Advertisement

Electrofusion: The Technique and Its Application to Somatic Hybridization

  • G. W. Bates
Part of the Biotechnology in Agriculture and Forestry book series (AGRICULTURE, volume 9)

Abstract

The use of electric fields for inducing protoplast fusion (electrofusion) has its origins in biophysical studies of cell membranes. However, recent work showing electrofusion to be simpler, quicker, and often more efficient than chemically induced fusion is attracting the interest of geneticists and tissue-culture practitioners. Indeed, electrofusion may become the fusion technique of choice for species whose cells exhibit a severe cytotoxic response to polyethylene glycol (PEG). Interest in this area has been further heightened by recent demonstrations that electric pulses can be used to introduce foreign DNA into plant cells (“electroporation”: Fromm et al. 1986; Riggs and Bates 1986). This chapter provides an overview of electrofusion and its application to plant somatic hybridization.

Keywords

Somatic Hybridization Fusion Product Mesophyll Protoplast Plant Protoplast Protoplast Suspension 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bates GW (1985) Electrical fusion for optimal formation of protoplast heterokaryons in Nicotiana. Planta 165: 217–224CrossRefGoogle Scholar
  2. Bates GW, Gaynor JJ, Shekhawat NS (1983) Fusion of plant protoplasts by electric fields. Plant Physiol 72: 1110–1113PubMedCrossRefGoogle Scholar
  3. Bates GW, Hasenkampf CA (1985) Culture of plant somatic hybrids following electrical fusion. Theor Appl Genet 70: 227–233CrossRefGoogle Scholar
  4. Bates GW, Hasenkampf CA, Contolini CS, Piastuch WC (1987a) Limited gene transfer by somatic cell fusion in Nicotiana. Theor Appl Genet 74: 718–726CrossRefGoogle Scholar
  5. Bates GW, Nea LJ, Hasenkampf CA ( 1987 b) Electrofusion and plant somatic hybridization. In: Sowers AE (ed) Cell fusion. Plenum, New York, pp 479–496Google Scholar
  6. Bates GW, Saunders JA, Sowers AE (1987c) Electrofusion, principles and applications. In: Sowers AE (ed) Cell fusion. Plenum, New York, pp 367–395Google Scholar
  7. Chapel M, Teissie J, Alibert G (1984) Electrofusion of spermine-treated plant protoplasts. FEBS Lett 173: 331–336CrossRefGoogle Scholar
  8. Chapel M, Montane M-H, Ranty B, Teissie J, Alibert G (1986) Viable somatic hybrids are obtained by direct current electrofusion of chemically aggregated plant protoplasts. FEBS Lett 196: 79–86CrossRefGoogle Scholar
  9. Constabel F, Dudits D, Gamborg OL, Kao KN (1975) Nuclear fusion in intergeneric heterokaryons. A note. Can J Bot 53: 2092–2095CrossRefGoogle Scholar
  10. de Vries SE, Tempelaar MJ (1987) Electrofusion and analysis of potato somatic hybrids. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry 3. Potato. Springer, Berlin Heidelberg New York Tokyo, pp 211–222Google Scholar
  11. Fromm ME, Taylor LP, Walbot V (1986) Stable transformation of maize after gene transfer by electroporation. Nature 319: 791–793PubMedCrossRefGoogle Scholar
  12. Ghose TK, Golds TJ, Davey MR, Cocking EC (1986) An assessment of electrically and chemically induced protoplast fusion in forage legumes. Poster 434, VI Int Congr Plant tissue and cell culture, August 3–8, Univ Minnesota, Minneapolis, p 385Google Scholar
  13. Kao KN (1975) A method for fusion of plant protoplasts with polyethylene glycol. In: Gamborg OL, Wetter LR (eds) Plant tissue culture methods. Prairie Research Laboratory, Saskatoon, Sask., Can, pp 22–27Google Scholar
  14. Kinosita K, Tsong TY (1977) Formation and resealing of pores of controlled sizes in human erythrocyte membrane. Nature 268: 438–441PubMedCrossRefGoogle Scholar
  15. Kinosita K, Tsong TY (1979) Voltage-induced conductance in human erythrocyte membranes. Biochim Biophys Acta 554: 479–497PubMedCrossRefGoogle Scholar
  16. Kohn H, Schieder HR, Schieder O (1985) Somatic hybrids in tobacco mediated by electrofusion. Plant Sci 38: 121–128CrossRefGoogle Scholar
  17. Koop H-U, Schweiger H-G (1985) Regeneration of plants after electrofusion of selected pairs of protoplasts. Eur J Cell Biol 39: 46–49Google Scholar
  18. Mehrle W, Zimmermann U, Hampp R (1985) Evidence for asymmetrical uptake of fluorescent dyes through electro-permeabilized membranes of Avena mesophyll protoplasts. FEBS Lett 185: 89–94CrossRefGoogle Scholar
