Transformation of somatic embryos of mango by biobalistics.
Chavarri M., Vegas A., Zambrano A. Y., Demey J. R.
Author Affiliation: Universidad Central de Venezuela (UCV), Venezuela.
Interciencia 29 : 261-266
Abstract : Somatic embryos of mango cultivars Haden, Madame Francis and Kent were bombarded. The maximum dose of the herbicide phosphinothricin [glufosinate] (PPT) that somatic embryos survived was determined. Embryos were exposed in Gamborg, Miller and Ojima media containing 0, 0.5, 1 and 2 mg PPT/litre during 3 months and subculturing every month to evaluate fresh and dry weight. The embryos were bombarded with the plasmid CAMBIA 3201 with GUS and BAR genes, using 2 tungsten particle sizes (0.7 and 1.3 µm), pressure of 80 psi, bombardment distances of 10 and 16.5 cm, and 5 µg of DNA. Embryos were selected using 0.5 mg PPT/litre as selective agent. Particle size of 0.7 µm and the bombardment distance of 16.5 cm were selected. After 3 months, clones of cv. Kent survived all concentrations, while non-transformed embryos survived only at 0.5 mg/litre. Haden and Madame Francis cultivars were sensitive to the bombardment and selection conditions, and did not survive the process. Embryo survival was 4%, and 3 clones per 0.5 g of tissues bombarded were regenerated. Enzymatic activity of transient expression of the GUS gene was observed and the incorporation of GUS and BAR genes was demonstrated by means of PCR.