Transformación genética de citrange Troyer y de limonero Eureka con el gen reportero gf p mediada por Agrobacterium tumefaciens.

VELLICE, GABRIEL R.; VOJNOV, ADRIÁN; MARANO, MARÍA ROSA; CASTAGNARO, ATILIO P.

Boca Chica, República Dominicana

Otro; V Encuentro Latinoamericano y del Caribe de Biotecnología Agraria. REDBIO, 2004.; 2004

REDBIO

Genetic transformation were performed in citrange Troyer (Poncirus trifoliata (L) Raf. x Citrus sinensis (L) Osb.) and Eureka lemon (Citrus limon (L.) Burman). The EHA 105 supervirulent Agrobacterium tumefasciens strain carrying the binary vector pBIN19GFP was used. The binary plasmid contains the selectable marker nptII gene that confer kanamycin resistance and the gfp gene, Green Fluorescent Protein (GFP), as expression marker. The explants used in citrange Troyer were epicotyl sections from in vitro germinated seeds and mature stem section for lemon. After inoculation the explants were keep in dark at 25ºC during three days and then were subcultured to selection medium (MS basal medium, sucrose 3%, pH 5.7, kanamycin 100 mgL-1 and cefotaxime 500 mgL-1) added with 3 mgL-1 of Benzilaminopurine (BA) for citrange and 1 mgL-1 of BA for lemon. The 80% of inoculated Citrange Troyer formed shoots in selection medium after 1 month while 22% were positive for GFP detection. In the case of lemon, 70% of inoculated explants formed healthy callus in presence of kanamycin and 35% of explants were positive for GFP detection. The former mentioned callus showed no shoot regeneration after 3 months of culture.