Fusion of Somatic Cells

  • T. Nagata
Part of the Encyclopedia of Plant Physiology book series (PLANT, volume 17)


At the onset it should be noticed that the contents of this chapter are confined to the fusion of protoplasts; normal somatic plant cells with thick cell walls cannot fuse with each other and only naked protoplasts can do so. Nowadays fusion of protoplasts has attracted attention as the first step towards somatic hybridization, but the phenomenon is also interesting from the view of plant physiology. The first report of the fusion of protoplasts was by KüSter (1909, 1910). He observed the fusion of protoplasts which were isolated from the epidermis of tulip bulbs according to the mechanical method of KLercker (1892). Later MIchel (1937) observed the fusion between protoplasts from different species and tissues. These protoplasts were also isolated mechanically from the tissues plasmolyzed previously with 0.5 M KNO3; however, when sucrose was employed as a plasmolyticum, fusion was not observed. In these pioneering studies the work of MIchel is especially important, because fusion could be induced not only between the protoplasts from the same tissue, but also between protoplasts from different tissues and different plants. However, these interesting observations went unnoticed until the report of POwer et al. (1970). This neglect may be due to the fact that at the time of MIchel the technique of plant cell culture had not yet been established and the yield of protoplasts from tissues was extremely restricted because of the mechanical method utilized. The new possibility for the development of protoplast research started from the report of COcking (1960), who isolated protoplasts from root tissues of tomato by a self–made crude cellulase preparation from a fungus, Myrothecium verrucaria.


Polyethylene Glycol Membrane Fluidity Somatic Hybridization Cell Fusion Membrane Fusion 
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. Bajaj YPS (1977) Protoplast isolation, culture and somatic hybridization. In: Reinert J, Bajaj YPS (eds) Applied and fundamental aspects of plant cell, tissue, and organ culture. Springer, Berlin Heidelberg New York, pp 467–577Google Scholar
  2. Bangham AD, Flemans R, Heard DH, Seaman GVF (1958) An apparatus for microelectrophoresis of small particles. Nature 182: 642–644PubMedCrossRefGoogle Scholar
  3. Bawa SB, Torrey JG (1971) “Budding” and nuclear division in cultured protoplasts of corn, convolvulus, and onion. Bot Gaz 132:240–245Google Scholar
  4. Blow AMJ, Botham GM, Fisher D, Goodall AH, Tilcock CPS, Lucy JA (1978) Water and calcium in cell fusion induced by poly(ethylene glycol). FEBS Lett 94: 305–310PubMedCrossRefGoogle Scholar
  5. Borochov A, Borochov H (1979) Increase in membrane fluidity in liposomes and plant protoplasts upon osmotic swelling. Biochim Biophys Acta 550: 546–549PubMedCrossRefGoogle Scholar
  6. Boss WF, Mott RL (1980) Effect of divalent cations and polyethylene glycol on the membrane fluidity of protoplasts. Plant Physiol 66: 835–837PubMedCrossRefGoogle Scholar
  7. Burgess J, Linstead PJ (1976) Ultrastructural studies of the binding of concanavalin A to the plasmalemma of higher plant protoplasts. Planta 130: 73–79CrossRefGoogle Scholar
  8. Carlson PS, Smith HH, Dearing RD (1972) Parasexual interspecific plant hybridization. Proc Natl Acad Sci USA 69: 2292–2294PubMedCrossRefGoogle Scholar
  9. Cocking EC (1960) A method for the isolation of plant protoplasts and vacuoles. Nature 187: 962–963CrossRefGoogle Scholar
