Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Paper
  • Published:

Transgenic Plants of Tall Fescue (Festuca arundinacea Schreb.) Obtained by Direct Gene Transfer to Protoplasts

Bio/Technology volume 10, pages 691–696 (1992)Cite this article

Abstract

Chimeric hygromycin phosphotransferase (hph) and phosphinothricin acetyltransferase (bar) genes were introduced, using polyethylene glycol treatment, into protoplasts isolated from embryogenie cell suspension cultures of tall fescue (Festuca arundinacea Schreb.), a graminaceous plant that is an important forage crop in temperate pastures. Colonies resistant to either 200 mg/l hygromycin or 100 mg/l phosphinothricin, respectively, were recovered upon selection using bead–type culture systems. Stable integration of the transgenes in the genomes of plants regenerated from resistant callus clones was shown by Southern hybridization analysis. In situ hybridization of a labeled transgene–probe to metaphase chromosomes is shown for one transgenic primary regenerant. Expression of the transgenes in mature plants was demonstrated by HPH enzyme assay or by phosphinothricin–herbicide spraying.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Vasil, I.K. 1988. Progress in the regeneration and genetic manipulation of cereal crops. Bio/Technology 6: 397–402.

    Google Scholar 

  2. Potrykus, I. 1990 Gene transfer to cereals an assessment. Bio/Technology 8: 535–542.

    CAS  Google Scholar 

  3. Vasil, V. and Vasil, I.K. 1980. Isolation and culture of cereal protoplasts II Embryogenesis and plantlet formation from protoplasts of Pennisetum americanum. Theor. Appl. Genet. 56: 97–99.

    Article  CAS  Google Scholar 

  4. Lu, C.Y., Vasil, V. and Vasil, I.K. 1981. Isolation and culture of protoplasts from Panicum maximum]acq (Guinea grass): somatic embryogenesis and plantlet formation. Z. Pflanzenphysiol. 104: 311–318.

    Article  Google Scholar 

  5. Vasil, V., Wang, D.Y. and Vasil, I.K. 1983. Plant regeneration from protoplasts of Napier grass (Pennisetum purpureum Schum.). Z Pflanzenphysiol. 111: 233–239.

    Article  CAS  Google Scholar 

  6. Dalton, S.J. 1988. Plant regeneration from cell suspension protoplasts of Festuca arundinacea Schreb., Lolium perenne L. and L multiflorum Lam. Plant Cell Tissue Organ Culture 12: 137–140.

    Article  Google Scholar 

  7. Dalton, S.J. 1988. Plant regeneration from cell suspension protoplasts of Festuca arundinacea Schreb (tall fescue) and Lolium perenne L. (perennial ryegrass) J. Plant Physiol. 132: 170–175.

    Article  Google Scholar 

  8. Takamizo, T., Suginobu, K. and Ohsugi, R. 1990. Plant regeneration from suspension culture derived photoplasts of tall fescue (Festuca arundinacea Schreb.) of a single genotype. Plant Science 72: 125–131.

    Article  Google Scholar 

  9. Creemers-Molenaar, J. van der Valk, P., Loeffen, J.P.M. and Zaal, M.A.C.M. 1989. Plant regeneration from suspension cultures and protoplasts of Lolium perenne L. Plant Science 63: 167–176.

    Article  Google Scholar 

  10. Horn, M.E., Conger, B.V. and Harms, C.T. 1988. Plant regeneration from protoplasts of embryogenic suspension cultures of orchardgrass (Dactylis glomerata L.) Plant Cell Rep. 7: 371–374.

    Article  CAS  Google Scholar 

  11. van der Valk, P., Zaal, M.A.C.M. and Creemers-Molenaar, J. 1988. Regeneration of albino plantlets from suspension culture derived protoplasts of Poa pratensis L. (Kentucky bluegrass). Euphytica S: 169–176.

