Optimization of Agrobacterium-mediated transformation protocol of barley
To improve the efficiency of Agrobacterium-mediated transformation barley explants of three different types were involved in the work: callus tissue, mature and immature embryos. The vector contained the Fe-superoxide dismutase 1 (Fe-SOD1) gene as a target gene, which expression may contribute to the formation of non-specific plant resistance to stresses. Time of inoculation and co-cultivation of explants with Agrobacterium and effective methods of elimination of agrobacteria after co-cultivation stage was selected for barley callus tissue. Two fertile transgenic barley lines (cultivar Kupets) and one barley line (cultivar Belgorodsky 100) with Fe-SOD1 gene were obtained. The presence of alien insert was confirmed by PCR method. The obtained results can be used in the course of the Agrobacterium-mediated transformation of barley and other cereals.
Keywords: barley, transformation, Agrobacterium tumefaciens AGL0, superoxid dismutase, regeneration, molecular genetic analysis