Agrobacterium tumefaciens-mediated transformation of blackberry (Rubus fruticosus L.)

Abstract

The aim of work was to elaborate the system for Agrobacterium tumefaciens-mediated transformation and transgenic plant regeneration in Rubus fruticosus L. using explants from mature plants. Regeneration of transgenic shoots was achieved from cut ends of petioles completely immersed in the medium using flag explants cultivated vertically on MS medium with 1 mg l−1 TDZ and 0.02 mg l−1 IBA followed by transfer on medium with 1 mg l−1 BAP, 0.02 mg l−1 IBA and 0.1 mg l−1 GA3, both supplemented with 10 and 15 mg l−1 hygromycin after transformation by A. tumefaciens strain LBA 4404/pCambia 1304. Four putative transgenic plants of cv. ‘Cacanska Bestrna’ were rooted, acclimatized and analysed. Transgenic character of the tissue from analysed plants was confirmed by nested PCR and subsequent sequencing.