ISHS Acta Horticulturae 577: VII International Symposium on Plum and Prune Genetics, Breeding and Pomology
AGROBACTERIUM - MEDIATED TRANSFORMATION OF PLUM (PRUNUS DOMESTICA L.) |
| Authors: | S.D. Yancheva, PH. Druart, B. Watillon |
| Keywords: | plum, transformation, regeneration, leaves, NPT II gene, GFP gene |
| Abstract:
An efficient adventitious shoot regeneration protocol from leaf explants of the Bulgarian local cultivar ‘Kyustendilska sinya’ and the Belgian – ‘Altesse simple’ (Quetsche) clone was developed. Sufficient regeneration frequency was achieved on MS basal medium supplemented with 1.65 mg/l Thidiazuron (TDZ) and 0.5 mg/l Indole-3-butyric acid (IBA), 2% sorbitol and 1% sucrose under red fluorescent light. Putative transgenic plants were obtained from wounded leaves co-cultivated with EHA 105 and LBA4404 Agrobacterium tumefaciens strains, containing a pBin19 plasmid derivative carrying neomycin phosphotransferase (NPT II) and green fluorescent protein (GFP) genes under the control of CaMV 35S promoter. Only three putative transgenic plans were selected on a medium with 25 and 50 mg/L kanamycin. The integration of the NPT II marker gene was detected by PCR in leaves of the regenerated plants. | |
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