Isolation and uptake of plant nuclei

  • Praveen K. Saxena
  • John King


Genetic transformation of animal cells by transplantation of isolated organelles, nuclei, and chromosomes is well documented and has played a significant role in investigating chromosome mapping and the regulation of gene expression [11, 13]. In plants, direct DNA transfer using isolated organelles was attempted following the discovery of techniques to eliminate the plant-specific barrier, the cell wall, which previously hampered the introduction of organelles into cells. Predictably, as soon as plant cells succumbed to procedures capable of enzymic degradation of cell walls, a whole new concept of handling naked plant cells (protoplasts) emerged as well as a genuine hope of being able to transform plant cells by foreign DNA introduction [15]. Thus, many workers studied the uptake of a variety of macromolecules and demonstrated the ability of plant protoplasts to accept foreign particles such as ferritin, bacteria, and isolated organelles like nuclei and chloroplasts (see references 7, 14, 21 for extensive reviews).


Nuclear Transplantation Plant Protoplast Somatic Cell Genetic Nuclear Uptake Ethylene Diamine Tetracetic Acid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Anderson JW (1986) Extraction of enzymes and subcellular organelles from plant tissues. Phytochemistry 7: 1973–1988.CrossRefGoogle Scholar
  2. 2.
    Becher D, Conrad B, Bottcher F (1982) Genetic transfer mediated by isolated nuclei in Saccharomyces cerevisiae. Current Genetics 6: 163–165.CrossRefGoogle Scholar
  3. 3.
    Dunham VL, Bryant JA (1983) Nuclei. In: Hall JZ, Moore AL (eds) Isolation of membranes and organelles from plant cells, pp 237–275. New York: Academic Press.Google Scholar
  4. 4.
    Ferenczy L, Pesti M (1982) Transfer of isolated nuclei into protoplasts of Saccharomyces cerevisiae. Curr Microbiol 7: 157–160.CrossRefGoogle Scholar
  5. 5.
    Fournier REK (1982) Microcell-mediated chromosone transfer. In: Shay JW (ed) Techniques in somatic cell genetics, pp 309–327. New York London: Plenum Press.CrossRefGoogle Scholar
  6. 6.
    Fowke LC (1986) Ultrastructural cytology of cultured plant tissues, cells, and protoplasts. In: Vasil IK (ed) Cell culture and somatic cell genetics of plants, pp 323–342. New York: Academic Press.Google Scholar
  7. 7.
    Fowke LC, Gamborg OL (1980) Applications of protoplasts to the study of plant cells. Int Rev Cytol 68: 9–51.CrossRefGoogle Scholar
  8. 8.
    Griesbach RJ (1987) Chromosome-mediated transformation via microinjection. Plant Sci 50: 69–77.CrossRefGoogle Scholar
  9. 9.
    Hadlaczky G, Bisztray G, Praznovszky T, Dudits D (1983) Mass isolation of plant chromosomes and nuclei. Planta 157: 278–285.CrossRefGoogle Scholar
  10. 10.
    Hughes BG, Hess WM, Smith MA (1977) Ultrastructure of nuclei isolated from plant protoplasts. Protoplasma 93: 267–274.CrossRefGoogle Scholar
  11. 11.
    Kao FT (1983) Somatic cell genetics and gene mapping. Int Rev Cytol 85: 109–146.PubMedCrossRefGoogle Scholar
  12. 12.
    Kobza J, Edwards GE (1984) Isolation of organelles: Chloroplasts. In: Vasil IK (ed) Cell culture and somatic cell genetics of plants, Vol 1, pp 471–482. New York: Academic Press.Google Scholar
  13. 13.
    Kondorosi E, Duda E (1980) Introduction of foreign genetic material into cultured mammalian cells by liposomes loaded with isolated nuclei. FEBS Lett 120: 37–40.PubMedCrossRefGoogle Scholar
  14. 14.
