We developed an Agrobacterium tumefaciens-mediated transformation system for Eucalyptus urophylla using hypocotyl explants. Antibiotic concentrations, pre-culture times, pH of the inoculation medium and co-culture times were optimised. Pre-cultured hypocotyl explants were co-cultured with A. tumefaciens strain EHA105 harbouring the binary vector pPBR-2 containing the Rs-AFP2 gene, which encodes an antifungal protein, under the control of the prp1-1 promoter, for six days and were then transferred to selective callogenesis-inducing medium containing kanamycin and cefotaxime. Calluses developed ...