Skip to main content
SpringerLink
Log in
Book cover

Electroporation and Electrofusion in Cell Biology pp 343–354Cite as

Plant Gene Transfer Using Electrofusion and Electroporation

  • Chapter

Abstract

In recent years, the rapidly developing fields of electrofusion of somatic cells and electroporation of selected cell types have become an extremely important tool in the production and modification of hybrid somatic gene pools. In plants, these procedures have thus far been restricted to protoplasts in which the thick secondary cell wall has been stripped off usually through enzymatic digestion of the cellulosic and pectic components of the cell wall. Protoplasts with a diameter of 35–55 μm can be predictably isolated from normal mesophyll leaf tissue or from cell suspension cultures. These cells are ideally suited for the simple, rapid, and consistent procedures that have been developed for both electrofusion and electroporation (Bates et al., 1987). These procedures are very effective for cells of this size and have made electromanipulation of cell membranes the method of choice for gene transfer due to the simplicity, convenience, and ease of operation.

This is a preview of subscription content, log in via an institution.

Chapter
USD   29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   169.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   219.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Afonso, C. L., Harkins, K. R., Thomas-Compton, M. A., Krejci, A. E., and Galbraith, D. W., 1985, Selection of somatic hybrid plants in Nicotiana through fluorescence-activated sorting of protoplasts, Bio/Technology 3:811–816.

    Article  Google Scholar 

  • Akada, S., and Hirai, A., 1983, Studies on the mode of separation of chloroplast genomes in parasexual hybrid calli. II. Heterogeneous distribution of two kinds of chloroplast genomes in hybrid callus, Plant Sci. Lett. 32:95–100.

    Article  CAS  Google Scholar 

  • Barton, K. A., Binns, A. N., Matzke, A. J. M, and Chilton, M. D., 1983, Regeneration of intact tobacco plants containing full length copies of genetically engineered T-DNA, and transmission of T-DNA to Rl progeny, Cell 32:1033–1043.

    Article  PubMed  CAS  Google Scholar 

  • Bates, G. W., Saunders, J. A., and Sowers, A. E., 1987, Electrofusion principles and applications, in: Cell Fusion (A. E. Sowers, ed.), Plenum Press, New York, pp. 367–395.

    Google Scholar 

  • Belliard, G., Vedel, F., and Pelletier, G., 1979, Mitochondrial recombination in cytoplasmic hybrids of Nicotiana tabacum by protoplast fusion, Nature 281:401–403.

    Article  CAS  Google Scholar 

  • Boeshore, M. L., Lifshitz, I., Hanson, M. R., and Izhar, S., 1983, Novel composition of mitochondrial genomes in Petunia somatic hybrids derived from cytoplasmic male sterile and fertile plants, Mol. Gen. Genet. 190:459–467.

    Article  CAS  Google Scholar 

  • Broglie, R., Coruzzi, G., Fraley, R. T., Rogers, S. G., Horsch, R. B., Niedermeyer, J. G., Fink, C. L., Flick, J. S., and Chua, N. H. 1984, Light-regulated expression of a pea ribulose-1, 5-bisphosphate carboxylase small subunit gene in transformed plant cells, Science 224:838–843.

    Article  PubMed  CAS  Google Scholar 

  • Chen, K., Wildman, S. G., and Smith, H. H. 1977, Chloroplast DNA distribution in parasexual hybrids as shown by polypeptide composition of fraction I protein, Proc. Natl. Acad. Sci. USA 74:5109–5112.

    Article  PubMed  CAS  Google Scholar 

  • Chu, G., Hayakawa, H., and Berg, P., 1987, Electroporation for the efficient transfection of mammalian cells with DNA, Nucleic Acids Res. 15:1311–1326.

    Article  PubMed  CAS  Google Scholar 

  • Crossway, A., Oakes, J., Irvine, J., Ward, B., Knauf, V., and Schumaker, C., 1986, Integration of foreign DNA following microinjection of tobacco mesophyll protoplasts, Mol. Gen. Genet. 202:179–185.

    Article  CAS  Google Scholar 

  • Dudits, D., Hadlaczky, G., Levi, E., Fejer, O., Haydu, Z., and Lazar, G., 1977, Somatic hybridization of Daucus carota and D. capillifolius by protoplast fusion, Theor. Appl. Genet. 51:127–132.

