dimitrov, B. et al. Morphological and cytological characteristics of somatic hybrids of Nicotiana tabacum L. (+) N. megalosiphon Heurk. et Müll. In Vitro Cell.Dev.Biol.-Plant 36, 69–73 (2000). https://doi.org/10.1007/s11627-000-0016-4
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11627-000-0016-4