Establishment of Agrobacterium-mediated genetic transformation system in Dahlia

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  • Establishment of <i>Agrobacterium</i>-mediated genetic transformation system in <i>Dahlia</i>

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An efficient system for Agrobacterium-mediated transformation was established in dahlia (Dahlia×pinnata) ‘Yamatohime’. Mass of shoot primordia (MSP) induced on MS medium supplemented with 10 mg l−1 TDZ were inoculated with Agrobacterium tumefaciens strain EHA101 (pIG121-Hm) harboring both β-glucuronidase (GUS) and hygromycin resistant genes. After 2 days of co-cultivation, the MSP were transferred to a selection medium containing hygromycin with meropenem for bacterial elimination. Shoots were successfully regenerated from survived MSP on hormone-free medium without hygromycin and they rooted on hormone-free medium containing hygromycin. The hygromycin-resistant plants thus obtained showed histochemical blue staining for GUS. Transformation of plants was confirmed by PCR and Southern blot analyses. Transgenic ‘Kokucho’ were also produced by using the same transformation procedure, suggesting wide applicability of this Agrobacterium-mediated transformation procedure for other dahlia cultivars.

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