Research | Published:

Genetic Transformation of Banana and Plantain (Musa spp.) via Particle Bombardment

Bio/Technologyvolume 13pages481485 (1995) | Download Citation

Subjects

Abstract

We have developed a simple protocol to allow the production of transgenic banana plants. Foreign genes were delivered into embryogenic suspension cells using accelerated particles coated with DNA. Bombardment parameters were optimized for a modified particle gun resulting in high levels of transient expression of the β-glucuronidase gene in both banana and plantain cells. Bombarded banana cells were selected with hygromycin and regenerated into plants. Molecular and histochemical characterization of transformants revealed the stable integration of the transferred genes into the banana genome.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    FAO 1993. Production yearbook 1992. Food and Agriculture Organization of the United Nations, Rome, Italy.

  2. 2

    Barton, K.A. and Miller, M.J. 1993. Production of Bacillus thuringiensis insecticidal proteins in plants, p. 297–315. In: Transgenic Plants. Vol II. Engineering and utilization. Rung, S.-D. and Wu, R. (Eds.). Academic Press Inc., London, UK.

  3. 3

    Hilder, V.A., Gatehouse, A.M.R. and Boulter, D. 1993. Transgenic plants conferring insect tolerance: protease-inhibitor approach, p. 317–338. In: Transgenic Plants. Vol II. Engineering and utilization. Kung, S.-D. and Wu, R. (Eds.). Academic Press Inc., London, UK.

  4. 4

    Wilson, T.M.A. 1993. Strategies to protect crop plants against viruses: Pathogen-derived resistance blossoms. Proc. Natl. Acad. Sci. USA 90: 3134–3141.

  5. 5

    Cornelissen, B.J.C. and Melchers, L.S. 1993. Strategies for control of fungal diseases with transgenic plants. Plant Physiol. 101: 709–712.

  6. 6

    Cammue, B.P.A., De Bolle, M.F.C., Schoofs, H.M.E., Thevissen, K., Osbom, R.W., Rees, S.B. and Broekaert, W.F. 1994. Gene-encoded antimicrobial peptides from plants, p. 91–106. In: Antimicrobial Peptides. The Ciba Foundation (Ed.). John Wiley & Sons Ltd., Sussex, UK.

  7. 7

    Dhed'a, D., Dumortier, F., Panis, B., Vuylsteke, D. and De Langhe, E. 1991. Plant regeneration in cell suspension cultures of the cooking banana cv. ‘Bluggoe’ (Musa spp., ABB group). Fruits 46: 125–135.

  8. 8

    Sági, L., Remy, S., Panis, B., Swennen, R. and Volckaert, G. 1994. Transient gene expression in electroporated banana protoplasts (Musa spp., cv. ‘Bluggoe,’ ABB group). Plant Cell Reports 13: 262–266.

  9. 9

    Panis, B., Van Wauwe, A. and Swennen, R. 1993. Plant regeneration through direct somatic embryogenesis from protoplasts of banana (Musa spp.). Plant Cell Reports 12: 403–4107.

  10. 10

    Fitch, M.M., Manshardt, R.M., Gonsalves, D., Slightom, J.L. and Sanford, J.C. 1990. Stable transformation of papaya via microprojectile bombardment. Plant Cell Reports 9: 189–194.

  11. 11

    Christou, P. and McCabe, D.E. 1992. Prediction of germ-line transformation events in chimeric R0 transgenic soybean plantlets using tissue-specific expression patterns. The Plant Journal 2: 283–290.

  12. 12

    Takeuchi, Y., Dotson, M. and Keen, N.T. 1992. Plant transformation: a simple particle bombardment device based on flowing helium. Plant Mol. Biol. 18: 835–839.

  13. 13

    Wang, Y-C., Klein, T.M., Fromm, M., Cao, J., Sanford, J.C. and Wu, R. 1988. Transient expression of foreign genes in rice, wheat and soybean cells following particle bombardment. Plant Mol. Biol. 11: 433–439.

  14. 14

    Vain, P., Keen, N., Murillo, J., Radius, C., Nemes, C. and Finer, J.J. 1993. Development of the particle inflow gun. Plant Cell Tissue Organ Culture 33: 237–246.

  15. 15

    Finer, J.J., Vain, P., Jones, M.W. and McMullen, M.D. 1992. Development of the particle inflow gun for DNA delivery to plant cells. Plant Cell Reports 11: 323–328.

  16. 16

    Last, D.I., Brettell, R.I.S., Chamberlain, D.A., Chaudhury, A.M., Larkin, P.J., Marsh, E.L., Peacock, W.J. and Dennis, E.S. 1991. pEmu: an improved promoter for gene expression in cereal cells. Theor. Appl. Genet. 81: 581–588.

  17. 17

    Bekkaoui, F., Datla, R.S.S., Pilon, M., Tautorus, T.E., Crosby, W.L. and Dunstan, D.I. 1990. The effects of promoter on transient expression in conifer cell lines. Theor. Appl. Genet. 79: 353–359.

  18. 18

    Datla, R.S.S., Hammerlindl, J.K., Pelcher, L.E., Crosby, W.L. and Selvaraj, G. 1991. A bifunctional fusion between β-glucuronidase and neomycin phosphotransferase: a broad-spectrum marker enzyme for plants. Gene 101: 239–246.

