FIGURE 6.

TALEs directly induce expression of target rice promoters in reporter studies in N. benthamiana. (A,B) 1,000 bp upstream of the ATGs of the TALE target genes were amplified from rice cultivar Nipponbare DNA and cloned in front of a promoterless uidA reporter gene. Artificial TALEs were assembled with RVD sequences shown in Figure 5 and Hax3 N- and C-terminal regions under control of a 35S promoter. A. tumefaciens strains delivering the reporter constructs and strains delivering the TALE expression constructs were co-inoculated into N. benthamiana leaves and β-glucuronidase measurements were performed 2 dpi. Quantitative GUS activity measurements were performed three times with samples obtained as described above. Error bars represent standard deviation between triplicates. The statistical significance between samples with and without corresponding TALEs is indicated by p-values resulting from an unpaired t-test and fold changes are marked in blue. The TALE Hax3 is used as a negative control in samples labeled without corresponding TALE. Histochemical GUS staining of leaf disks and quantitative GUS activity measurements were done in parallel from the same plants. Leaf disks were stained in GUS staining solution and destained in 96% ethanol. One representative leaf disk is shown. (C) Schematic overview of experimental setup.