Generation and Selection of Transgenic Olive Plants.

Olive ( L.) is one of the most important oil crops in the Mediterranean basin. Biotechnological improvement of this species is hampered by the recalcitrant nature of olive tissue to regenerate . In previous investigations, our group has developed a reliable mediated transformation protocol using olive somatic embryos as explants ( Torreblanca , 2010 ). Embryogenic cultures derived from radicles of matured zygotic embryos are infected with , AGL1 strain, containing a binary plasmid with the gene of interest and the selection gene. After a meticulous selection procedure, carried out using solid and liquid media supplemented with paromomycin, the putative transformed lines are established. A preliminary confirmation of their transgenic nature is carried out through PCR amplification. Afterwards, plants can be obtained through an efficient regeneration protocol, whose main characteristics are the use of a low-ionic-strength mineral formulation, a phase in liquid medium for synchronization of cultures and the use of semi-permeable cellulose acetate membranes for embryo maturation ( Cerezo , 2011 ). Final confirmation of transgene insertion is carried out through Southern or Northern analysis using leaf samples of regenerated plants.