Agrobacterium-mediated transformation of Ruta graveolens L.
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.
Lièvre
Karine
K
UMR Nancy-Université-INRA Agronomie et Environnement Nancy-Colmar, Vandoeuvre-Lès-Nancy, France.
Tran
Thi Lê Minh
TL
Doerper
Sébastien
S
Hehn
Alain
A
Lacoste
Paul
P
Thomasset
Brigitte
B
Bourgaud
Frédéric
F
Gontier
Eric
E
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
D003470:Culture Media; D005966:Glucuronidase