[Factors affecting Agrobacterium tumefaciens-mediated transformation of wheat (Triticum aestivum L.)]

Immature embryos and embryo-derived calli from two cultivars of winter wheat (Triticum aestivum L.), BAU146 and BAU170, were transformed with three strains of Agrobacterium tumefaciens, AGL-1, EHA105 and LBA4404 harboring expression vector p3301 or pBTAaB. Both vectors contained bar gene and p3301 contained also gus gene with an intron. The highest explant survival rate and transformation efficiency was obtained when the bacterial cell density was OD600 1.0 with 1 h of infection incubation. Higher osmotic treatment of the explants before inoculation had a positive effect on transformation, while addition of acetosyringone showed ambiguous one, depending on the explant types and bacterium strains. The efficiencies of transformation and transgenic plant regeneration were varied greatly with the bacterium strain, receptor genotype, explant type and its age and physiological state. After optimizing these factors, a large number of PPT-resistant calli and some of PPT-resistant plants were obtained. The resistant plantlet tested and 50% to 60% of the resistant calli were GUS-positive. The integration of foreign DNA into the genome of transgenic plants (3 out 6) was further confirmed by PCR and Southern Blot analysis.