| Different electroporation conditions were evaluated, toward the goal of transformation of Coffea arabica cv. Catimor. The tissues assayed were: embryogenic calli, leaf sections from in vitro plants, and somatic embryos in globular and torpedo stage obtained from cell suspensions. The effect of 1 or 24-hour pretreatment with an enzymatic solution (2% cellulase, 1% macerozyme) and electric field strength (375, 625, 875 V/cm) was evaluated. In all the experiments the tissues were incubated in ASP buffer (potassium aspartate) during three hours, and then one hour with plasmid DNA (pCambia3201, containing gus and bar genes) at room temperature. The electroporation was performed at a capacitance of 900 μF. The effect of the parameters evaluated was... |
