Somatic Hybridization of Plants and its Use in Agriculture

  • John D. Hamill
  • Edward C. Cocking
Chapter
Part of the NATO ASI Series book series (volume 18)

Abstract

Since Carlson et al. (1972) first reported the successful recovery of somatic hybrid plants, very many protoplast fusion experiments have been done. These have ranged from experiments aimed at answering fundamental questions on the interaction of different plant genomes following fusion to practical questions regarding the improvement of crop species by fusion with wild species possessing desirable agronomic traits. The agricultural applications of protoplast fusion are now beginning to be realised and although the recent successes in plant transformation offer many new opportunities in plant genetic engineering for crop improvement, it is clear that protoplast fusion does have an important role to play in practical plant biotechnology.

In this article we discuss the methodologies needed to recover somatic hybrids, their key features and some examples of somatic hybrids which are proving useful in agriculture. It should be noted that there exists a huge literature on protoplast fusion and only a limited number of examples are quoted in each section.

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References

  1. Abduallah R, Cocking EC, Thompson JA (1986) Efficient plant regeneration from rice protoplasts through somatic embryogenesis. Biotechnology 4: 1087–1090CrossRefGoogle Scholar
  2. Afonso CL, Harkins KR, Thomas-Compton MA, Krejci AE, Gabraith DW (1985) Selection of somatic hybrid plants in Nicotiana through fluorescence activated cell sorting. Biotechnol 3: 811–816CrossRefGoogle Scholar
  3. Aird ELH, Hamill JD, Rhodes MJC (1987) Cytogenetic analysis of a number of plant species transformed by Agrobacterium rhizogenes. Theor Appl Genet (submitted)Google Scholar
  4. Austin S, Baer MA, Helgeson JP (1985) Transfer of resistance to potato leaf roll virus from Solanum brevidens into Solanum tuberosum by protoplast fusion. Plant Sci 39: 75–82CrossRefGoogle Scholar
  5. Belivanis T, Dore C (1986) Interspecific hybridization of Phaseolus vulgaris L. and Phaseolus argustissimus A. Gray using in vitro embryo culture. Plant Cell Rep 5: 329–331CrossRefGoogle Scholar
  6. Belliard G, Vedel F, Pelletier G (1979) Mitochondrial recombination in cytoplasmic hybrids of Nicotiana tabacum by protoplast fusion. Nature 281: 401–403CrossRefGoogle Scholar
  7. Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nuc Acid Res 12: 8711–8721CrossRefGoogle Scholar
  8. Bonnett HT, Glimeluis K (1983) Somatic hybridization in Nicotiana; behaviour of organelles after fusion of protoplasts from male fertile + male sterile cultivars. Theor App Genet 65: 213–217CrossRefGoogle Scholar
  9. Brunold C, Kruger-Lebus S, Saul M, Wegmuller S, Potrykus (1987) Combination of kanamycin resistance and nitrate reductase deficiency as selectable markers in one nuclear genome provides a universal somatic hybridizer in plants. Mol Gen Genet 208: 469–473CrossRefGoogle Scholar
  10. Carlson P, Smith HH, Dearing RD (1972) Parasexual interspecific plant hybridization. Proc Natl Acad Sci USA, 69: 2292–2294PubMedCrossRefGoogle Scholar
  11. Chien YC, Kao KN, Wetter LR (1982) Chromosomal and isozyme studies of Nicotiana tabacumGlycine max hybrid cell lines. Theor Appl Genet 62: 301–304Google Scholar
  12. Chen K, Wildman SG, Smith HH (1977) Chloroplasts DNA distribution in parasexual hybrids as shown by polypeptide composition of fraction 1 protein. 74: 5109–5112Google Scholar
