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Agrobacterium-Mediated Transformation of Ruta graveolens L

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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 C58C1RifR containing plasmid pTDE4 harbouring neomycin phosphotransferase (npt II, kanamycin resistance) and β-glucuronidase encoding genes. The Southern blot analysis has shown that 78% kanamycin resistant plants contain gene encoding β-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.
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