title: Agrobacterium-mediated transformation of Ruta graveolens L.
abstract: 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.
authors.affiliation: UMR Nancy-Université-INRA Agronomie et Environnement Nancy-Colmar, Vandoeuvre-Lès-Nancy, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Lièvre
authors.forename: Thi Lê Minh
authors.initials: TL
authors.lastname: Tran
authors.forename: Sébastien
authors.initials: S
authors.lastname: Doerper
authors.forename: Alain
authors.initials: A
authors.lastname: Hehn
authors.forename: Paul
authors.initials: P
authors.lastname: Lacoste
authors.forename: Brigitte
authors.initials: B
authors.lastname: Thomasset
authors.forename: Frédéric
authors.initials: F
authors.lastname: Bourgaud
authors.forename: Eric
authors.initials: E
authors.lastname: Gontier
mesh_terms: D015139:Blotting, Southern; D003470:Culture Media; D005966:Glucuronidase / Q000235:genetics; D010957:Plasmids; D012038:Regeneration; D012231:Rhizobium / Q000235:genetics*; D032105:Ruta / Q000235:genetics* / Q000254:growth & development; D014170:Transformation, Genetic
chemical_list: D003470:Culture Media; D005966:Glucuronidase