  19. 19

    Datla, R.S.S., Bekkaoui, F., Hammerlindl, J.K., Pilate, G., Dunstan, D.I. and Crosby, W.L. 1993. Improved high-level constitutive foreign gene expression in plants using an AMV RNA4 untranslated leader sequence. Plant Sci. 94: 139–149.

  20. 20

    Christensen, A.H., Sharrock, R.A. and Quail, P.H. 1992. Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation. Plant Mol. Biol. 18: 675–689.

  21. 21

    Taylor, M.G., Vasil, V. and Vasil, I.K. 1993. Enhanced GUS gene expression in cereal/grass cell suspesions and immature embryos using the maize ubiquitin-based plasmid pAHC25. Plant Cell Reports 12: 491–495.

  22. 22

    Gritz, L. and Davies, J. 1983. Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. Gene 25: 179–188.

  23. 23

    Simmonds, N.W. 1966. Bananas. Longmans, Green and Co. Ltd., London, UK.

  24. 24

    Cammue, B.P.A., De Bolle, M.F.C., Terras, F.R.G., Proost, P., Van Damme, J., Rees, S.B., Vanderleyden, J. and Broekaert, W.F. 1992. Isolation and characterization of a novel class of plant antimicrobial peptides from Mirabilis jalapa L. seeds. J. Biol. Chem. 267: 228–2233.

  25. 25

    Broekaert, W.F., Marien, W., Terras, F.R.G., De Bolle, M.F.C., Proost, P., Van Damme, J., Dillen, L., Claeys, M., Rees, S.B., Vanderleyden, J. and Cammue, B.P.A. 1992. Antimicrobial peptides from Amamrtthus caudatus seeds with sequence homology to the cysteine/glycine-rich domain of chitin-binding proteins. Biochemistry 31: 4308–4314.

  26. 26

    Terras, F.R.G., Schoofs, H.M.E., De Bolle, M.F.C., Van Leuven, F., Rees, S.B., Vanderleyden, J., Cammue, B.P.A. and Broekaert, W.F. 1992. Analysis of two novel classes of plant antifungal proteins from radish (Raphanus sativus L.) seeds. J. Biol. Chem. 267: 15301–15309.

  27. 27

    Cammue, B.P.A., De Bolle, M.F.C., Terras, F.R.G. and Broekaert, W.F. 1993. Fungal disease control in Musa: Application of new antifungal proteins, p. 221–225. In: Breeding Banana and Plantain for Resistance to Diseases and Pests. Ganry, J. (Ed.). CIRAD, Montpellier, France.

  28. 28

    Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497.

  29. 29

    Christou, P., Ford, T.L. and Kofron, M. 1991. Production of transgenic rice (Oryza saliva L.) plants from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos. Bio/Technology 9: 957–962.

  30. 30

    Jobling, S.A. and Gehrke, L. 1987. Enhanced translation of chimaeric messenger RNAs containing a plant viral untranslated leader sequence. Nature 325: 622–625.

  31. 31

    Birnboim, H.C. and Doly, J. 1979. A rapid alkaline lysis procedure for screening recombinant plasmids. Nucl. Acids Res. 7: 1513–1523.

  32. 32

    Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular cloning: a laboratory manual. Second edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.

  33. 33

    Sanford, J.C., Smith, F.D. and Russell, J.A. 1993. Optimizing the biolistic process for different biological applications. Meth. Enzymol. 217: 483–509.

  34. 34

    Jefferson, R.A. 1987. Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5: 387–405.

  35. 35

    Stewart, C.N., and Via, L.E. 1993. A rapid CTAB DNA isolation technique for RAPD fingerprinting and other PCR applications. BioTechniques 14: 748–750.

  36. 36

    Murray, M.G. and Thompson, W.F. 1980. Rapid isolation of high molecular weight plant DNA. Nucl. Acids Res. 8: 4321–4325.

  37. 37

    Zhou, M.Y., Xue, D., Gomez-Sanchez, E.P. and Gomez-Sanchez, C.E. 1994. Improved downward capillary transfer for blotting of DNA and RNA. BioTechniques 16: 58–59.

  38. 38

    Neuhaus-Url, G. and Neuhaus, G. 1993. The use of the nonradioactive digoxigenin chemiluminescent technology for plant genomic Southern blot hybridization: a comparison with radioactivity. Transgenic Res. 2: 115–120.

Download references

Author information

Affiliations

  1. Laboratory of Tropical Crop Husbandry, Catholic University of Leuven, K. Mercierlaan 92, B-3001, Leuven, Belgium

    • László Sági
    • , Bart Panis
    • , Serge Remy
    • , Hilde Schoofs
    • , Kris De Smet
    •  & Rony Swennen
  2. F.A. Janssens Laboratory of Genetics, Catholic University of Leuven, W. de Croylaan 42, B-3001, Leuven, Belgium

    • Bruno P.A. Cammue

Authors

  1. Search for László Sági in:

  2. Search for Bart Panis in:

  3. Search for Serge Remy in:

  4. Search for Hilde Schoofs in:

  5. Search for Kris De Smet in:

  6. Search for Rony Swennen in:

  7. Search for Bruno P.A. Cammue in:

Corresponding author

Correspondence to László Sági.

About this article

Publication history

Received

Accepted

Issue Date

DOI

https://doi.org/10.1038/nbt0595-481

Further reading