Skip to main content
SpringerLink
Log in

Transient foreign gene expression in polyethylene/glycol treated or electropulsated Eucalyptus gunnii protoplasts

  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

Conditions for optimal transient gene expression of reporter genes (chloramphenicol acetyl transferase and β-glucuronidase) by polyethylene glycol or electrical treatment of Eucalyptus gunnii protoplasts derived from callus or cell suspension cultures were investigated. The effciency of electropermeabilisation depended on several factors including electrical parameters, pH and the source of protoplasts. Polyethylene glycol mediated DNA uptake was highly stimulated by heat shock pretreatment and was also more efficient than the electrical treatment. For both treatments the nature of the promoter associated with the reporter gene was very important. A promoter corresponding to a protein synthesis elongation factor gene was more effective than the classical promoter of the 35 S transcript from cauliflower mosaic virus.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

CaMV:

Cauliflower Mosaic Virus

CAT:

Chloramphenicol Acetyl Transferase

FDA:

Fluorescein Diacetate

GUS:

β Glucuronidase

HS:

Heat Shock

MES:

Morpholino Ethane Sulfonic Acid

MU:

Methyl Umbelliferone

MUG:

Methyl Umbelliferyl Glucuronide

NOS:

Nopaline Synthase

References

  • Adam S (1989) Etude de la sensibilité d'Eucalyptus gunnii à Agrobacterium rhizogenes et A. tumefaciens; facteurs impliqués dans l'obtention de réponses de transformation et problèmes liés à la régénération de plantes entières transformées. Thèse de Doctorat de l'Université Paris VI

  • Ahuja MR (1988) Gene transfer in forest trees. In: Hanover JW & Keathley DE (Eds) Genetic Manipulation of Woody Plants (pp 25–41)

  • Axelos M, Bardet C, Liboz T, Le Van Thai A, Curie C & Lescure B (1989) The gene family encoding the Arabidopsis thaliana translation elongation factor EF-1 α: Molecular cloning characterization and expression. Mol. Gen. Genetic. 219: 106–112

    Google Scholar 

  • Bekkaoui F, Datla RSS, Pilon M, Tautorus TE, Crosby WL & Dunstan DI (1990) The effects of promoter on transient expression in conifer cell lines. Theor. Appl. Genet. 79: 353–359

    Google Scholar 

  • Bekkaoui F, Pilon M, Laine E, Raju DSS, Crosby WL & Dunstan DI (1988) Transient gene expression in electroporated Picea glauca protoplasts. Plant Cell Rep. 7: 481–484

    Article  Google Scholar 

  • Chupeau Y, Missonier C, Hommel MC & Goujaud J (1978) Somatic hybrids of plants by fusion of protoplasts. Mol. Gen. Genet. 165: 239–245

    Article  Google Scholar 

  • Fillati IJ, Sellmer J, Mac Cown B, Maissig B & Comai (1987) Agrobacterium-mediated transformation and regeneration of Populus. Mol. Gen. Genet. 206: 192–199

    Article  Google Scholar 

  • Fromm M, Taylor LP & Walbot V (1985) Expression of genes electroporated into monocot and dicot plant cells. Proc. Natl. Acad. Sci. USA. 82: 5824–5828

    PubMed  Google Scholar 

  • Gorman C (1985) High efficiency gene transfer into mammalian cells. In: Glever DM (Ed) DNA Cloning. A Practical Approach, Vol II. Irl Press

  • Hauptmann RM, Ozias-Akins P, Vasil V, Tabacizadeh Z, Rogers Sg, Horsch RB, Vasil IK & Fraley RT (1988) Transient expression of electroporated DNA in monocotyledonous and dicotyledonous species. Plant Cell Rep. 6: 265–270

    Article  Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: The GUS gene fusion system. Plant Molec. Biol. Rep. 5: 387–405

    Google Scholar 

  • Lockwood GB (1984) Alkaloïds of cell suspensions derived from four Papaver spp. and the effect of temperature stress. Z. Pflanzenphysiol. 114: 361–363

