Agrobacterium-mediated transformation of Ruta graveolens L.

Karine Lièvre (1) , Thi Le Minh Tran (1) , Sébastien Doerper (2) , Alain Hehn (2) , Paul Lacoste (1) , Brigitte Thomasset (2) , Frederic Bourgaud (2) , Eric Gontier (2)

1 INRA - Institut National de la Recherche Agronomique
2 LAE - Laboratoire Agronomie et Environnement

Agrobacterium tumefaciens is used to develop a genetic transformation method for a medicinal plant Ruta graveolens. The direct plant regeneration strategy is preferred to callus line establishment. In vitro seedlings, 2- -to 3-wk-old, are used to excise hypocotyls and co-cultivated for 3 d with A. tumefaciens strain C58C1Rif containing plasmid pTDE4 harbouring neomycin phosphotransferase (npt II, kanamycin resistance) and beta-glucuronidase encoding genes. The Southern blot analysis has shown that 78% kanamycin resistant plants contain gene encoding beta-glucuronidase. The GUS histochemical assay shows that 67% transgenic plants exhibit the corresponding enzymatic activity. Routine transformation efficiency of R. graveolens L. is 11% and could reach up to 22%. Transgenic plants are grown in the greenhouse within 4 months after the initial seedlings.

Domaines: Sciences du Vivant [q-bio]; Sciences de l'environnement

Karine Lièvre, Thi Le Minh Tran, Sébastien Doerper, Alain Hehn, Paul Lacoste, et al.. Agrobacterium-mediated transformation of Ruta graveolens L.. Protocols for In Vitro Cultures and Secondary Metabolite Analysis of Aromatic and Medicinal Plants, 547, Humana Press, 411 p., 2009, Methods in Molecular Biology, 978-1-60327-286-5 978-1-60327-287-2. ⟨10.1007/978-1-60327-287-2_19⟩. ⟨hal-02822605⟩