    Google Scholar 

  • Evans, D. A., 1983, Protoplast fusion, in: Handbook of Plant Cell Culture, Volume 1 (D. A. Evans, W. R. Sharp, P. V. Ammirato, and Y. Yamada, eds.), Macmillan Co., New York, pp. 291–321.

    Google Scholar 

  • Falkner, F. G., Neumann, E., and Zachau, H. G., 1984, Tissue specificity of the initiation of immunoglobulin κ gene transcription, Z. Physiol. Chem. 365:1331–1343.

    Article  CAS  Google Scholar 

  • Flick, C. E., Kut, S. A., Bravo, J. E., Gleba, Y. Y., and Evans, D. A., 1985, Segregation of organelle traits following protoplast fusion in Nicotiana, Bio/Technology 3:555–560.

    Article  CAS  Google Scholar 

  • Fluhr, R., Aviv, D., Edelman, M, and Galun, E., 1983, Hybrids containing mixed and sorted-out chloroplasts following interspecific somatic fusions in Nicotiana, Theor. Appl. Genet. 65:289–294.

    Article  Google Scholar 

  • 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. USA 80:4803–4807.

    Article  PubMed  CAS  Google Scholar 

  • Fromm, M., Taylor, L. P., and Walbot, V., 1985, Expression of genes transferred into monocot and dicot plant cells by electroporation, Proc. Natl. Acad. Sci. USA 82:5824–5828.

    Article  PubMed  CAS  Google Scholar 

  • Fromm, M. E., Taylor, L. P., and Walbot, V., 1986, Stable transformation of maize after gene transfer by electroporation, Nature 319:791–793.

    Article  PubMed  CAS  Google Scholar 

  • Galbraith, D. W., and Mauch, T. J., 1980, Identification of fusion of plant protoplasts. II. Conditions for the reproducible fluorescence labelling of protoplasts derived from mesophyll tissue, Z. Pflanzenphysiol. 98:129–140.

    Google Scholar 

  • Glimelius, K., and Bonnett, H. T., 1981, Somatic hybridization in Nicotiana: Restoration of photoautotrophy to an albino mutant with defective plastids, Planta 153:497–503.

    Article  Google Scholar 

  • Gupta, P. P., Gupta, M., and Schieder, O., 1982, Correction of nitrate reductase defect in auxotrophic plant cells through protoplast-mediated intergeneric gene transfers, Mol. Gen. Genet. 188:378–383.

    Article  CAS  Google Scholar 

  • Harms, C. T., Potrykus, I., and Widholm, J. M., 1981, Complementation and dominant expression of amino acid analogue resistance markers in somatic hybrid clones from Daucus carota after protoplast fusion, Z. Pflanzenphysiol. 101:377–390.

    CAS  Google Scholar 

  • Harms, C. T., Oertli, J. J., and Widholm, J. M., 1982, Characterization of amino acid analogue resistant somatic hybrid cell lines of Daucus carota L., Z. Pflanzenphysiol. 106:239–249.

    CAS  Google Scholar 

  • Horn, M. E., Kameya, T., Brotherton, J. E., and Widholm, J. M., 1983, The use of amino acid analog resistance and plant regeneration ability to select somatic hybrids between Nicotiana tabacum and N. glutinosa, Mol. Gen. Genet. 192:235–240.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Ito, H., Murata, K., and Kimura, A., 1984, Transformation of intact yeast cells treated with alkali cations or thiol compounds, Agric. Biol. Chem. 48:341–347.

    Article  CAS  Google Scholar 

  • Iwai, S., Nagao, T., Nakata, K., Kawashima, N., and Matsuyama, S., 1980, Expression of nuclear and chloroplastic genes coding for fraction-1 protein in somatic hybrids of Nicotiana tabacum + rustica, Planta 147:414–417.

    CAS  Google Scholar 

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

    Article  PubMed  Google Scholar 

  • Maliga, P., Lazar, G., Joo, F., Nagy, A. H., and Menczel, L., 1977, Restoration of morphogenic potential in Nicotiana by somatic hybridization, Mol. Gen. Genet. 157:291–296.