  10. Cocking EC (1972) Plant cell protoplasts–isolation and development. Annu Rev Plant Physiol 23: 29–50CrossRefGoogle Scholar
  11. Cocking EC (1977) Selection and somatic hybridization. In: Thorpe TA (ed) Frontiers of plant tissue culture 1978. The Bookstore, Univ Calgary, Calgary pp 151–158Google Scholar
  12. Constabel F (1975) Protoplast isolation. In: Gamborg OL, Wetter LR (eds) Plant tissue culture methods. National Research Council of Canada, Ottawa, pp 11–21Google Scholar
  13. Curtis ASG (1960) Cell contacts: Some physical considerations. Am Nat 94: 37–56CrossRefGoogle Scholar
  14. Dudits D, Rasko I, Jadlaczky GY, Lima–de–Faria A (1976) Fusion of human cells with carrot protoplasts induced by polyethylene glycol. Hereditas 82: 121–124Google Scholar
  15. Eriksson T, Glimelius K, Wallin A (1978) Protoplast isolation, cultivation and development. In: Thorpe TA (ed) Frontiers of plant tissue culture 1978. The Bookstore, Univ Calgary, Calgary, pp 131–139Google Scholar
  16. Eyler EH (1962) The contribution of sialic acid to the surface charge of the erythrocyte. J Biol Chem 237: 1992–2000Google Scholar
  17. Fowke LC, Marchant HJ, Gresshoff PM (1981) Fusion of protoplasts from carrot cell cultures and the green alga Stigeoclonium. Can J Bot 59: 1021–1025CrossRefGoogle Scholar
  18. Galun E (1981) Plant protoplasts as physiological tools. Annu Rev Plant Physiol 32: 237–266CrossRefGoogle Scholar
  19. Gieba YY, Hoffmann F (1980) “Arabidobrassica”: A novel plant obtained by protoplast fusion. Planta 149:112–117 Glimelius K, Wallin A, Eriksson T (1974) Agglutination effects of concanavalin A onGoogle Scholar
  20. isolated protoplasts of Daucus carota. Physiol Plant 31:225–230Google Scholar
  21. Glimelius K, Wallin A, Eriksson T (1978) Concanavalin A improves the polyethylene glycol method for fusing plant protoplasts. Physiol Plant 44: 92–96CrossRefGoogle Scholar
  22. Grout BWW, Willison JHM, Cocking EC (1973) Interactions at the surface of plant cell protoplasts; An electrophoretic and freeze–etch study. J Bioenerget 4: 586–611Google Scholar
  23. Hasezawa S, Nagata T, Syono K (1981) Transformation of Vinca protoplasts mediated by Agrobacterium spheroplasts. Mol Gen Genet 182: 206–210CrossRefGoogle Scholar
  24. Haydon DA, Seaman GVF (1962) An estimation of the surface ionogenic groups of the human erythrocyte and of Escherichia coli. Proc R Soc Lond Ser B 156: 533–549CrossRefGoogle Scholar
  25. Hopwood HA (1981) Genetic studies with bacterial protoplasts. Annu Rev Microbiol 35: 237–272PubMedCrossRefGoogle Scholar
  26. Inbar M, Sachs L (1969) Interaction of the carbohydrate–binding protein concanavalin A with normal and transformed cells. Proc Natl Acad Sei USA 63: 1418–1425CrossRefGoogle Scholar
  27. Itoh M (1973) Studies on the behavior of meiotic protoplasts. II. Induction of a high fusion frequency in protoplasts from liliaceous plants. Plant Cell Physiol 14: 865–872Google Scholar
  28. Jones MN (1975) Biological interfaces. Elsevier, North/Holland, Amsterdam Oxford New YorkGoogle Scholar
  29. Jones CW, Mastrangelo IA, Smith HH, Liu HZ (1976) Interkingdom fusion between human (HeLa) cells and tobacco hybrid ( GGLL) protoplasts. Science 193: 401–403Google Scholar
  30. Kao KN, Michayluk MR (1974) A method for high–frequency intergeneric fusion of plant protoplasts. Planta 115: 355–367CrossRefGoogle Scholar
  31. Kao KN, Constabel F, Michayluk MR, Gamborg OL (1974) Plant protoplasts fusion and growth of intergeneric hybrid cells. Planta 120: 215–227CrossRefGoogle Scholar
