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.
Key wordsAgrobacterium tumefaciens Ruta graveolens Hypocotyl Direct regeneration Transgenic plant Gus nptII
The authors would like to thank Andrew Saunders for help in improving the English quality of this manuscript.
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