    Google Scholar 

  • Mac Granahan GH, Leslie CA, Uratsu SL, Martin LA & Dandekar AM (1988) Agrobacterium-mediated transformation of walnut somatic embryos and regeneration of transgenic pl walnut somatic embryos and regeneration of transgenic plants. Biotechnology 6: 800–804

    Article  Google Scholar 

  • Montané MH (1989) Approche biophysiochimique de la transformation directe de protoplastes végétaux par des champs électriques: cas du tabac. Thèse de Doctorat de l'Institut National Polytechnique de Toulouse. July 1989

  • Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–477

    Google Scholar 

  • Negrutiu I, Shillito R, Potrykus I, Biasini G & 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 Molec. Biol. 8: 363–373

    Google Scholar 

  • Pietrzack M, Shillito RD, Hohn T & Potrykus I (1986) Expression in plants of two bacterial antibiotic resistance genes after protoplast transformation with a new plant expression vector. Nucleic Acids Res. 14: 5857–5868

    PubMed  Google Scholar 

  • Prols M, Topfer R, Schell J & Steinbib H (1988) Transient gene expression in tobacco protoplasts: I-time course of CAT apperance. Plant Cell Rep. 7: 221–224

    Google Scholar 

  • Rols MP & Teissie J (1987) Ionic strength modulation of electrically induced permeabilization and associated fusion of mammalian cells. Eur. J. Biochem. 179: 109–115

    Google Scholar 

  • Seguin A & Lalonde M (1988) Gene transfer by electroporation in Betulaceae protoplasts: Alnus incana. Plant Cell Rep. 7: 367–370

    Google Scholar 

  • Tautorus TE, Bekkaoui F, Pilon M, Datla RSS, Crosby WL, Fowke LC & Dunstan DI (1989) Factors affecting transient gene expression in electroporated black spruce (Picea mariana) and jack pine (Pinus banksiana) protoplasts. Theor. Genet. 78: 531–536

    Article  Google Scholar 

  • Teulières C, Feuillet C & Boudet AM (1989) Differential characteistics of cell suspension cultures initiated from Eucalyptus gunnii clones differing by their frost tolerance. Plant Cell Rep. 8: 407–410

    Article  Google Scholar 

  • Teulières C & Boudet AM (1991) Isolation and culture of Eucalypt protoplasts: Approaches for improvement of cell sivision frequency and microcallus obtention. Plant Cell Tiss. Org. Cult. (submitted)

  • Visil V, Hauptmann M, Morrish FM & Vasil IK (1988) Comparitive 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  Google Scholar 

  • Widholm JM (1972) The use of fluorescein diacetate and phenosafranine for determining viability of cultured cells. Stain Technol. 47: 189–194

    PubMed  Google Scholar 

  • Zhang HM, Yang H, Rech EL, Golds TJ, Davis AJ, Mulligan BJ, Cocking EC & Davey MR (1988) Transgenic rice plants produced by electroporation-mediated plasmid uptake into protoplasts. Plant Cell Rep. 7: 379–384

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Centre de Physiologie Végétale de, l'Université Paul Sabatier URA CNRS 241, 118 route de Narbonne, 31062, Toulouse Cédex, France

    C. Teulières, J. Grima-Pettenati, C. Curie & A. M. Boudet

  2. Centre de Biochimie et de Génétique Cellulaires du CNRS, Université Paul Sabatier, 118 route de Narbonne, 31062, Toulouse Cédex, France

    J. Teissie

Authors
  1. C. Teulières
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Grima-Pettenati
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Curie
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. Teissie
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. M. Boudet
    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

Teulières, C., Grima-Pettenati, J., Curie, C. et al. Transient foreign gene expression in polyethylene/glycol treated or electropulsated Eucalyptus gunnii protoplasts. Plant Cell Tiss Organ Cult 25, 125–132 (1991). https://doi.org/10.1007/BF00042183

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00042183

Key words

Search

Navigation