    Article  CAS  Google Scholar 

  • Marton, L., Sidorov, B., Baisini, G., and Maliga, P., 1982, Complementation in somatic hybrids indicates four types of nitrate reductase deficient lines in Nicotiana plumbaginifolia, Mol. Gen. Genet. 187:1–3.

    Article  CAS  Google Scholar 

  • Matthews, B. F., and Widholm, J. M., 1985, Organelle DNA composition and isoenzyme expression in an interspecific somatic hybrid of Daucus, Mol. Gen. Genet. 198:371–376.

    Article  CAS  Google Scholar 

  • Medgyesy, P., Menczel, L., and Maliga, P., 1980, The use of cytoplasmic streptomycin resistance: Chloroplast transfer from Nicotiana tabacum into Nicotiana sylvestris, and isolation of their somatic hybrids, Mol. Gen. Genet. 179:693–698.

    Article  CAS  Google Scholar 

  • Medgyesy, P., Fejes, E., and Maliga, P., 1985, Interspecific chloroplast recombination in a Nicotiana somatic hybrid, Proc. Natl. Acad. Sci. USA 82:6960–6964.

    Article  PubMed  CAS  Google Scholar 

  • Miller, P. D., and Sinden, S. L., 1986, Effects of protoplast source and buffer on electroporation efficiency, Mid-Atlantic Plant Molecular Biology Society, Proceedings, p. 12.

    Google Scholar 

  • Mischke, S., Saunders, J. A., and Owens, L., 1986, A versatile low-cost apparatus for cell electrofusion and other electrophysiological treatments, J. Biochem. Biophys. Methods 13:65–75.

    Article  PubMed  CAS  Google Scholar 

  • Morikawa, H., Iida, A., Matsuri, C., Ikegami, M., and Yamada, Y., 1986, Gene transfer into intact plant cells by electroinjection through cell walls and membranes, Gene 41:121–124.

    Article  PubMed  CAS  Google Scholar 

  • Nagy, F., Torok, I., and Maliga, P., 1981, Extensive rearrangements in the mitochondrial DNA in somatic hybrids of Nicotiana tabacum and Nicotiana knightiana, Mol. Gen. Genet. 183:437–439.

    Article  CAS  Google Scholar 

  • Neumann, E., Schaefer-Ridder, M., Wang, Y., and Hofschneider, P. H., 1982, Gene transfer into mouse lyoma cells by electroporation in high electric fields, EMBO J. 1:841–845.

    PubMed  CAS  Google Scholar 

  • Nishiguchi, M., Langridge, W. H. R., Szalay, A. A., and Zaitlin, M., 1986, Electroporation-mediated infection of tobacco leaf protoplasts with tobacco mosaic virus RNA and cucumber mosaic virus RNA, Plant Cell Rep. 5:57–60.

    Article  CAS  Google Scholar 

  • Okada, K., Nagata, T., and Takebe, I., 1986a, Introduction of functional RNA into plant protoplasts by electroporation, Plant Cell Physiol. 27:619–626.

    CAS  Google Scholar 

  • Okada, K., Takebe, I., and Nagata, T., 1986b, Expression and integration of genes introduced into highly synchronized plant protoplasts, Mol. Gen. Genet. 205:398–403.

    Article  CAS  Google Scholar 

  • Ou-Lee, T. M., Turgeon, R., and Wu, R., 1986, Expression of a foreign gene linked to either a plant-virus or a Drosophila promoter, after electroporation of protoplasts of rice, wheat, and sorghum, Proc. Natl. Acad. Sci. USA 83:6815–6819.

    Article  PubMed  CAS  Google Scholar 

  • Pietrzak, M., Shillito, R. D., Hohn, T., and Potrykus, I., 1986, Expression in plants of two bacterial resistance genes after protoplast transformation with a new plant expression vector, Nucleic Acids Res. 14:5857–5868.

    Article  PubMed  CAS  Google Scholar 

  • Potter, H., Weir, L., and Leder, P., 1984, Enhancer-dependent expression of human k immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation, Proc. Natl. Acad. Sci. USA 81:7161–7165.

    Article  PubMed  CAS  Google Scholar 

  • 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 hybrids and Petunia parodii. I. Nutrient media and drug sensitivity complementation selection, Plant Sci. Lett. 10:1–6.

