Transformation of barley scutellum protoplasts: regeneration of fertile transgenic plants

A system for barley transformation via polyethyleneglycol-mediated DNA upta ke into protoplasts isolated directly from scutella and the regeneration of transgenic plants is reported. Scutellum protoplasts (cv. Clipper, an Aust ralian malting cultivar) were co-transformed with plasmids Act 1-DGUS, cont aining the marker uidA gene, and pCaIneo, which contains the selectable mar ker neomycin phosphotransferase gene. Protoplast-derived calluses were sele cted on medium containing the antibiotic G418 (25 and 15 mg.1(-1)) and macr oscopic antibiotic resistant colonies were recovered. Fertile plants were r egenerated from a callus line and molecular analysis confirmed transgene in tegration.