Agrobacterium-mediated stable transformation of cell suspension cultures of barley (Hordeum vulgare)

摘要: Barley (Hordeum vulgare L. cvs. Igri and Dissa) cell suspension cultures, which had been initiated from immature embryo-derived (IED) microspore-derived (MSD) callus, were co-cultivated with various Agrobacterium tumefaciens strains. The T-DNA vectors contained visually-detectable marker genes (C1/Lc orgusA-intron), as reporters of transient transfer, also drug resistance (hph or bar) to facilitate selection stably transformed lines. A set normal binary in a super-virulent strain [EHA101(pBECKS)] super-binary vector [LBA4404(pTOK233)] used this study. Cells the cultures received able proliferate under regimes number hygromycin- phosphinothricin-resistant barley callus lines isolated expressed co-transferred gusA gene. To ensure homogeneity lines, prolonged tissue culture but these resulted loss capacity regenerate plants transgenic frequency recovery ranged 0.3% 2.9%. Southern blot analyses confirmed presence demonstrated them be associated DNA was distinct that original plasmid. Furthermore, independent showed diverse patterns hybridising bands. These data suggest fragment maintained through integration into genomes