    Article  Google Scholar 

  12. Zaghmout, O.M.F. and Torello, W.A. 1990. Isolation and culture of protoplasts from embryogenic suspension cultures of red fescue (Festuca rubra L.). Plant Cell Rep. 9: 340–343.

    Article  CAS  Google Scholar 

  13. Hauptmann, R.M., Ozias Atkins, P., Vasil, V., Tabaeizadeh, Z., Rogers, S.G., Horsch, R.B., Vasil, I.K. and Fraley, R.T. 1987. Transient expression of electroporated DNA in monocotyledonous and dicotyledonous species. Plant Cell Rep. 6: 265–270.

    Article  CAS  Google Scholar 

  14. Vasil, V., Hauptmann, R.M., Morrish, F.M. and Vasil, J.K. 1988. Comparative analysis of free DNA delivery and expression into protoplasts of Panicum maximum Jacq. (Guinea grass) by electroporation and polyethylene glycol. Plant Cell Rep. 7: 499–503.

    Article  CAS  Google Scholar 

  15. Potrykus, I., Saul, M.W., Petruska, J., Paszkowski, J. and Shillito, R. 1985. Direct gene transfer to cells of a graminaceous monocot. Mol. Gen. Genet. 199: 183–188.

    Article  CAS  Google Scholar 

  16. Hauptmann, R.M., Vasil, V., Ozias-Atkins, P., Tabaeizadeh, Z., Rogers, S.G., Fraley, R.T., Horsch R.B. and Vasil, I.K. 1988. Evaluation of selectable markers for obtaining stable transformants in the Grammeae. Plant Physiol. 86: 602–606.

    Article  CAS  Google Scholar 

  17. Horn, M.E., Shillito, R.D., Conger, B.V. and Harms, C.T. 1988. Transgenic plants of Orchardgrass (Dactylis glomerata L.) from protoplasts. Plant Cell Rep. 7: 469–472.

    Article  CAS  Google Scholar 

  18. Potrykus, I. 1991. Gene transfer to plants assessment of published approaches and results. Ann. Rev. Plant Physiol. Plant. Mol. Biol. 42: 205–225.

    Article  CAS  Google Scholar 

  19. De Block, M., Botterman, J., Vandwiele, M., Dockx, J., Thoen, C., Gossele, V., Rao Movva, N., Thompson, C., van Montagu, M. and Leemans, J. 1987. Engineering herbicide resistance in plants by expression of a detoxifying enzyme. EMBO J. 9: 2513–2518.

    Article  Google Scholar 

  20. Müller, A.J. and Grafe, R. 1978. Isolation and characterization of cell lines of Nicotiana tabacum lacking nutate reductase. Mol. Gen. Genet. 161: 67–76.

    Article  Google Scholar 

  21. Chen, Y. 1986. Anther and pollen culture of rice, p 1–25. In: Haploids of Higher Plants In Vitro. Hu, H. and Yang, H.Y. (Eds) China Academic Publishers and Springer Verlag, Berlin, Germany.

    Google Scholar 

  22. Bilang, R., Iida, S., Peterhans, A., Potrykus, I. and Paszkowski, J. 1991. The 3′ terminal region of the hygromycin B-resistance gene is important for its activity in Escherichia coli and Nicotiana tabacum. Gene 100: 247–250.

    Article  CAS  Google Scholar 

  23. Donn, G., Nilges, M. and Morocz, S. 1990. Stable transformation of maize with a chimaeric modified phosphinothricin acyltransferase gene from Streptomyces viridochromogens, p 53. In: Progress in Plant Cellular and Molecular Biology. Nijkamp, H.J.J., Van der Plas, L.H.W. and Aartrijk, J. (Eds.). Dordrecht, Kluwer Academic Publishers, Dordrecht, The Netherlands.

    Google Scholar 

  24. Zhang, W., McElroy, D. and Wu, R. 1991. Analysis of rice actl 5′ region activity in transgenic rice plants. The Plant Cell 3: 1155–1165.

