Figure 2: Outlines of the rice transformation protocol : Embryogenic nodular units (B: arrows) released from the primary, embryo scutellum-derived callus (A) of the japonica cultivar Nipponbare are transferred to fresh medium to reach an optimal size (C) and immersed into liquid co-culture medium containing EHA105 or LBA4404 cells at an OD600 of 1, for 15min, then blot dried and transferred to Petri dishes containing solid, co-culture medium, for a 3 day incubation period in the dark at 25°C. The procedure for selecting hygromycin-resistant cell lines (D : here visualized through GFP activity at the surface of a co-cultured callus, 14 days after transfer to selective medium) using 2-3 weeks subcultures on different selection media (E and F: 28 and 35 days after the first transfer of co-cultured calli to selective medium) and evolving transgenic rice plants (G,H,I) is detailled in Sallaud et al, 2003, Theor Appl Genet, 106: 1396-1408)