Cell Fusion pp 479-496 | Cite as

Electrofusion and Plant Somatic Hybridization

  • George W. Bates
  • Lawrence J. Nea
  • Clare A. Hasenkampf


The transfer of genetic information between sexually incompatible species creates unique experimental opportunities for geneticists and cell biologists. This is especially true of plant somatic cell fusion because the totipotency of plant cells makes it possible to grow fusion products into mature plants (reviewed by Gleba and Sytnik, 1984; Vasil and Vasil, 1980). Although Carlson et al. (1972) reported recovering the first interspecific plant hybrid through protoplast fusion in 1972, subsequent progress in this field has been slower than originally hoped. This problem is due to several biological and technical barriers. This chapter describes our progress in the application of electrofusion to plant somatic hybridization (see Bates et al., 1983; Bates, 1985; Bates and Hasenkampf, 1985) and suggests solutions to some of the problems in this field.


Somatic Hybrid Protoplast Fusion Fusion Product Mesophyll Protoplast Plant Protoplast 
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  1. Aviv, D., and Galun, E., 1984, The feeder layer technique, in: Cell Culture and Somatic Cell Genetics of Plants, Vol. 1 (I. K. Vasil, ed.), pp. 199–203, Academic Press, New York.Google Scholar
  2. Bates, G. W., 1985, Electrical fusion for optimal formation of protoplast heterokaryons in Nicotiana, Planta 165:217–224.CrossRefGoogle Scholar
  3. Bates, G. W., and Hasenkampf, C. A., 1985, Culture of plant somatic hybrids following electrical fusion, Theor. Appl. Genet.70:277–233.CrossRefGoogle Scholar
  4. Bates, G. W., Gaynor, J. J., and Shekhawat, N. S., 1983, Fusion of plant protoplasts by electric fusion, Plant Physiol 72:1110–1113.PubMedCrossRefGoogle Scholar
  5. Brewbaker, J. L., Upadhya, M. D., Makinen, Y., and MacDonald, T., 1968, Isoenzyme polymorphism in flowering plants. 3. Gel electrophoretic methods and applications, Physiol Plant 121: 930–940.CrossRefGoogle Scholar
  6. Carlson, P. S., Smith, H. H., and Dearing, R. D., 1972, Parasexual interspecific plant hybridization, Proc. Natl Acad. Sci. U.S.A.69:2292–2294.PubMedCrossRefGoogle Scholar
  7. Chapel, M., Montane, M. H., Ranty, B., Teissie, J., and Alibert, G., 1986, Viable somatic hybrids obtained by direct current electrofusion of chemically aggregated plant protoplasts, FEBS Lett.196:79–86.CrossRefGoogle Scholar
  8. Constabel, F., Dudits, D., Gamborg, O. L., and Kao, K. N., 1975a, Nuclear fusion in intergeneric heterokaryons, a note, Can. J. Bot.53:2092–2095.CrossRefGoogle Scholar
  9. Constabel, F., Kirkpatrick, J. W., Kao, K. N., and Kartha, K. K., 1975b, The effect of canavanine on the growth of cells from suspension cultures and on intergeneric heterokaryocytes of canavanine sensitive and tolerant plants, Biochem. Physiol. Pflanz.168:319–325.Google Scholar
  10. Davey, M. R., and Kumar, A., 1983, Higher plant protoplasts—retrospect and prospect, in: Plant Protoplasts, International Review of Cytology, Supplement 16 (K. L. Giles, ed.), pp. 219–299, Academic Press, New York.Google Scholar
  11. Evans, D. A., and Bravo, J. E., 1983, Plant protoplast isolation and culture, in: Plant Protoplasts, International Review of Cytology, Supplement 16 (K. L. Giles, ed.), pp. 33–53, Academic Press, New York.Google Scholar
  12. Flores, H. E., Kaur-Sawhney, R., and Galston, A. W., 1981, Protoplasts as vehicles for plant propagation and improvement, Adv. Cell Culture 1:241–279.Google Scholar
  13. Fluhr, R., Aviv, D., Galun, E., and Edelman, M., 1984, Generation of heteroplastidic Nicotiana cybrids by protoplast fusion: Analysis for plastid recombinant types, Theor. Appl. Genet.67:491–497.CrossRefGoogle Scholar