    Lörz H (1985) Isolated cell organelles and subprotoplasts — their role in somatic cell genetics. In: Dodds JH (ed) Plant genetic engineering, pp 27–59. London: Cambridge University Press.Google Scholar
  15. 15.
    Lörz H, Potrykus I (1976) Uptake of nuclei into higher plant protoplasts. In: Dudits D, Farkas GL, Maliga P (eds) Uptake of nuclei into higher plant protoplasts, pp 239–244. Budapest: Akadémiai Kiadó.Google Scholar
  16. 16.
    Lörz H, Potrykus I (1978) Investigations on the transfer of isolated nuclei into plant protoplasts. Theor Appl Genet 53: 251–256.Google Scholar
  17. 17.
    Melchers G, Labib G (1974) Somatic hybridization of plants by fusion of protoplasts. I. Selection of light resistant hybrids of “haploid” light sensitive varieties of tobacco. Mol Gen Genet 135: 277–294.CrossRefGoogle Scholar
  18. 18.
    Mii M, Saxena PK, Fowke LC, King J (1987) Isolation of chromosomes from cell suspension cultures of Vicia hajastana Grossh. Cytologia 52: 523–528.CrossRefGoogle Scholar
  19. 19.
    Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15: 473–497.CrossRefGoogle Scholar
  20. 20.
    Potrykus I, Hoffmann F (1973) Transplantation of nuclei into protoplasts of higher plants. Z Pflanzenphysiol 69: 287–289.CrossRefGoogle Scholar
  21. 21.
    Saxena PK, King J (1989) Isolation of nuclei and their transplantation into plant protoplasts. In: Bajaj YPS (ed) Biotechnology in Agriculture and Forestry, Vol 9, pp 328–342. Berlin Heidelberg: Springer-Verlag.Google Scholar
  22. 22.
    Saxena PK, Fowke LC, King J (1985a) An efficient procedure for isolation of nuclei from plant protoplasts. Protoplasma 128: 184–189.CrossRefGoogle Scholar
  23. 23.
    Saxena PK, Liu Y, Mii M, Fowke LC, King J (1985b) High nuclear yields from protoplasts of several plants. J. Plant Physiol 121: 193–197.CrossRefGoogle Scholar
  24. 24.
    Saxena PK, Mii M, Crosby WR, Fowke LC, King J (1986) Transplantation of isolated nuclei into plant protoplasts — A novel technique for introducing foreign DNA into plant cells. Planta 168: 29–35.CrossRefGoogle Scholar
  25. 25.
    Saxena PK, Liu Y, King J (1987) Nuclear transplantation into protoplasts: Optimal conditions for induction and determination of nuclear uptake. J Plant Physiol 128: 451–460.CrossRefGoogle Scholar
  26. 26.
    Sree Ramulu K, Verhoeven HA, Dijkhuis P (1988) Mitotic dynamics of micronuclei induced by amiprophos-methyl and prospects from chromosome-mediated gene transfer in plants. Theor Appl Genet 75: 575–584.CrossRefGoogle Scholar
  27. 27.
    Szabados L, Hadlaczky G, Dudits D (1981) Uptake of isolated plant chromosomes by plant protoplasts. Planta 151: 141–145.CrossRefGoogle Scholar
  28. 28.
    Tallman G, Reeck GR (1980) Isolation of nuclei from plant protoplasts without the use of a detergent. Plant Sci Lett 18: 271–275.CrossRefGoogle Scholar
  29. 29.
    Willmitzer L, Wagner KG (1981) The isolation of nuclei from tissue-cultured plant cells. Exp Cell Res 135: 69–77.PubMedCrossRefGoogle Scholar
  30. 30.
    Xiao W, Saxena PK, King J, Rank GH (1987) A transient duplication of the acetolactate synthase gene in a cell culture of Datura innoxia. Theor Appl Genet 74: 417–422.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 1992

Authors and Affiliations

  • Praveen K. Saxena
    • 1
  • John King
    • 2
  1. 1.Department of Horticultural ScienceUniversity of GuelphGuelphCanada
  2. 2.Department of BiologyUniversity of SaskatchewanSaskatoonCanada

Personalised recommendations