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Theoretical and Applied Genetics

, Volume 72, Issue 1, pp 48–52 | Cite as

The production of fertile, triploid somatic hybrid plants (Nicotiana glutinosa (n) + N. tabacum (2n)) via gametic: somatic protoplast fusion

  • A. Pirrie
  • J. B. Power
Originals

Summary

The fusion of gametic protoplasts with somatic protoplasts giving rise to gametosomatic hybrid plants was investigated. Gametosomatic hybrid plants were regenerated following the fusion of nitrate reductase deficient (Nr) Nicotiana tabacum Nia-130 leaf mesophyll protoplasts with N. glutinosa tetrad protoplasts. The resulting plants were confirmed as hybrids, based on leaf and floral morphology, chromosome number, leaf esterase and leaf callus peroxidase zymograms and Fraction-1-protein analysis. The five gametosomatic hybrid plants had the expected pentaploid, but functionally triploid chromosome number of 3n=5x=60. The relevance of triploid gametosomatic hybrids in facilitating limited gene transfer, is discussed. The utilisation of tetrads as a generally available source of haploid protoplasts for fusion studies is proposed.

Key words

Nicotiana Gametosomatic hybridisation Tetrad protoplasts Triploids 

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References

  1. Bajaj YPS (1974) Isolation and culture studies on pollen and pollen mother cell-protoplasts. Plant Sci Lett 3:93–99Google Scholar
  2. Bhojwani SS, Cocking EC (1972) Isolation of protoplasts from pollen tetrads. Nature New Biol 239:29–30Google Scholar
  3. Cammearts D, Jacobs M (1980) A simple electrophoretic procedure for the determination of the polypeptide composition of the subunits of Fraction I protein. Anal Biochem 97:317–320Google Scholar
  4. Cooper-Bland S, Pental D, Cocking EC (1985) Somatic hybridisation between nitrate reductase deficient N. tabacum and N. glutinosa. (in prep)Google Scholar
  5. Deka PC, Mehra AK, Pathak NN, Seu SK (1977) Isolation and fusion studies on protoplasts from pollen tetrads. Experientia 33:182–184PubMedGoogle Scholar
  6. Dudits D, Hadlaczky GY, Bajszar GY, Koncz CS, Lazar G, Horvath G (1979) Plant regeneration from intergeneric cell hybrids. Plant Sci Lett 15:101–112Google Scholar
  7. Dudits D, Fejer O, Hadlaczky GY, Koncz CS, Lazar GB, Horvath G (1980) Intergeneric gene transfer mediated by plant protoplast fusion. Mol Gen Genet 179:283–288Google Scholar
  8. Frearson EM, Power JB, Cocking EC (1973) The isolation, culture and regeneration of Petunia leaf protoplasts. Dev Biol 33:130–137Google Scholar
  9. Hamill JD, Pental D, Cocking EC, Muller AJ (1983) Production of a nitrate reductase deficient streptomycin resistant mutant of Nicotiana tabacum for somatic hybridisation studies. Heredity 50:197–200Google Scholar
  10. Keller WA, Melchers G (1973) The effect of high pH and calcium on tobacco leaf protoplast fusion. Z Naturforsch 28c:737–741Google Scholar
  11. Maheshwari SC, Tyagi AK, Malhotra K, Sopary SK (1980) Induction of haploidy from pollen grains in Angiosperms — the current status. Theor Appl Genet 58:193–206Google Scholar
  12. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479Google Scholar
  13. Nagao T (1979) Somatic hybridisation of protoplasts. 2. The combinations of Nicotiana tabacum and N. glutinosa and N. tabacum and N. alata. Jpn J Crop Sci 48(3): 385–392Google Scholar
  14. Negrutiu I (1980) Nutritional requirements of protoplast-derived haploid tobacco cells grown at low cell densities in liquid medium. Planta 149:7–18Google Scholar
  15. Pental D, Cooper-Bland S, Harding K, Cocking EC, Muller AJ (1982) Cultural studies on nitrate reductase deficient Nicotiana tabacum mutant protoplasts. Z Pflanzenphysiol 105:219–227Google Scholar
  16. Pental D, Cocking EC (1985) Some theoretical and practical possibilities of plant genetic manipulation using protoplasts. Hereditas (Suppl) 3:83–92Google Scholar
  17. Power JB, Frearson EM, Hayward C, Cocking EC (1975) Some consequences on the fusion and selective culture of Petunia and Parthenocissus protoplasts. Plant Sci Lett 5: 197–207Google Scholar
  18. Smith HH, Hamill DE, Weaver EA, Thompson KH (1970) Multiple molecular forms of peroxidases and esterases among Nicotiana species and amphiploids. J Hered 61: 203–212Google Scholar
  19. Uchimaya H (1982) Somatic hybridisation between male sterile Nicotiana tabacum and N. glutinosa through protoplast fusion. Theor Appl Genet 61:69–72Google Scholar
  20. Uchimaya H, Kobayashi S, Ono M, Brar DS, Harada N (1984) Characterisation of nuclear and cytoplasmic information in the progeny of a somatic hybrid between male sterile Nicotiana tabacum and N. glutinosa. Theor Appl Genet 68: 95–100Google Scholar

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • A. Pirrie
    • 1
  • J. B. Power
    • 1
  1. 1.Plant Genetic Manipulation Group, Department of BotanyUniversity of NottinghamNottinghamUK

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