Transgenic production using particle bombardment in Egyptian barley

ABSTRACT

A system for Egyptian barley [Hordeum vulgare L. cv Giza 123] transformation via biolistic bombardment has been established. Immature embryos 1.0-1.5 mm in length were aseptically isolated from barley plants cultivated in the open field. Embryos were then cultured on callus induction medium with scutellum side up. Scutellum derived calli were bombarded with the plasmid pAHC25, containing the marker gene uidA as a reporter gene and bar gene as a selectable marker gene. Bombarded tissues were transferred into callus induction medium containing bialaphos for selection. Calli grown on selection medium were transferred into regeneration medium supplemented with bialaphos. Putative transgenic callus events produced putative transgenic plants. Integration of transgenes was confirmed by molecular analysis. The expression of uidA genes was studied by histochemical staining while the expression of bar gene was studied by leaf paining assay technique