  14. Fraley, R. T., Rogers, S. G., Horsch, R. B., Sanders, P. R., Flick, J. S., Adams, S. P., Bittner, M. L., Brand, L. A., Fink, C. L., Fry, J. S., Galluppi, G. R., Goldberg, S. B., Hoffmann, N. L., and Woo, S. C., 1983, Expression of bacterial genes in plant cells, Proc. Natl. Acad. Sci. U.S.A.80:4803–4807.PubMedCrossRefGoogle Scholar
  15. Fromm, M., Taylor, L. P., and Walbot, V., 1985, Expression of genes electroporated into mon-ocot and dicot plant cells, Proc. Natl. Acad. Sci. U.S.A.82:5824–5828.PubMedCrossRefGoogle Scholar
  16. Gleba, Y. y, and Sytnik, K. M., 1984, Protoplast Fusion, Springer-Verlag, New York.CrossRefGoogle Scholar
  17. Glimelius, K., Eriksson, T., Grafe, R., and Muller, A. J., 1978, Somatic hybridization of nitrate reductase deficient mutants of Nicotiana tabacum by protoplast fusion, Physiol. Plant.44:273–277.CrossRefGoogle Scholar
  18. Horsch, R. B., and Jones, G. E., 1980, A double filter paper technique for plating cultured plant cells, In Vitro 16: 103–108.CrossRefGoogle Scholar
  19. Horsch, R. B., Fraley, R. T., Rogers, S. G., Sanders, P. R, Lloyd, A., and Hoffmann, N., 1984, Inheritance of functional foreign genes in plants, Science 223:496–498.PubMedCrossRefGoogle Scholar
  20. Horsch, R. B., Fry, J. E., Hoffmann, N. L., Eichholtz, D., Rogers, S. G., and Fraley, R. T., 1985, A simple and general method for transferring genes into plants, Science 227:1129–1231.Google Scholar
  21. Kao, K. N., 1975a, A nuclear staining method for plant protoplasts, in: Plant Tissue Culture Methods (O. L. Gamborg and L. R. Wetter, eds.), pp. 60–62, Prairie Research Laboratory, Saskatoon.Google Scholar
  22. Kao, K. N., 1975b, A method for fusion of plant protoplasts with polyethylene glycol, in: Plant Tissue Culture Methods (O. L. Gamborg and L. R. Wetter, eds.), pp. 22–27, Prairie Research Laboratory, Saskatoon.Google Scholar
  23. Kao, K. N., 1977, Chromosomal behavior in somatic hybrids of soybean—Nicotiana glauca, Mol. Gen. Genet.150:225–230.CrossRefGoogle Scholar
  24. Kao, K. N., 1981, Plant protoplast fusion and somatic hybrids, in: Plant Tissue Culture (H. Han, ed.), pp. 331–339, Proceedings of the Beijing Symposium, Pitman, London.Google Scholar
  25. Kao, K. N., and Michayluk, M. R., 1974, A method for high frequency intergeneric fusion of plant protoplasts, Planta 115: 355–367.CrossRefGoogle Scholar
  26. Kartha, K. K., Gamborg, O. L., Constabel, F., and Kao, K. N., 1974, Fusion of rapeseed and soybean protoplasts and subsequent division of heterokaryocytes, Can. J. Bot.52:2435–2436.CrossRefGoogle Scholar
  27. Kohn, H., Schieder, R., and Schieder, O., 1985, Somatic hybrids in tobacco mediated by electrofusion, Plant Sci.38:121–128.CrossRefGoogle Scholar
  28. Koop, H.-U., 1985, Regeneration of plants after electrofusion of selected pairs of protoplasts, Eur. J. Cell Biol.39:46–49.Google Scholar
  29. Maliga, P., Lazar, G., Joo, F., Nagy, A H., and Menczel, L., 1977, Restoration of morpho-genetic potential in Nicotiana by somatic hybridization, Mol. Gen. Genet.157:291–296.CrossRefGoogle Scholar
  30. Menczel, L., Galiba, G., Nagy, F., and Maliga, P., 1982, Effect of radiation dosage on efficiency of chloroplast transfer by protoplast fusion in Nicotiana, Genetics 100:487–495.Google Scholar
  31. Miller, R. A., Gamborg, O. L., Keller, W. A., and Kao, K.N., 1971, Fusion and division of nuclei in multinculeated soybean protoplasts, Can. J. Genet. Cytol.13:347–353.Google Scholar
  32. Morikawa, H., Sugino, K., Hayashi, Y., Takeda, J., Senda, M., Hirai, A., and Yamada, Y., 1986, Interspecific plant hybridization by electrofusion in Nicotiana, Biotechnology 4: 57–60.CrossRefGoogle Scholar
  33. Nagy, J. I., and Maliga, P., 1976, Callus induction and plant regeneration from protoplasts of Nicotiana sylvestris, Z. Pflanzenphysiol.78:453–455.Google Scholar