  32. Keller WA, Melchers G (1973) The effect of high pH and calcium on tobacco leaf protoplast fusion. Z Naturforsch 28 c: 737–741Google Scholar
  33. Klercker J, (1892) Eine Methode zur Isolierung lebender Protoplasten. Oefversigt Kongl Vetens–Akad Foerhandlingar 49: 463–474Google Scholar
  34. Knutton S, Pasternak CA (1979) The mechanism of cell–cell fusion. Trends Biochem Sei 4: 220–223CrossRefGoogle Scholar
  35. Küster E (1909) Über die Verschmelzung nackter Pro toplasten. Ber Dtsch Bot Ges 27: 589–598Google Scholar
  36. Küster E (1910) Eine Methode zur Gewinnung abnorm großer Protoplasten. Arch Entw Mechan 30: 351–355Google Scholar
  37. Larkin PJ (1978) Plant protoplast agglutination by lectins. Plant Physiol 61: 626–629PubMedCrossRefGoogle Scholar
  38. Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18: 100–127CrossRefGoogle Scholar
  39. Maggio B, Ahkong QF, Lucy JA (1976) Polyethylene glycol), surface potential and cell fusion. Biochem J 158: 647–650PubMedGoogle Scholar
  40. Melchers G (1977) Plant hybrids by fusion of protoplasts. In: Beers Jr RF, Bassett EG (eds) Recombinant molecules: Impact on science and society. Raven Press, New York, pp 209–227Google Scholar
  41. 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: 277–294Google Scholar
  42. Melchers G, Sacristan MD, Holder AA (1978) Somatic hybrid plants of potato and tomato regenerated from fused protoplasts. Carlsberg Res Comm 43: 203–218CrossRefGoogle Scholar
  43. Michel W (1937) Über die experimentelle Fusion pflanzlicher Pro toplasten. Arch Exp Zellforsch 20: 230–252Google Scholar
  44. Nagata T (1978) A novel cell–fusion method of protoplasts by polyvinyl alcohol. Naturwissenschaften 65: 263–264CrossRefGoogle Scholar
  45. Nagata T (1983) Interaction of liposomes and protoplasts as a model system of protoplast fusion. In: Beers Jr RF, Bassett EG (eds) Cell fusion: Gene transfer and transformation. Raven Press, New York, pp 217–226Google Scholar
  46. Nagata T, Ishii S (1979) A rapid method for isolation of mesophyll protoplasts. Can J Bot 57: 1820–1823CrossRefGoogle Scholar
  47. Nagata T, Melchers G (1978) Surface charges of protoplasts and their significance in cell–cell interaction. Planta 142: 235–238CrossRefGoogle Scholar
  48. Nagata T, Takebe I (1970) Cell wall regeneration and cell division in isolated tobacco mesophyll protoplasts. Planta 92: 12–20CrossRefGoogle Scholar
  49. Nagata T, Takebe I (1971) Plating of isolated tobacco mesophyll protoplasts on agar medium. Planta 99: 12–20CrossRefGoogle Scholar
  50. Nagata T, Eibl H, Melchers G (1979) Fusion of plant protoplasts by a positively charged synthetic phospholipid. Z Naturforsch 34c: 460–462Google Scholar
  51. Nagata T, Okada K, Takebe I, Matsui C (1981) Delivery of tobacco mosaic virus RNA into plant protoplasts mediated by reverse–phase evaporation vesicles (liposomes). Mol Gen Genet 184: 161–165Google Scholar
  52. Ohshima H (1975) Diffuse double layer interaction between two spherical particles with constant surface charge density in an electrolyte solution. Colloid Polymer Sei 263: 158–163CrossRefGoogle Scholar
  53. Peberdy JF (1979) Fungal protoplasts: Isolation, reversion, and fusion. Annu Rev Microbiol 33: 21–39PubMedCrossRefGoogle Scholar
  54. Pontecorvo G, Riddle PN, Hales A (1977) Time and mode of fusion of human fibroblasts treated with polyethylene glycol ( PEG ). Nature 265: 257–258Google Scholar
  55. Poste G, Allison AC (1973) Membrane fusion. Biochim Biophys Acta 300: 421–465PubMedGoogle Scholar
