Skip to main content
SpringerLink
Log in

Improving transformation efficiency ofArabidopsis thaliana by modifying the floral dip method

  • Protocols
  • Published:
Plant Molecular Biology Reporter Aims and scope Submit manuscript

Abstract

Arabidopsis thaliana transformation with the floral dip method was improved by modifying the cell density and mode of application of theAgrobacterium inoculum. Drops of inoculum were applied 4 times to closed flower buds. The inoculum OD600 was increased from 0.8 to more than 2.0. These modifications improved reproducibility and increased transformation efficiencies to 2–3%.

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

GUS:

β-glucuronidase

MS:

Murashige and Skoog

RT-PCR:

reverse transcriptase-polymerase chain reaction

References

  • Bechtold N, Ellis J, and Pelletier G (1993) In plantaAgrobacterium-mediated gene transfer by infiltration of adultArabidopsis thaliana plants. C R Acad Sci Paris Life Sci 316: 1194–9.

    CAS  Google Scholar 

  • Bent AF (2000)Arabidopsis in planta transformation. Uses, mechanisms, and prospects for transformation of other species. Plant Physiol 124: 1540–7.

    Article  PubMed  CAS  Google Scholar 

  • Chang SS, Park SK, Kim BC, Kang BJ, Kim DU, and Nam HG (1994) Stable genetic transformation ofArabidopsis thaliana byAgrobacterium inoculationin planta. Plant J 5: 551–8.

    Article  CAS  Google Scholar 

  • Clough S and Bent AF (1998) Floral dip: a simplified method forAgrobacterium-mediated transformation ofArabidopsis thaliana. Plant J 16: 735–43.

    Article  PubMed  CAS  Google Scholar 

  • Fesfeux C, Clough SJ, and Bent AF (2000) Female reproductive tissues are the primary target ofAgrobacterium-mediated transformation by theArabidopsis floral-dip method. Plant Physiol 123: 895–904.

    Article  Google Scholar 

  • Gallagher SR (1992) Quantitation of GUS activity by fluorometry. In: Gallagher SR (ed), GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression, pp 47–60, Academic Press, San Diego, CA.

    Google Scholar 

  • Herrera-Estrella L, Depicker A, Van Montagu M, and Schell J (1983) Expression of chimaeric genes transferred into plant cells using a Ti- plasmid-derived vector. Nature 303: 209–13.

    Article  CAS  Google Scholar 

  • Katavic V, Haughn, GW, Reed D, Martin M, and Kunst L (1994) In planta transformation ofArabidopsis thaliana. Mol Gen Genet 245: 363–70.

    Article  PubMed  CAS  Google Scholar 

  • Klein TM, Wolf ED, Wu R, and Sanford JC (1987) High velocity microprojectiles for delivering nucleic acids into living cells. Nature 327: 70–3.

    Article  CAS  Google Scholar 

  • Martín T, Wöhner RV, Hummel S, Willmitzer L, and Frommer WB (1992) The GUS reporter system as a tool to study gene expression. In: Gallagher SR (ed), GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression, pp 23–46, Academic Press, San Diego, CA.

    Google Scholar 

  • Martínez-Trujillo M (2002) Análisis funcional de regiones involucradas en la expresión del gen de Sacarosa-Fosfato Sintasa de arroz (Oryza sativa L.). Tesis de Doctorado. CINVESTAV-IPN. Irapuato, Guanajuato, México.

    Google Scholar 

  • Martínez-Trujillo M., Simpson J, and Herrera-Estrella L (2002) Transgenic plants in modern agriculture. J New Seeds 4: 1–23.

    Article  Google Scholar 

  • McBride KE and Summerfelt KR (1990) Improved binary vectors forAgrobacterium-mediated plant transformation. Plant Mol Biol 14: 269–76.

    Article  PubMed  CAS  Google Scholar 

  • Meinke DW, Cherry M, Dean C, Rounsley SD, and Koorneef M (1998)Arabidopsis thaliana: A model plant for genome analysis. Science 282: 662–82.

    Article  PubMed  CAS  Google Scholar 

  • Schuler MA and Zielinski RE (1989) Methods in Plant Molecular Biology, pp 35–58, Academic Press Inc., San Diego, CA.

    Google Scholar 

  • Stomp AM (1992) Histochemical localization of β-glucuronidase. In: Gallagher SR (ed), GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression, pp 103–13, Academic Press, San Diego, CA.

    Google Scholar 

  • Trieu AT, Burleigh SH, Kardailsky IV, Maldonado-Mendoza IE, Versaw WK, Blaylock LA, Shin H, Chiou TJ, Katagi H, Dewbre GR, and others (2000) Transformation ofMedicago truncatula via infiltration of seedlings or flowering plants withAgrobacterium. Plant J 22: 531–41.

    Article  PubMed  CAS  Google Scholar 

  • Valvekens D, Montague MV, and Lijsbettens MV (1988)Agrobacterium tumefaciens mediated transformation ofArabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci USA 85: 5536–40.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Facultad de Biología, Universidad Michoacana de San Nicolás de Hidalgo, Ciudad Universitaria, 58066, Morelia, Michoacán, México

    Miguel Martinez-Trujillo

  2. Departamento de Ingeniería Genética de Plantas, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Unidad Irapuato, Km 9.6 carretera Irapuato-León, 36500, Irapuato, Guanajuato, México

    Veronica Limones-Briones, José Luis Cabrera-Ponce & Luis Herrera-Estrella

Authors
  1. Miguel Martinez-Trujillo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Veronica Limones-Briones
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. José Luis Cabrera-Ponce
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Luis Herrera-Estrella
    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

Martinez-Trujillo, M., Limones-Briones, V., Cabrera-Ponce, J.L. et al. Improving transformation efficiency ofArabidopsis thaliana by modifying the floral dip method. Plant Mol Biol Rep 22, 63–70 (2004). https://doi.org/10.1007/BF02773350

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02773350

Key words

Search

Navigation