1. Free Powerpoint TemplatesPage 1Free Powerpoint Templates Modification of cytokinin level in Hordeum vulgare Centre of the Region Haná for Biotechnological and Agricultural Research Mgr.Katarína Mrízová Mgr.Hana Pospíšilová, Ph.D

2. Free Powerpoint TemplatesPage 2 Constructs for barley transformation PromoterGeneTargeting β-GLU ZmCKX1secreted PHT ZmCKX1secreted RAF ZmCKX1secreted β-GLU AtCKX1vacuolar β-GLU AtCKX1cytosolic β-GLU AtCKX1secreted β-GLU AtCKX1plastidic

3. Free Powerpoint TemplatesPage 3 Root specific promoters Control Promoter of root abundant protein (RAF; hypotetical transcription factor) Promoter of phosphate transporter (PHT)

4. Free Powerpoint TemplatesPage 4 Root specific promoters Promoter of CK-specific β-glucosidase (β-GLU)

5. Free Powerpoint TemplatesPage 5 Results I. Constructβ-GLU::ZmCKX1PHT::ZmCKX1RAF::ZmCKX1 Transform. efficiency % 744 Relative expression level of transgene in roots 170-63311776646-6645 CKX activity roots up to 10-foldup to 2-fold Relative expression level of transgene in leaves 228561256-18051 CKX activity leaves up to 5-fold up to 122-foldup to 101-fold

6. Free Powerpoint TemplatesPage 6 PHT WT

7. Free Powerpoint TemplatesPage 7 WT RAF1 RAF2

8. Free Powerpoint TemplatesPage 8 WT β-GLU1 β-GLU2

9. Free Powerpoint TemplatesPage 9 PromoterGeneTargeting β-GLU ZmCKX1secreted PHT ZmCKX1secreted RAF ZmCKX1secreted β-GLU AtCKX1vacuolar β-GLU AtCKX1cytosolic β-GLU AtCKX1secreted β-GLU AtCKX1plastidic

10. Free Powerpoint TemplatesPage 10 Results II. gene AtCKX1 Transform. efficiency % Relative expression level of transgene in roots CKX activity roots Generation cytosolic7053-1000up to 10-foldT1 vacuolar18278-8664up to 7-foldT1 secreted1737-185up to 10-foldT1 plastidic7057-847up to 7-foldT1

11. Free Powerpoint TemplatesPage 11 T2 generation of transgenic barley with vacuolar AtCKX1 WT Homozygot WT Heterozygot

12. Free Powerpoint TemplatesPage 12 T2 generation of transgenic barley with vacuolar AtCKX1 WT Homozygot Heterozygot