    Article  CAS  Google Scholar 

  • Reiss, M., Jarestreboff, M. M., Bertino, J. R., and Narayanan, R., 1986, DNA-mediated gene transfer into epidermal cells using electroporation, Biochem. Biophys. Res. Commun. 137:244–249.

    Article  PubMed  CAS  Google Scholar 

  • Riggs, C. D., and Bates, G. W., 1986, Stable transformation of tobacco by electroporation: Evidence for plasmid concentration, Proc. Natl. Acad. Sci. USA 83:5602–5606.

    Article  PubMed  CAS  Google Scholar 

  • Saul, M. W., and Potrykus, I., 1984, Species-specific repetitive DNA used to identify interspecific somatic hybrids, Plant Cell Rep. 3:65–67.

    Article  CAS  Google Scholar 

  • Saunders, J. A., and Bates, G. W., 1987, Chemically induced fusion of plant protoplasts, in: Cell Fusion (A. E. Sowers, ed.), Plenum Press, New York, pp. 497–520.

    Google Scholar 

  • Saunders, J. A., Roskos, L. E., Mischke, S., Aly, M. A. H., and Owens, L. D., 1986, Behavior and viability of tobacco protoplasts in response to electrofusion parameters, Plant Physiol. 80:117–121.

    Article  PubMed  CAS  Google Scholar 

  • Schocher, R. J., Shillito, R. D., Saul, M. W., Paszkowski, J., and Potrykus, I., 1986, Co-transformation of unlinked foreign genes into plants by direct gene transfer, Bio/Technology 4:1093–1096.

    Article  CAS  Google Scholar 

  • Serpersu, E. H., Kinosita, K., and Tsong, T. Y., 1985. Reversible and irreversible modification of erythrocyte membrane permeability by electric field, Biochim. Biophys. Acta 812:779–785.

    Article  PubMed  CAS  Google Scholar 

  • Shillito, R. D., Saul, M. W., Paszkowski, J., Muller, M., and Potrykus, I., 1985, High efficiency direct gene transfer to plants, Bio/ Technology 3:1099–1103.

    Article  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • Sowers, A. E., and Kapoor, V., 1987, The electrofusion mechanism in erythrocyte ghosts, in: Cell Fusion (A. E. Sowers, ed.), Plenum Press, New York, pp. 397–408.

    Google Scholar 

  • Wetter, L. R., and Kao, K. N., 1980, Chromosome and isoenzyme studies on cells derived from protoplast fusion of Nicotiana glauca with Glycine max-Nicotiana glauca cell hybrids, Theor. Appl. Genet. 57:273–276.

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Paul D. Miller

    Present address: DNA Plant Technology Corporation, Cinnaminson, New Jersey, 08077, USA

Authors and Affiliations

  1. Germplasm Quality and Enhancement Laboratory, USDA, ARS, Beltsville, Maryland, 20705, USA

    James A. Saunders

  2. Plant Molecular Genetics Laboratory, USDA, ARS, Beltsville, Maryland, 20705, USA

    Benjamin F. Matthews

  3. Vegetable Laboratory, USDA, ARS, Beltsville, Maryland, 20705, USA

    Paul D. Miller

Authors
  1. James A. Saunders
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Benjamin F. Matthews
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Paul D. Miller
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of Bielefeld, Bielefeld, FRG

    Eberhard Neumann

  2. American Red Cross, Rockville, Maryland, USA

    Arthur E. Sowers

  3. Science Applications International Corporation, McLean, Virginia, USA

    Carol A. Jordan

Rights and permissions

Reprints and permissions

Copyright information

© 1989 Springer Science+Business Media New York

About this chapter

Cite this chapter

Saunders, J.A., Matthews, B.F., Miller, P.D. (1989). Plant Gene Transfer Using Electrofusion and Electroporation. In: Neumann, E., Sowers, A.E., Jordan, C.A. (eds) Electroporation and Electrofusion in Cell Biology. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-2528-2_22

Download citation

  • DOI: https://doi.org/10.1007/978-1-4899-2528-2_22

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4899-2530-5

  • Online ISBN: 978-1-4899-2528-2

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation