Figure 3

Analyses of the bindings of ThbZIP1 to C-, G- and A-box. A: (a-d): Analysis of the bindings of ThbZIP1 to C-, G-, A-box and their mutated sequences by yeast one-hybrid analysis. The yeast cells were grown on different intensities of selective dropout medias: SD/- Trp-Leu/-His (TDO) + 3-AT (3-AT concentrations, a: 30 mM, b: 40 mM, c: 50 mM, d: 60 mM). B: A diagram of the reporter and effector vectors. C: The coexpression of reporter and effector vector in tobacco leaves. pCAM-C, -G -A, -CM2: three tandem of C-, G-, A-box, or their mutant CM2 was respectively fused to a minimal promoter (-46 to +1) to drive GUS. pROKII-ThbZIP1: the ORF of ThbZIP1 was under the control of CaMV 35S promoter. D: GUS activity assay of the coexpression of reporter and effector plasmid. CaMV35S: The transformation of pCAMBIA1301 alone (positive control). The transformation of the reporter plasmids alone were used as negative controls. All assays were repeated three times and error bars indicate SE.