Skip to main content
Log in

Germ-line transformation of Arabidopsis lasiocarpa

  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

In planta transformation methods have opened up the possibility of transforming plant species for which no regeneration protocols currently exist. In this study, the suitability of the germ-line transformation method developed for Arabidopsis thaliana was examined for four taxa in the Brassicaceae that have not been previously transformed: Arabidopsis griffithiana, Arabidopsis lasiocarpa, Arabidopsis petraea and Capsella bursa-pastoris. Numerous transformants were obtained for A. lasiocarpa. Transformation of A. lasiocarpa was confirmed at the phenotypic and molecular levels for stably transformed lines and for backcrossed lines segregating the T-DNA insert. Parameters affecting transformation efficiency of A. lasiocarpa were also explored. As with A. thaliana, sucrose and surfactant in the inoculation medium are required for high levels of transformation, although the suitable concentrations of these are different for A. lasiocarpa. Other components present in earlier versions of the inoculation medium had little effect on transformation efficiency. Vacuum infiltration (rather than simple floral dipping) led to higher rates of transformation and did not seriously affect seed production in A. lasiocarpa. Identification of species susceptible to germ-line transformation will aid in determining the factors important for applying this technology to more recalcitrant species.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA et al. (1995) Current Protocols in Molecular Biology, 3rd edition. Wiley, New York.

    Google Scholar 

  • Bechtold N, Ellis J and Pelletier G (1993) In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. C R Acad Sci Paris, Life Sci 316: 1194–1199.

    Google Scholar 

  • Bechtold N and Pelletier G (1998) In planta Agrobacteriummediated transformation of adult Arabidopsis plants by vacuum infiltration. In: Martinez-Zapater J and Salina K (eds.) Methods in Molecular Biology, Arabidopsis Protocols, Vol. 82 (pp. 259–266) Humana Press, Totowa NJ.

    Google Scholar 

  • Birch RG (1997) Plant transformation: problems and strategies for practical application. Ann Rev Plant Phys Plant Mol Biol 48: 297–326.

    Google Scholar 

  • Chang SS, Park SK, Kim BC, Kang BJ, Kim DU and Nam HG (1994) Stable genetic transformation of Arabidopsis thaliana by Agrobacterium inoculation in planta. Plant J 5: 551–558.

    Google Scholar 

  • Church GM and Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci (USA) 81: 1991–1995.

    Google Scholar 

  • Clough SJ and Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16: 735–743.

    Google Scholar 

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

    Google Scholar 

  • Feldman KA and Marks MD (1987) Agrobacterium mediated transformation of germinating seeds of Arabidopsis thaliana: a non-tissue culture approach. Mol Gen Genet 208: 1–9.

    Google Scholar 

  • Haseloff J, Siemering KR, Prasher DC and Hodge S (1997) Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. Proc Natl Acad Sci USA 94: 2122–2127.

    Google Scholar 

  • Katavic V, Haughn GW, Reed D, Marin M and Kunst L (1994) In planta transformation of Arabidopsis thaliana. Mol Gen Genet 245: 363–370.

    Google Scholar 

  • Koorneef M, Alonso-Blanco C and Stam P (1998) Genetic Analysis. In: Martinez-Zapater J and Salina K (ed.), Methods in Molecular Biology: Arabidopsis Protocols, Vol. 82 (pp. 105–117) Humana Press, Totowa, NJ.

    Google Scholar 

  • Mysore KS, Kumar CTR and Gelvin SB (2000) Arabidopsis ecotypes and mutants that are recalcitrant to Agrobacterium root transformation are susceptible to germ-line transformation. Plant J 21: 9–16.

    Google Scholar 

  • O'Kane SL, Schaal BA and Al-Shebaz IA (1996) The origins of Arabidopsis suecica (Brassicaceae) as indicated by nuclear rDNA sequences. Syst Bot 21: 559–566.

    Google Scholar 

  • O'Kane SLJ and Al-Shehbaz IA (1997) A synopsis of Arabidopsis (Brassicaceae). Novon 7: 323–327.

    Google Scholar 

  • Pigliucci M (1998) Ecological and evolutionary genetics of Arabidopsis. Trends Plant Sci 3: 485–489.

    Google Scholar 

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

    Google Scholar 

  • Price, RA Palmer JD and Al-Shehbaz IA (1994) Systematic relationships of Arabidopsis: a molecular and morphological perspective. In: Meyerowitz EM and Somerville CR (eds.) Arabidopsis (pp. 7–10) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  • Qing CM, Fan L, Lei Y, Bouchez D, Tourneur C, Yan L et al. (2000) Transformation of Pakchoi (Brassica rapa L. ssp chinensis) by Agrobacterium infiltration. Mol Breed 6: 67–72.

    Google Scholar 

  • Schulz R and Jensen WA (1968) Capsella embryogenesis: the egg, zygote and young embryo. Am J Bot 36: 629–641.

    Google Scholar 

  • Trieu AT, Burleigh SH, Kardailsky IV, Maldonado-Mendoza IE, Versaw WK, Blaylock LA et al. (2000) Transformation of Medicago truncatula via infiltration of seedlings or flowering plants with Agrobacterium. Plant J 22: 531–541.

    Google Scholar 

  • Ye GN, Stone D, Pang SZ, Creely W, Gonzalez K and Hinchee M (1999) Arabidopsis ovule is the target for Agrobacterium in planta vacuum infiltration transformation. Plant J 19: 249–257.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tague, B.W. Germ-line transformation of Arabidopsis lasiocarpa . Transgenic Res 10, 259–267 (2001). https://doi.org/10.1023/A:1016633617908

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1016633617908

Navigation