    CAS  PubMed  PubMed Central  Google Scholar 

  25. McElroy, D., Blowers, A.D., Jenes, B. and Wu, R. 1991. Construction of expression vectors based on the rice actin 1 (Actl) 5′ region for use in monocot transformation. Mol. Gen. Genet. 231: 150–160.

    Article  CAS  Google Scholar 

  26. Takamizo, T., Spangenberg, G., Suginobu, K. and Potrykus, I. 1991. Intergeneric somatic hybridization in Grammeae: Somatic hybrid plants between tall fescue (Festuca arundinacea Schreb.) and Italian ryegrass (Lolium multiflorum Lam.) Mol. Gen. Genet. 231: 1–6.

    Article  CAS  Google Scholar 

  27. Menczel, L., Nagy, F., Kiss, Z. and Maliga, P. 1981 Streptomycin resistant and sensitive somatic hybrids of N. tabacum + N. Knightiana: correlation of resistance to N tabacum plastids Theor. Appl. Genet. 59: 191–195.

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  29. Negrutiu, I., Shillito, R., Potrykus, I., Biasini, G. and Sala, F. 1987. Hybrid genes in the analysis of transformation condition Plant. Mol. Biol. 8: 363–373.

    Article  CAS  Google Scholar 

  30. Shillito, R.D., Paszkowski, J., Müller, M. 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.

    Article  CAS  Google Scholar 

  31. Kyozuka, J., Hayashi, Y. and Shimamoto, K. 1987. High frequency plant regeneration from rice protoplasts by novel nuise culture methods. Mol. Gen. Genet. 206: 408–413.

    Article  CAS  Google Scholar 

  32. Lichtenstein, C. and Draper, J. 1985. Genetic engineering of plants, p. 67–119. In: DNA Cloning, vol II Glover. D.M. (Ed) IRL. Press, Washington, DC.

    Google Scholar 

  33. Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning—a Laboratory Manual, Second edition Cold Spring Harbor Laboratory Press, NY.

    Google Scholar 

  34. Femberg, A.P. and Vogelstein, B. 1983. A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132: 6–13.

    Article  Google Scholar 

  35. Dretzen, G. 1981. A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal. Biochem. 112: 295–298.

    Article  CAS  Google Scholar 

  36. Perez-Vicente, R., Petris, L., Osusky, M., Potrykus, I. and Spangenberg, G. 1992. Molecular and cytogenetic characterization of repetitive DNA sequences from Lolium and Festuca: applications in the analysis of Festulolium hybrids. Theor. Appl. Genet. In press.

  37. Cabanes-Bastos, E., Day, A.G. and Lichtenstein, C.P. 1989. A sensitive and simply assay for neomycin phosphotransferase II activity in transgenic tissue. Gene 77: 169–177.

    Article  CAS  Google Scholar 

  38. Spangenberg, G., Freydl, E., Osusky, M., Nagel, J. and Potrykus, I. 1991. Organelle transfer by microfusion of denned protoplast cytoplast pairs. Theor. Appl. Genet. 81: 477–486.

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Tadashi Takamizo

    Present address: National Grassland Research Institute, Senbonmatsu 768, Nishinasunomachi, Tochigi, Japan

  2. German Spangenberg: Corresponding author.

Authors and Affiliations

  1. Institute for Plant Sciences, Swiss Federal Institute of Technology, CH 8092, Zürich, Switzerland

    Zeng-yu Wang, Victor A. Iglesias, Martin Osusky, Jutta Nagel, Ingo Potrykus & German Spangenberg

Authors
  1. Zeng-yu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tadashi Takamizo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Victor A. Iglesias
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Martin Osusky
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jutta Nagel
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ingo Potrykus
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. German Spangenberg
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, Zy., Takamizo, T., Iglesias, V. et al. Transgenic Plants of Tall Fescue (Festuca arundinacea Schreb.) Obtained by Direct Gene Transfer to Protoplasts. Nat Biotechnol 10, 691–696 (1992). https://doi.org/10.1038/nbt0692-691

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0692-691

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing