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Electrofusion: The Technique and Its Application to Somatic Hybridization

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Plant Protoplasts and Genetic Engineering II

Part of the book series: Biotechnology in Agriculture and Forestry ((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.

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References

  • Bates GW (1985) Electrical fusion for optimal formation of protoplast heterokaryons in Nicotiana. Planta 165: 217–224

    Article  Google Scholar 

  • Bates GW, Gaynor JJ, Shekhawat NS (1983) Fusion of plant protoplasts by electric fields. Plant Physiol 72: 1110–1113

    Article  PubMed  CAS  Google Scholar 

  • Bates GW, Hasenkampf CA (1985) Culture of plant somatic hybrids following electrical fusion. Theor Appl Genet 70: 227–233

    Article  Google Scholar 

  • Bates GW, Hasenkampf CA, Contolini CS, Piastuch WC (1987a) Limited gene transfer by somatic cell fusion in Nicotiana. Theor Appl Genet 74: 718–726

    Article  Google Scholar 

  • Bates GW, Nea LJ, Hasenkampf CA ( 1987 b) Electrofusion and plant somatic hybridization. In: Sowers AE (ed) Cell fusion. Plenum, New York, pp 479–496

    Google Scholar 

  • Bates GW, Saunders JA, Sowers AE (1987c) Electrofusion, principles and applications. In: Sowers AE (ed) Cell fusion. Plenum, New York, pp 367–395

    Google Scholar 

  • Chapel M, Teissie J, Alibert G (1984) Electrofusion of spermine-treated plant protoplasts. FEBS Lett 173: 331–336

    Article  CAS  Google Scholar 

  • 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: 79–86

    Article  CAS  Google Scholar 

  • Constabel F, Dudits D, Gamborg OL, Kao KN (1975) Nuclear fusion in intergeneric heterokaryons. A note. Can J Bot 53: 2092–2095

    Article  Google Scholar 

  • 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 211–222

    Google Scholar 

  • Fromm ME, Taylor LP, Walbot V (1986) Stable transformation of maize after gene transfer by electroporation. Nature 319: 791–793

    Article  PubMed  CAS  Google Scholar 

  • 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 385

    Google Scholar 

  • 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 22–27

    Google Scholar 

  • Kinosita K, Tsong TY (1977) Formation and resealing of pores of controlled sizes in human erythrocyte membrane. Nature 268: 438–441

    Article  PubMed  Google Scholar 

  • Kinosita K, Tsong TY (1979) Voltage-induced conductance in human erythrocyte membranes. Biochim Biophys Acta 554: 479–497

    Article  PubMed  CAS  Google Scholar 

  • Kohn H, Schieder HR, Schieder O (1985) Somatic hybrids in tobacco mediated by electrofusion. Plant Sci 38: 121–128

    Article  Google Scholar 

  • Koop H-U, Schweiger H-G (1985) Regeneration of plants after electrofusion of selected pairs of protoplasts. Eur J Cell Biol 39: 46–49

    Google Scholar 

  • 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: 89–94

    Article  CAS  Google Scholar 

  • Morikawa H, Sugino K, Hayashi Y, Takeda J, Senda M, Hirai A, Yamada Y (1986) Interspecific plant hybridization by electrofusion in Nicotiana. Biotechnology 4: 57–60

    Article  Google Scholar 

  • 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: 419–422

    Article  Google Scholar 

  • Nea LJ, Bates GW (1987) Protoplast electrofusion: factors affecting fusion efficiency and cell viability. Plant Cell Rep 6: 337–340

    Article  CAS  Google Scholar 

  • Nea LJ, Bates GW, Gilmer PJ (1987) Facilitation of electrofusion of plant protoplasts by membrane active agents. Biochim Biophys Acta 897: 293–301

    Article  CAS  Google Scholar 

  • 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: 279–286

    Article  Google Scholar 

  • Pohl HA (1978) Dielectrophoresis. Cambridge Univ Press, London

    Google Scholar 

  • 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: 274–276

    Article  Google Scholar 

  • Puite KJ, Roest S, Pijnacker LP (1986) Somatic hybrid potato plants after electrofusion of diploid Solanum tuberosum and Solanum phureja. Plant Cell Rep 5: 262–265

    Article  CAS  Google Scholar 

  • Riggs CD, Bates GW (1986) Stable transformation of tobacco by electroporation: evidence for plasmid concatenation. Proc Natl Acad Sci USA 83: 5602–5606

    Article  PubMed  CAS  Google Scholar 

  • Ruzin SE, McCarthy SC (1986) The effect of chemical facilitators on the frequency of electrofusion of tobacco mesophyll protoplast. Plant Cell Rep 5: 342–345

    Article  CAS  Google Scholar 

  • Scheurich P, Zimmermann U (1981) Electrically stimulated fusion of different plant cell protoplasts. Plant Physiol 67: 849–853

    Article  PubMed  CAS  Google Scholar 

  • Senda M, Takeda J, Abe S, Nakamura T (1979) Induction of cell fusion of plant protoplasts by electrical stimulation. Plant Cell Physiol 20: 1441–1443

    CAS  Google Scholar 

  • 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: 51–56

    Google Scholar 

  • Tempelaar MJ, Jones MGK (1985 a) Fusion characteristics of plant protoplasts in electric fields. Planta 165: 205–216

    Article  Google Scholar 

  • Tempelaar MJ, Jones MGK (1985 b) Directed electrofusion between protoplasts with different responses in a mass fusion system. Plant Cell Rep 4: 92–95

    Article  Google Scholar 

  • 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: 99–105

    Article  CAS  Google Scholar 

  • 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: 64–70

    Article  Google Scholar 

  • Watts JW, King JM (1984) A simple method for large-scale electrofusion and culture of plant protoplasts. Biosci Rep 4: 335–342

    Article  PubMed  CAS  Google Scholar 

  • Zachrisson A, Bornman CH (1984) Application of electric field fusion in plant tissue culture. Physiol Plant 61: 314–320

    Article  Google Scholar 

  • Zimmermann U (1982) Electric field-mediated fusion and related phenomena. Biochim Biophys Acta 694: 227–277

    PubMed  CAS  Google Scholar 

  • Zimmermann U, Scheurich P (1981) High frequency fusion of plant protoplasts by electric fields. Planta 151: 26–32

    Article  Google Scholar 

  • 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 293–318

    Google Scholar 

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© 1989 Springer-Verlag Berlin Heidelberg

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Bates, G.W. (1989). Electrofusion: The Technique and Its Application to Somatic Hybridization. In: Bajaj, Y.P.S. (eds) Plant Protoplasts and Genetic Engineering II. Biotechnology in Agriculture and Forestry, vol 9. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-74454-9_15

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  • DOI: https://doi.org/10.1007/978-3-642-74454-9_15

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-74456-3

  • Online ISBN: 978-3-642-74454-9

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