1. Grow healthy Camelina plants for about 2-3 week.

2. Clip main shoots to encourage proliferation of many secondary branches.

3. Waiting until plants have flowers

4. Prepare Agrobacterium tumefaciens strain carrying gene of interest on a binary vector. Grow a large liquid culture @ 28-30^oC in LB with antibiotics to select for the binary plasmid, or grow in other media.

5. Spin down Agrobacterium, resuspend to OD₆₀₀ = 0.8 (can be higher or lower) in 5% Sucrose solution (made fresh is better, no need to autoclave). Add Silwet L-77 to a concentration of 0.05% (500 ul/L) and mix well. You will need 400-500 ml for 6-9 3.5" (9cm) pots.

6. Dip flower parts of plant in Agrobacterium solution for 2 to 3 minutes in vaccum.

7. Place dipped plants laid down on a tray and cover for 16 to 24 hours to maintain high humidity.

8. Tie plants in the pot with a stick as long as plant high. Water and grow plants normally. Stop watering as seeds become mature. Do not water on the top part (seed and flower)

9. Harvest dry seed. Transformants are usually all independent, but are guaranteed to be independent if they come off of separate plants.

10. Select for transformants using fluorescence or herbicide (basta) selectable marker.

11. Put the transgenic seed (color) to soil for next generation.

12. Look at transgenic seed with red color

Transgenic seeds (left) and WT seeds (right). It was took in regular light and with out any filter by Kodak camera

Video clip for Camelina transformation

References:

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 Sciences 316:1194-1199.