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AGROBACTERIUM - MEDIATED TRANSFORMATION OF PLUM (PRUNUS DOMESTICA L.)
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2002
Year
Bulgarian Local CultivarEngineeringBotanyGeneticsGreen Fluorescent ProteinBiosynthesisHorticultural PlantHorticultural SciencePlant BiologyPlant-microbe InteractionAgricultural BiotechnologyBiologyBiotechnologySynthetic BiologyGenetic EngineeringPlant Cell CultureSynthetic Plant BiologyLeaf ExplantsMicrobiologyPrunus Domestica LMedicinePlant Physiology
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.