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Somatic hybridization between male sterile Nicotiana tabacum and N. glutinosa through protoplast fusion

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Summary

Protoplasts derived from suspension cultured cells of cytoplasmic male sterile Nicotiana tabacum (N. debneyi cytoplasm) and of fertile N. glutinosa were fused with the aid of polyethylene glycol (PEG). Out of 1,089 colonies developed from PEG-treated protoplasts, 29 restored whole plants.

A somatic hybrid plant was selected on the basis of isoelectrofocusing analysis of Fraction I protein in leaves of regenerated plants. A newly created hybrid contained small subunits of both parents but only a N. glutinosa type large subunit.

Male sterile character was conserved in a hybrid plant while leaf morphology was intermediate between the parents. By tobacco mosaic virus infection tests, the hybrid's leaves showed resistant symptoms, hypersensitive local lesions, which were due to N. glutinosa nuclear genome expression.

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Abbreviations

PEG:

Polyethylene glycol

TMV:

Tobacco mosaic virus

Literature

  1. Aviv, D.; Fluhr, R.; Edelman, M.; Galun, E.; (1980): Progeny analysis of the interspecific somatic hybrids: Nicotiana tabacum (CMS) + Nicotiana sylvestris with respect to nuclear and chloroplast markers. Theor. Appl. Genet. 56, 145–150

  2. Belliard, G.; Vedel, F.; Pelletier, G. (1979): Mitochondrial recombination in cytoplasmic hybrids of Nicotiana tabacum by protoplast fusion. Nature 281, 401–403

  3. Belliard, G.; Pelletier, G.; Vedel, F.; Quetier, F. (1978): Morphological characteristics and chloroplast DNA distribution in different cytoplasmic parasexual hybrids of Nicotiana tabacum. Molec. Gen. Genet. 165, 231–237

  4. Chan, P.H.; Wildman, S.G. (1972): Chloroplast DNA codes for the primary structure of the large subunit of Fraction I protein. Biochim. Biophys. Acta 277, 677–680

  5. Chen, K.; Johal, S.; Wildman, S.G. (1976): Phenotypic markers for chloroplast DNA genes in higher plants and their use in biochemical genetics. In: Nucleic Acids and Protein Synthesis in Plants (eds. Borgrad, L.; Weil, J.H.), pp. 183–194. New York: Plenum Press

  6. Chen, K.; Wildman, S.G.; Smith, H.H. (1977): Chloroplast DNA distribution in parasexual hybrids as shown by polypeptide composition of Fraction I protein. Proc. Natl. Acad. Sci. (USA) 74, 5109–5112

  7. Clausen, R.E.; Cameron, D.R. (1957): Inheritance in Nicotiana tabacum. XXVIII. The cytogenetics of introgression. Proc. Natl. Acad. Sci. (USA) 43, 908–913

  8. Evans, D.A.; Wetter, L.R.; Gamborg, O.L. (1980): Somatic hybrid plants of Nicotiana glauca and Nicotiana tabacum obtained by protoplast fusion. Physiol. Plant 48, 225–230

  9. Gleba, Y.Y.; Piven, N.M.; Komarnitskii, I.K.; Sytnik, A.K.M. (1978): Parasexual cytoplasmic Nicotiana tabacum + N. debneyi hybrids (cybrids) obtained by protoplast fusion. Doklady Akademii Nauk SSSR 240, 1223–1226

  10. Imamura, J.; Harada, H. (1980): Studies on the changes in the volume and proliferation rate of cells during embryogenesis of in vitro cultured pollen grains of Nicotiana tabacum L. Z. Pflanzenphysiol. 96, 261–267

  11. Izhar, S.; Power, J.B. (1979): Somatic hybridization in petunia: A male sterile cytoplasmic hybrid. Plant Sci. Lett. 14, 49–55

  12. Kao, K.N.; Michayluk, M.R. (1974): A method for high-frequency intergeneric fusion of plant protoplasts. Planta 115, 355–367

  13. Kung, S.D.; Gray, J.C.; Wildman, S.G.; Carlson, P.S. (1975): Polypeptide composition of Fraction I protein from parasexual hybrid plants in the genus Nicotiana. Science 187, 353–355

  14. Melchers, G.; Sacristan, M.D.; Holder, A.A. (1978): Somatic hybrid plants of potato and tomato regenerated from fused protoplasts. Carlsberg. Res. Commun. 43, 203–218

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

  16. Sand, S.A.; Christoff, G.T. (1973): Cytoplasmic-chromosomal interactions and altered differentiation in tobacco. J. Heredity 64, 24–30

  17. Uchimiya, H.; Murashige, T. (1974): Evaluation of parameters in the isolation of viable protoplasts from cultured tobacco cells. Plant Physiol. 54, 936–944

  18. Uchimiya, H.; Wildman, S.G. (1979): Nontranslation of foreign genetic information for Fraction I protein under circumstances favorable for direct transfer of Nicotiana gossei isolated chloroplasts into N. tabacum protoplasts. In Vitro 15, 463–468

  19. Uchimiya, H.; Chen, K.; Wildman, S.G. (1979) A micro electrofocusing method for determining the large and small subunit polypeptide composition of Fraction I proteins. Plant Sci. Lett. 14, 387–394

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Communicated by K. Tsunewaki

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Uchimiya, H. Somatic hybridization between male sterile Nicotiana tabacum and N. glutinosa through protoplast fusion. Theoret. Appl. Genetics 61, 69–72 (1982). https://doi.org/10.1007/BF00261514

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Key words

  • Protoplast fusion
  • Cytoplasmic male sterility
  • Tobacco
  • Fraction I protein
  • TMV resistance