  56. Potrykus I (1971) Intra and interspecific fusion of protoplasts from petals of Torenia beillonii and Torenia fournieri. Nat New Biol 231: 57–58PubMedCrossRefGoogle Scholar
  57. Power JB, Cummins SE, Cocking EC (1970) Fusion of isolated protoplasts. Nature 225: 1016–1018PubMedCrossRefGoogle Scholar
  58. Power JB, Berry SF, Chapman JV, Cocking EC (1980) Somatic hybridization of sexually incompatible petunias: Petunia parodii, Petunia parviflora. Theor Appl Genet 54: 1–4Google Scholar
  59. Robenek H, Peveling E (1978) Beobachtungen am Plasmalemma während der Fusion isolierter Protoplasten von Skimmia japonica Thunb. Ber Dtsch Bot Ges 91: 351–359Google Scholar
  60. Robinson JM, Roos DS, Davidson RL, Karnovsky MJ (1979) Membrane alterations and other morphological features associated with polyethylene glycol–induced cell fusion. J Cell Sei 40: 63–75Google Scholar
  61. Rutishauser U, Sachs L (1975) Cell–to–cell binding induced by different lectins. J Cell Biol 65: 247–257PubMedCrossRefGoogle Scholar
  62. Schieder O (1978) Somatic hybrids of Datura innoxia Mill. +Datura discolor Bernh. and of Datura innoxia Mill. Datura stramonium L. var tabula L. Mol Gen Genet 162: 113–119CrossRefGoogle Scholar
  63. Schieder O, Vasil IK (1980) Protoplast fusion and somatic hybridization. In: Vasil I (ed) Recent advances in plant cell and tissue culture. Int Rev Cytol Suppl IIB, Academic Press, London New York, pp 21–46Google Scholar
  64. Smith BA, Ware BR, Weiner RS (1976) Electrophoretic distributions of human peripheral blood mononuclear white cells from normal subjects and from patients with acute lymphocytic leukemia. Proc Natl Acad Sci USA 73: 2388–2391PubMedCrossRefGoogle Scholar
  65. Takebe I, Labib G, Melchers G (1971) Regeneration of whole plants from isolated mesophyll protoplasts of tobacco. Naturwissenschaften 58: 318–320CrossRefGoogle Scholar
  66. Takebe I, Otsuki Y, Aoki S (1968) Isolation of tobacco mesophyll cells in intact and active state. Plant Cell Physiol 9: 115–124Google Scholar
  67. Trauble H, Eibl H (1974) Electrostatic effects on lipid phase transitions: Membrane structure and ionic environment. Proc Natl Acad Sci USA 71: 214–219Google Scholar
  68. Wallin A, Glimelius K, Eriksson T (1974) The induction and fusion of Daucus carota protoplasts by polyethylene glycol. Z Pflanzenphysiol 74: 64–80Google Scholar
  69. Ward M, Davey MR, Mathias RJ, Cocking EC, Clothier RH, Balls M, Lucy JA (1979) Effects of pH, Ca2 +, temperature and protease pretreatment on interkingdom fusion. Somat Cell Genet 5: 529–536PubMedCrossRefGoogle Scholar
  70. Wilkins DJ, Ottewill RH, Bangham AD (1962) On the flocculation of sheep leucocytes: I. Electrophoretic studies. J Theoret Biol 2: 165–175Google Scholar
  71. Wilschut J, Diizgiines N, Fraley R, Papahadjopoulos D (1980) Studies on the mechanism of membrane fusion: Kinetics of calcium ion–induced fusion of phosphatidylserine vesicles followed by a new assay for mixing of aqueous vesicle contents. Biochemistry 19: 6011–6021Google Scholar
  72. Withers LA, Cocking EC (1972) Fine–structural studies on spontaneous and induced fusion of higher plant protoplasts. J Cell Sci 11: 59–75PubMedGoogle Scholar
  73. Zimmermann U, Scheurich P (1981) High frequency fusion of plant protoplasts by electric fields. Planta 151: 26–32CrossRefGoogle Scholar
  74. Zimmermann U, Scheurich P, Pilwat G, Benz R (1981) Cells with manipulated functions: New perspectives for cell biology, medicine, and technology. Angew Chem 20: 325–344Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • T. Nagata

There are no affiliations available

Personalised recommendations