Advertisement

Electroporation of Brassica

Protocol
  • 677 Downloads
Part of the Methods in Molecular Biology™ book series (MIMB, volume 55)

Abstract

A main objective of electroporation is to transform cells by stable incorporation and expression of foreign genetic material. Incorporation may result in transformation with marker genes, as well as desired traits from a breeder’s point of view (e.g., male sterility, virus resistance, insect resistance). By using electroporation, a variety of plant species refractory to other transformation techniques have been transformed.

Keywords

Male Sterility Cytoplasmic Male Sterility Transformation Frequency NPTII Gene Electroporation Buffer 
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.
This is a preview of subscription content, log in to check access.

References

  1. 1.
    Eimert, K., Schroder, C., and Siegemund, F. (1992) Expression of the NPTII-sequence in cauliflower after injection of agrobacteria into seeds. J. Plant Physiol. 140, 37–40.Google Scholar
  2. 2.
    Charest, P. J., Holbrook, L. A., Gabard, J., Iyer, V. N., and Miki, B. L. (1988) Agrobacterium mediated transformation of thin layer explants from Brassica napus. L. Theor. Appl. Genet. 75, 438–445.CrossRefGoogle Scholar
  3. 3.
    Peecham, P. M. (1988) Successful cocultivation of Brassica napus microspores and proembryos with Agrobacterium. Plant Cell Rep. 8, 387–390.CrossRefGoogle Scholar
  4. 4.
    Bergman, P. and Glimelius, K. (1993) Electroporation of rapeseed protoplasts transient and stable transformation. Physiol. Plant 88, 604–611.CrossRefGoogle Scholar
  5. 5.
    Guerche, P., De Almeida, E. R. P., Schwarztein, M. A., Cander, E., Krebbers, E., and Pelletier, G. (1990) Expression of the S2 albumin from Bertholletia excelsa in Brassica napus. Mol. Gen. Genet. 221, 306–314.PubMedCrossRefGoogle Scholar
  6. 6.
    Herve, C., Rouan, D., Guerche, P., Montane, M.-H., and Yot, P. (1993) Molecular analysis of transgenic rapeseed plants obtained by direct transfer of two separate plasmids containing, respectively, the cauliflower mosaic virus coat protein and a selectable marker gene. Plant Sci. 91, 181–193.CrossRefGoogle Scholar
  7. 7.
    Boyer, J.-C., Zaccomer, B., and Haenni, A.-L. (1993) Electrotransfection of turnip yellow mosaic virus RNA into Brassica leaf protoplasts and detection of viral RNA products with non-radioactive probe. J. Gen. Virol. 74, 1911–1917.PubMedCrossRefGoogle Scholar
  8. 8.
    Eimert, K. and Siegemund, F. (1992) Transformation of cauliflower (Brassica oleracea L. var. botrytis) an experimental survey. Plant Mol. Biol. 19, 485–490.PubMedCrossRefGoogle Scholar
  9. 9.
    Jefferson, R. A. (1987) Assaying chimeric genes in plants the GUS gene fusion system. Plant Mol. Biol. Rep. 5, 387–405.CrossRefGoogle Scholar
  10. 10.
    Jefferson, R. A., Kavanagh, T. A., and Bevan, M. W. (1987) GUS fusions. β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6, 3901–3907.PubMedGoogle Scholar
  11. 11.
    Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol. Plant 15, 473–479.CrossRefGoogle Scholar
  12. 12.
    Menczel, L. and Wolfe, K. (1984) High frequency of fusion induced in freely suspended protoplast mixtures by polyethylene glycol and dimethylsulfoxide at high pH. Plant Cell Rep. 3, 196–198.CrossRefGoogle Scholar
  13. 13.
    Kao, K. N., Constabel, F., Michayluk, M. R., and Gamborg, O. L. (1974) Plant protoplast fusion and growth of intergeneric hybrid cells. Planta. 120, 215–227.CrossRefGoogle Scholar
  14. 14.
    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
  15. 15.
    Siegemund, F. and Tschuch, G. (1984) Elektrisch induzierte Fusion von Lycopersicon-Protoplasten unterschiedlicher Herkunft. Arch. Zúchtungsforsch 14, 163–168.Google Scholar
  16. 16.
    Negrutiu, I., Shillito, R., Potrykus, I., Biasini, G., and Sala, F. (1987) Hybrid genes in the analysis of transformation conditions. I. Setting up a simple method for direct gene transfer in plant protoplasts. Plant Mol. Biol. 8, 363–373.CrossRefGoogle Scholar
  17. 17.
    Guerche, P., Charbonmer, M., Jounanin, J., Tourneur, C., Paszkowski, J., and Pelletier, G. (1987) Direct gene transfer by electroporation in Brassica nupus. Plant Sci. 52, 111–116.CrossRefGoogle Scholar
  18. 18.
    McDonnell, R. E., Clark, R. D., Smith, W. A., and Hinchee, M. A. (1987) A simplified method for detection of neomycm phosphotransferaseII activity in transformed plant tissues. Plant Mol. Biol. Rep. 5, 380–386.CrossRefGoogle Scholar
  19. 19.
    Junghans, H. and Metzlaff, M. (1990) A simple and rapid method for the preparation of total plant DNA. BioTechniques 8, 176.PubMedGoogle Scholar
  20. 20.
    Sambrook, J., Fritsch, E. F., and Maniatis, T. (eds.) (1982) Molecular Cloning, A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
  21. 21.
    Raeder, U. and Broda, P. (1984) Comparisons of the lignin-degrading white rot fungi Phanerochaete orysosporum and Sprototrichum pulverutentum at the DNA level. Curr. Genet. 8, 499–506.CrossRefGoogle Scholar
  22. 22.
    Shillito, R. D., Saul, M. W., Paszkowski, J., Müller, M., and Potrykus, I. (1985) High efficiency direct gene transfer to plants. BioTechnology 3, 1099–1103.CrossRefGoogle Scholar
  23. 23.
    Rhodes, C. A., Pierce, D. A., Mettler, I. J., Mascarenhas, D., and Detmer, J. J. (1988) Genetically transformed maize plants from protoplasts. Science. 240, 204–207.PubMedCrossRefGoogle Scholar
  24. 24.
    De Block, M., De Brouwer, D. D., and Tenning, P. (1989) Transformation of Brassica napus and Brassica oleracea using Agrobacterium tumefaciens and the expression of the bar and neo genes in the transgenic plants. Plant Physiol. 91, 694–701.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1995

Authors and Affiliations

  1. 1.Department of GeneticsUniversity of HalleSaaleGermany

Personalised recommendations

Cite protocol

Buy options