  19. Morikawa H, Sugino K, Hayashi Y, Takeda J, Senda M, Hirai A, Yamada Y (1986) Interspecific plant hybridization by electrofusion in Nicotiana. Biotechnology 4: 57–60CrossRefGoogle Scholar
  20. Naton B, Mehrle W, Hampp R, Zimmermann U (1986) Mass electrofusion and mass selection of functional hybrids from vacuolate x evacuolate protoplasts. Plant Cell Rep 5: 419–422CrossRefGoogle Scholar
  21. Nea LJ, Bates GW (1987) Protoplast electrofusion: factors affecting fusion efficiency and cell viability. Plant Cell Rep 6: 337–340CrossRefGoogle Scholar
  22. Nea LJ, Bates GW, Gilmer PJ (1987) Facilitation of electrofusion of plant protoplasts by membrane active agents. Biochim Biophys Acta 897: 293–301CrossRefGoogle Scholar
  23. Negrutiu I, De Brouwer D, Watts JW, Sidorov VI, Dirks R, Jacobs M (1986) Fusion of plant protoplasts: a study using auxotrophic mutants of Nicotiana plumbaginifolia, Viviani. Theor Appl Genet 72: 279–286CrossRefGoogle Scholar
  24. Pohl HA (1978) Dielectrophoresis. Cambridge Univ Press, LondonGoogle Scholar
  25. Puite KJ, van Wikselaar P, Verhoeven H (1985) Electrofusion, a simple and reproducible technique in somatic hybridization of Nicotiana plumbaginifolia mutants. Plant Cell Rep 4: 274–276CrossRefGoogle Scholar
  26. Puite KJ, Roest S, Pijnacker LP (1986) Somatic hybrid potato plants after electrofusion of diploid Solanum tuberosum and Solanum phureja. Plant Cell Rep 5: 262–265CrossRefGoogle Scholar
  27. Riggs CD, Bates GW (1986) Stable transformation of tobacco by electroporation: evidence for plasmid concatenation. Proc Natl Acad Sci USA 83: 5602–5606PubMedCrossRefGoogle Scholar
  28. Ruzin SE, McCarthy SC (1986) The effect of chemical facilitators on the frequency of electrofusion of tobacco mesophyll protoplast. Plant Cell Rep 5: 342–345CrossRefGoogle Scholar
  29. Scheurich P, Zimmermann U (1981) Electrically stimulated fusion of different plant cell protoplasts. Plant Physiol 67: 849–853PubMedCrossRefGoogle Scholar
  30. Senda M, Takeda J, Abe S, Nakamura T (1979) Induction of cell fusion of plant protoplasts by electrical stimulation. Plant Cell Physiol 20: 1441–1443Google Scholar
  31. Spangenberg G, Schweiger H-G (1986) Controlled electrofusion of different types of protoplasts and subprotoplasts including cell reconstitution in Brassica napus L. Eur J Cell Biol 41: 51–56Google Scholar
  32. Tempelaar MJ, Jones MGK (1985 a) Fusion characteristics of plant protoplasts in electric fields. Planta 165: 205–216CrossRefGoogle Scholar
  33. Tempelaar MJ, Jones MGK (1985 b) Directed electrofusion between protoplasts with different responses in a mass fusion system. Plant Cell Rep 4: 92–95CrossRefGoogle Scholar
  34. Tempelaar MJ, Duyst A, De Vlas Sy, Krol G, Symmonds C, Jones MGK (1987) Modulation and direction of the electrofusion response in plant protoplasts. Plant Sci 48: 99–105CrossRefGoogle Scholar
  35. Vienken J, Ganser R, Hampp R, Zimmermann U (1981) Electric field-induced fusion of isolated vacuoles and protoplasts of different developmental and metabolic provenience. Physiol Plant 53: 64–70CrossRefGoogle Scholar
  36. Watts JW, King JM (1984) A simple method for large-scale electrofusion and culture of plant protoplasts. Biosci Rep 4: 335–342PubMedCrossRefGoogle Scholar
  37. Zachrisson A, Bornman CH (1984) Application of electric field fusion in plant tissue culture. Physiol Plant 61: 314–320CrossRefGoogle Scholar
  38. Zimmermann U (1982) Electric field-mediated fusion and related phenomena. Biochim Biophys Acta 694: 227–277PubMedGoogle Scholar
  39. Zimmermann U, Scheurich P (1981) High frequency fusion of plant protoplasts by electric fields. Planta 151: 26–32CrossRefGoogle Scholar
  40. Zimmermann U, Buchner KH, Arnold WM (1984) Electrofusion of cells: recent developments and relevance for evolution. In: Allen MJ, Usherwoods PNR (eds) Charge and field effects in biosystems. Abacus Press, Normal, IL, pp 293–318Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1989

Authors and Affiliations

  • G. W. Bates
    • 1
  1. 1.Department of Biological Science and Institute of Molecular BiophysicsFlorida State UniversityTallahasseeUSA

Personalised recommendations