  34. Negrutiu, I., DeBrouwer, D., Watts, J. W., Sidorov, V. I., Dirks, R., and Jacobs, M., 1986, Fusion of plant protoplasts: a study using auxotrophic mutants of Nicotiana plumbaginifolia Viviani, Theor. Appl. Genet.72:279–286.CrossRefGoogle Scholar
  35. Pohl, H. A., 1978, Dielectrophoresis, Cambridge University Press, Cambridge.Google Scholar
  36. Potrykus, I., Paszkowsi, J., Saul, M. W., Petruska, J., and Shillito, R. D., 1985a, Molecular and general genetics of a hybrid foreign gene introduced into tobacco by direct gene transfer, Mol. Gen. Genet.199:169–177.PubMedCrossRefGoogle Scholar
  37. Potrykus, I., Saul, M. W., Petruska, J., Paszkowski, J., and Shillito, R. D., 1986, Direct gene transfer to cells of a graminaceous monocot, Mol. Gen. Genet.199:183–188.CrossRefGoogle Scholar
  38. Power, J. B., Berry, S. F., Frearson, E. M., and Cocking, E. C., 1977, Selection procedures for the production of inter-species somatic hybrids of Petunia hybrida and Petunia parodii, Plant Sci. Lett.7:51–55.Google Scholar
  39. Puite, K. J., van Wikselaar, P., and Verhoeven, H., 1985, Electrofusion, a simple and reproducible technique in somatic hybridization of Nicotiana plumbaginifolia mutants, Plant Cell Rep.4:274–276.CrossRefGoogle Scholar
  40. Scheurich, P., and Zimmermann, U., 1981, Giant human erythrocytes by electric-field-induced cell-to-cell fusion, Naturwissenschaften 68: 45–47.PubMedCrossRefGoogle Scholar
  41. Shillito, R. D., Paszkowski, J., and Potrykus, I., 1983, Agarose plating and a bead type culture technique enable and stimulate development of protoplast-derived colonies in a number of plant species, Plant Cell Rep.2:244–247.CrossRefGoogle Scholar
  42. Sidorov, V. A., Menczel, L., Nagy, F., and Maliga, P., 1981, Chloroplast transfer in Nicotiana based on metabolic complementation between irradiated and iodoacetate treated protoplasts, Planta 152: 341–345.CrossRefGoogle Scholar
  43. Tempelaar, M. J., and Jones, M. G. K., 1985a, Directed electrofusion between protoplasts with different responses in a mass fusion system, Plant Cell Rep.4:92–95.CrossRefGoogle Scholar
  44. Tempelaar, M. J., and Jones, M. G. K., 1985b, Fusion characteristics of plant protoplasts in electric fields, Planta 165: 205–216.CrossRefGoogle Scholar
  45. Vasil, I. K., and Vasil, V., 1980, Isolation and culture of protoplasts, Int. Rev. Cytol. (Suppl.) 11A: 1–20.CrossRefGoogle Scholar
  46. Vienken, J., and Zimmermann, U., 1982, Electric field-induced fusion: Electro-hydraulic procedure for production of heterokaryon cells in high yield, FEBS Lett.137:11–13.PubMedCrossRefGoogle Scholar
  47. Watts, J. W., and King, J. M., 1984, A simple method for large-scale electrofusion and culture of plant protoplasts, Biosci. Rep.4:335–342.PubMedCrossRefGoogle Scholar
  48. Zimmermann, U., 1982, Electric field-mediated fusion and related electrical phenomena, Biochim. Biophys. Acta 694:227–277.PubMedCrossRefGoogle Scholar
  49. Zimmermann, U., and Scheurich, P., 1981, High frequency fusion of plant protoplasts by electric fields, Planta 151: 26–32.CrossRefGoogle Scholar
  50. Zimmermann, U., Buchner, K. H., and Arnold, W. M., 1984, Electrofusion of cells: recent developments and relevance for evolution, in: Charge and Field Effects in Biosystems (M. J. Allen and P. N. R. Usherwood, eds.), pp. 293–318, Abacus Press, Normal, Ill.Google Scholar

Copyright information

© Springer Science+Business Media New York 1987

Authors and Affiliations

  • George W. Bates
    • 1
  • Lawrence J. Nea
    • 1
  • Clare A. Hasenkampf
    • 1
  1. 1.Department of Biological Science and Institute of Molecular BiophysicsFlorida State UniversityTallahasseeUSA

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