  13. Clark EM, Izhar S, Hanson MR (1985) Independent segregation of the plastid genome and cytoplasmic male sterility in Petunia Somatic hybrids. 199: 440–445Google Scholar
  14. D’Amato F (1985) Cytogenetics of plant cell and tissue cultures and their regenerants. CRC Crit Rev Plant Sci 3: 73–112CrossRefGoogle Scholar
  15. Douglas GC, Keller WA, Setterfield G (1981) Somatic hybridization between Nicotiana rustica and N. tabacum. Can J Bot 59: 220–227CrossRefGoogle Scholar
  16. Doyle GG (1986) Aneuploidy and inbreeding depression in random mating and self-fertilizing autotetraploid populations. Theor Appl Genet 72: 799–806CrossRefGoogle Scholar
  17. Dudits D, Kao KN, Constabel F, Gamborg OL (1976) Fusion of carrot and barley protoplasts and division of heterokaryocytes. Can J Genet Cytol 18: 263–269Google Scholar
  18. Ehlenfeldt MK, Helgeson JP (1987) Fertility of somatic hybrids from protoplast fusions of Solanum brevidens and S. tuberosum. Theor Appl Genet 73: 395–402CrossRefGoogle Scholar
  19. Fraley RT, Rogers SG, Horsch RB, Sanders P, Flick R, Adams JS, Bittner ML, Brand LA, Fink CL, Fry JS, Galluppi GR, Goldberg SB, Hoffmann NL, Woo SC (1983) Expression of bacterial genes in plant cells. Proc Natl Acad Sci USA, 80: 4803–4807PubMedCrossRefGoogle Scholar
  20. Galbraith DW, Mauch TJ (1980) Identification of fusion of plant protoplasts II: Conditions for the reproducible fluorescence labelling of protoplasts derived from mesophyll tissue. Z Pflanzenphysiol 98: 129–140Google Scholar
  21. Galun E, Arzee-Gonen P, Fluhr R, Edelman M, Aviv D (1982) Cytoplasmic hybridization in Nicotiana. Mol Gen Genet 186: 50–56PubMedCrossRefGoogle Scholar
  22. Gilmour DM, Davey MR, Cocking EC, Pental D (1987) Culture of low numbers of forage legume protoplasts in membrane chambers. J Plant Physiol 126: 457–465Google Scholar
  23. Gleba YY, Hoffmann F (1980) “Arabidobrassica” — A novel plant obtained by protoplast fusion. Planta 149: 112–117CrossRefGoogle Scholar
  24. Gleba YY, Kanevsky IF, Skarzhynskaya MV, Komarnitsky IK, Cherep NN (1986) Hybrids between tobacco crown gall cells and normal somatic cells of Atropa belladonna. Plant Cell Rep 5: 394–397CrossRefGoogle Scholar
  25. Glimelius K, Bonnett HT (1986) Nicotiana cybrids with Petunia chloroplasts. Theor Appl Genet 72: 794–798CrossRefGoogle Scholar
  26. Glimelius K, Eriksson T, Grafe R, Muller AJ (1978) Somatic hybridization of nitrate reductase deficient mutants of Nicotiana tabacum by protoplast fusion. Physiol Plant 44: 273–277CrossRefGoogle Scholar
  27. Hamill JD, Pental D, Cocking EC, Müller AJ (1983) Production of a nitrate reductase deficient streptomycin resistant mutant of Nicotiana tabacum for somatic hybridization studies. Heredity 50: 197–200CrossRefGoogle Scholar
  28. Hamill JD, Pental D, Cocking EC (1984) The combination of a nitrate reductase deficient nuclear genome with a streptomycin resistant chloroplast genome, in Nicotiana tabacunty by protoplast fusion. J Plant Physiol 115: 253–261Google Scholar
  29. Hamill JD, Patnaik G, Pental D, Cocking EC (1984) The culture of manually isolated heterokaryons of Nicotiana tabacum and Nicotiana rustica. Proc Ind Acad Sci (Plant Sci) 93: 317–327Google Scholar
  30. Hamill JD, Pental D, Cocking EC (1985) Analysis of fertility in somatic hybrids of Nicotiana rustica and N. tabacum and projeny over two sexual generations. Theor Appl Genet 71: 486–490CrossRefGoogle Scholar
