23 research outputs found
Sour orange bud regeneration and in vitro plant development related to culture medium composition and explant type
Cultivo inicial in vitro de gemas axilares de Ananas comosus (L.) Merr., em meio líquido/sólido, na presença/ausência de luz
Eficiência de transformação genética de citrange 'carrizo' com duas construções gênicas
Growth regulators, culture media and antibiotics in the in vitro shoot regeneration from mature tissue of citrus cultivars
Genetic transformation of sweet oranges with the D4E1 gene driven by the AtPP2 promoter
Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange.
The development and optimization of efficient transformation protocols is essential in new citrus breeding programs, not only for rootstock, but also for scion improvement. Transgenic 'Hamlin' sweet orange (Citrus sinensis (L.) Osbeck) plants were obtained by Agrobacterium tumefaciens-mediated transformation of epicotyl segments collected from seedlings germinated in vitro. Factors influencing genetic transformation efficiency were evaluated including seedling incubation conditions, time of inoculation with Agrobacterium and co-culture conditions. Epicotyl segments were adequate explants for transformation, regenerating plants by direct organogenesis. Higher percentage of transformation was obtained with explants collected from seedlings germinated in darkness, transferred to 16 hours photoperiod for 2-3 weeks, and inoculated with Agrobacterium for 15-45 min. The best co-culture condition was the incubation of the explants in darkness, for three days in culture medium supplemented with 100 mM of acetosyringone. Genetic transformation was confirmed by performing ß-glucoronidase (GUS) assays and, subsequently, by PCR amplification for the nptII and GUS genes.200