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Génomique fonctionnelle des phytopathosystèmes

Agrobacterium-mediated floral transformation
Floral dipping

This protocol is derived from:

Clough, S.J., and Bent, A.F. (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. The Plant Journal 16, 735-743.

however we observed that transformation efficiency could be increased when Silwet L-77 is substituted by OFX-309

Mireault, C., Paris, L.-P., Germain, H. 2014. Enhancement of the Arabidopsis floral dip method with XIAMETER OFX-0309 as alternative to Silwet L-77 surfactant. Botany. 92:1-3

YEB medium

5g/l beef-extract
1g/l yeast-extract
5g/l peptone
5g/l sucrose
2 mM MgSO4


  1. Inoculate colony in 4 ml culture in YEB media with appropriate antibiotics. Shake O/N @ 28C
  2. Use 1 ml to start large 300 mL of the same media with same antibiotic, again O/N @ 28C
  3. Spin the cells at 5000 rpm for 10 minutes
  4. Discard supernatant
  5. Resuspend cells in 300 mL 5% sucrose, 0.05% OFX-309 in water.
  6. Adjust OD600 at 0.8-1.5 in 200-300ml
  7. Dip or spray the flowers of 4-5 weeks plants (stems half of full size is better than full size plants), cover with a dome. Crack dome after 24h, remove after 48h.

Tips to improve efficiency

  1. Cut the main stem of the plants a few days after bolting to allow several axillary shoots to come emerge. This will cause a little delay but more flowers will be available for transformation.
  2. Re-transform 1 week later, can also be done a third time if plants still look healthy.
  3. If using a lousy selection like hygromycin or kanamycin (by opposition to Basta) all siliques can be removed from plants prior to transformation to decrease the number of seeds to be plated.