  31. Hamill JD, Parr AJ, Robins RJ, Rhodes MJC (1986) Secondary product formation by cultures of Beta vulgaris and Nicotiana rustica transformed with Agrobacterium rhizogenes. Plant Cell Reports 5: 111–114CrossRefGoogle Scholar
  32. Hamill JD, King JM, Watts JW (1987) Somatic hybridization between Nicotiana tabacum and Nicotiana plumbaginifolia by electrofusion of mesophyll protoplasts. J Plant Physiol 129: 111–118Google Scholar
  33. Handley LW, Nickels RL, Cameron MW, Moore PP, Sink KC (1986) Somatic hybrid plants between Lycopersicon esculentum and Solanum lycopersicoides. Theor Appl Genet 71: 691–697CrossRefGoogle Scholar
  34. Hein T, Przewozny T, Schieder O (1983) Culture and selection of somatic hybrids using an auxotrophic cell line. Theor Appl Genet 64: 119–122CrossRefGoogle Scholar
  35. Helgeson JP, Hunt GJ, Haberlach GT, Austin S (1986) Somatic hybrids between Solanum brevidens and Solanum tuberosum: Expression of a late blight resistance gene and potato leaf roll resistance. Plant Cell Rep 5: 212–214CrossRefGoogle Scholar
  36. Iwai S, Kishi C, Nakata K, Kawashima N (1986) Production of Nicotiana tabacum x Nicotiana acuminata hybrids by ovule culture. Plant Cell Rep 5: 403–404CrossRefGoogle Scholar
  37. Johnston SA, Hanneman RE (1982) Manipulations of endosperm balance number overcome crossing barriers between diploid Solanum species. Science 217: 446–448PubMedCrossRefGoogle Scholar
  38. Kanchanapoom K, Boss WF (1986) The effect of fluorescent labeling on calcium induced fusion of fusogenic carrot protoplasts. Plant Cell Rep 5: 252–255CrossRefGoogle Scholar
  39. Kao KN (1977) Chromosomal behaviour in somatic hybrids of soybean — Nicotiana glauca. Mol Gen Genet 150: 225–230CrossRefGoogle Scholar
  40. Kao KN, Michayluk MR (1974) A method for high frequency intergeneric fusion of plant protoplasts. Planta 115: 355–367CrossRefGoogle Scholar
  41. Kao KN, Saleem M (1986) Improved fusion of mesophyll and cotyledon protoplasts with PEG and high pH Ca solutions. J Plant Physiol 122: 217–225Google Scholar
  42. Keller WA, Melchers G (1973) The effect of high pH and calcium on tobacco leaf protoplast fusion. Z Naturforsch 28C: 737–741Google Scholar
  43. Keller WA, Setterfield G, Douglas GC, Gleddie S, Nakamura C (1982) Production, characterization and utilization of somatic hybrids of higher plants, In Tomes DT, Ellis BE, Harney PM, Kasha KJ and Peterson RL (eds) Application of Plant Cell and Tissue culture to Agriculture and Industry, Guelph 81–114Google Scholar
  44. Kemble RJ, Barsby TL, Wong RSC, Shepard JF (1986) Mitochondrial DNA rearrangements in somatic hybrids of Solanum tuberosum and Solanum brevidens. Theor Appl Genet 72: 787–793CrossRefGoogle Scholar
  45. Kinsara A, Patnaik SN, Cocking EC, Power JB (1986) Somatic hybrid plants of Lycopersicon esculentum Mill. and Lycopersicon peruvianum Mill. J Plant Physiol 125: 225–234Google Scholar
  46. Kohn H, Schieder R, Schieder O (1985) Somatic hybrids in tobacco mediated by electrofusion. Plant Sci 38: 121–128CrossRefGoogle Scholar
  47. Krumbiegel G, Schieder O (1979) Selection of somatic hybrids after fusion of protoplats from Datura innoxia and Atropa belladonna. Planta 145: 371–375CrossRefGoogle Scholar
  48. Kumashiro T, Kubo T (1986)a Cytoplasm transfer of Nicotiana deberyii to N. tabacum by protoplast fusion. Jap J Breed 36: 39–48Google Scholar
  49. Kumashiro T, Kubo T (1986)b Stability of agronomic traits in cytoplasmic male sterile tobacco obtained by protoplast fusion. Jap J Breed 36: 284–290Google Scholar
  50. Maliga P, Sz-Breznovits A, Marton L (1973) Streptomycin resistant plant from callus culture of haploid tobacco. Nature New Biol 244: 29–30PubMedGoogle Scholar
  51. Medgyesy P, Fejes E, Maliga P (1985) Interspecific chloroplast recombination in a Nicotiana somatic hybrid. Proc Natl Acad Sci 82: 6960–6964PubMedCrossRefGoogle Scholar
  52. Melchers G, Sacristan MD, Holder AA (1978) Somatic hybrid plants of potato and tomato regenerated from fused protoplasts. Carlsberg Res Commun 43: 203–218CrossRefGoogle Scholar
  53. Menczel L, Galiba G, Nagy F, Maliga P (1982) Effects of radiation dosage on efficiency of chloroplast transfer by protoplast fusion in Nicotiana. Genetics 100: 487–495PubMedGoogle Scholar
  54. Menczel L, Polsby LS, Steinback KE, Maliga P (1986) Fusion mediated transfer of triazine resistant chloroplasts; Characterization of Nicotiana tabacum cybrid plants. Mol Gen Genet 205: 201–205CrossRefGoogle Scholar
  55. Muller AJ (1983) Genetic analysis of nitrate reductase deficient tobacco plants regenerated from mutant cells. Evidence for duplicate structured genes. Mol Gen Genet 192: 253–261CrossRefGoogle Scholar
  56. Negrutiu I, DeBrouwer 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
  57. Nagy F, Torok I, Mafiga P (1981) Extensive rearrangements in the mitochondrail DNA in somatic hybrids of Nicotiana tabacum and Nicotiana knightiana. Mol Gen Genet 183: 437–439CrossRefGoogle Scholar
  58. Pandeya RS, Douglas GC, Keller WA, Setterfield G, Patrick ZA (1986) Somatic hybridization between Nicotiana rustica and N. tabacum; Development of tobacco breeding strains with disease resistance and elevated nicotine content. Z Pflanzenzuchtg 96: 346–352Google Scholar
  59. Patnaik G, Cocking EC, Hamill JD, Pental D (1982) A simple procedure for the manual isolation and identification of plant heterokaryons. Plant Sci Lett 24: 105–110CrossRefGoogle Scholar
  60. Paszkowski J, Shillito RD, Saul M, Mandak V, Hohn T, Hohn B, Potrykus I (1984) Direct gene transfer to plants EMBO J 3: 2717–2722PubMedGoogle Scholar
  61. Pelletier G, Chupeau Y (1984) Plant protoplast fusion and somatic cell genetics. Physiol Veg 22: 377–399Google Scholar
  62. Pelletier G, Primard C, Vedel F, Chetrit P, Remy R, Rousselle P, Renard M (1983) Intergeneric cytoplasmic hybridization in Cruciferae by protoplast fusion. Mol Gen Genet 191: 244–250CrossRefGoogle Scholar
  63. Pental D, Cocking EC (1985) Some theoretical and practical possibilities of plant genetic manipulation using plant protoplasts. Hereditas Suppl 3: 83–92Google Scholar
  64. Pental D, Hamill JD, Cocking EC (1984) Somatic hybridization using a double mutant of Nicotiana tabacum. Heredity 53: 79–83CrossRefGoogle Scholar
  65. Pental D, Hamill JD, Pirrie A, Cocking EC (1986) Somatic hybridization of Nicotiana tabacum and Petunia hybrida. Mol Gen Genet 202: 342–347CrossRefGoogle Scholar
  66. Pirrie A, Power JB (1986) The production of fertile, triploid somatic hybrid plants (Nicotiana glutinosa (n) + N. tabacum (2n)) via gametic somatic protoplast fusion. Theor Appl Genet 72: 48–52CrossRefGoogle Scholar
  67. Rhodes MJC, Robins RJ, Hamill JD, Parr AJ (1986) Potential for the production of biochemicals by plant cell cultures. N Zealand J Technol 2: 59–70Google Scholar
  68. Schiller B, Herrmann RG, Melchers G (1982) Restriction endonuclease analysis of plastid DNA from tomato, potato and some of their somatic hybrids. Mol Gen Genet 186: 453–459CrossRefGoogle Scholar
  69. Scowcroft WR, Larkin PJ (1981) Chloroplast DNA assorts randomly in intraspecific somatic hybrids of Nicotiana debenyii. Theor Appl Genet 60: 179–184CrossRefGoogle Scholar
  70. Shepard JF, Bidney D, Barsby T, Kemble R (1983) Genetic transfer in plants through interspecific protoplast fusion. Science 219: 683–688PubMedCrossRefGoogle Scholar
  71. Sundberg F, Glimelius K (1986) A method for production of interspecific hybrids within Brassiceae via somatic hybridization, using resynthesis of Brassica napus as a model. Plant Sci 43: 155–162CrossRefGoogle Scholar
  72. Tabaeizadeh Z, Perennes C, Bergounioux C (1985) Increasing the variability of Lycopersican peruvianum Mill. by protoplast fusion with Petunia hybrida L.. Plant Cell Rep 4: 7–11CrossRefGoogle Scholar
  73. Thompson JA, Abdullah R, Cocking EC (1986) Protoplast culture of rice (Oryza sativa) using media solidified with agarose. Plant Sci 47: 123–133CrossRefGoogle Scholar
  74. Vienken J, Ganser R, Haupp 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
  75. Wallin A, Glimelius K, Eriksson T (1974) The induction of aggregation and fusion of Daucus carota protoplasts by polyethylene glycol. Z Pflanzenphysiol 74: 64–80Google Scholar
  76. Ward M, Davey MR, Cocking EC, Balls M, Clothier RH, Lucy JA (1980) Animal specific membrane components visualized on the surface of Animal/Plant heterokaryons. Protoplasma 104: 75–80CrossRefGoogle Scholar
  77. Watts JW, King JM (1984) A simple method for large scale electrofusion and culture of plant protoplasts. Bioscience Rep 4: 335–342CrossRefGoogle Scholar
  78. Wetter LR, Kao KN (1980) Chromosomes and isoenzyme studies on cells derived from protoplast fusion of Nicotiana glauca with Glycine maxNicotiana glauca cell hybrids. Theor Appl Genet 57: 273–276CrossRefGoogle Scholar
  79. White DWR, Vasil IK (1979) Use of amino acid analogue resistant cell lines for selection of Nicotiana sylvestris somatic cell hybrids. Theor Appl Genet 55: 107–112CrossRefGoogle Scholar
  80. Watts JW, Doonan J, Cove DJ, King JM (1985) Production of somatic hybrids of moss by electrof usion. Mol Gen Genet 199: 349–351CrossRefGoogle Scholar
  81. Yarrow SA, Wu SC, Barsby TL, Kemble RJ, Shepard JF The introduction of CMS mitochondria to triazine tolerant Brassica napus L. var Regent by micromanipulation of individual heterokaryons. Plant Cell Rep 5: 415–418Google Scholar
  82. Yamada Y, Yank ZQ, Tang DT (1986) Plant regeneration from protoplast derived callus of rice (Oryza sativa L.). Plant Cell Rep 5: 85–88CrossRefGoogle Scholar
  83. Ye J, Hauptmann RM, Smith AG, Widholm JM (1987) Selection of a Nicotiana plumbaginifolia universal hybridizer and its use in intergeneric somatic hybrid formation. Mol Gen Genet 208: 474–480CrossRefGoogle Scholar
  84. Zelcer A, Aviv D, Galun E (1978) Interspecific transfer of cytoplasmic male sterility by fusion between protoplasts of normal Nicotiana sylvestris and X-ray irradiated protoplasts of male sterile N. tabacum. Z Pflanzenphysiol 90: 397–407Google Scholar
  85. Zenkteler M, Melchers G (1978) In vitro hybridization by sexual methods and by fusion of protoplasts. Theor Appl Genet 52: 81–90Google Scholar
  86. Zimmermann U, Scheurich P (1981) High frequency fusion of plant protoplasts by electrical fields. Planta 151: 26–32CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1988

Authors and Affiliations

  • John D. Hamill
    • 1
  • Edward C. Cocking
    • 2
  1. 1.Plant Biotechnology Group, (Norwich Laboratory)Institute of Food ResearchNorwichUK
  2. 2.Dept. of BotanyUniversity of NottinghamNottinghamUK

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