Fig. 4

PbrERF114 directly binds to the PbrACS3 promoter. A Predicted GCC-box and DRE elements were identified in brACS1, PbrACS3, and PbrACO1 promoters. B PbrERF114 induced LUC expression controlled by PbrACS1, PbrACS3, and PbrACO1 promoters. Standard errors and analysis of variance were determined using Student’s t-test. Single and double asterisks represent significant differences at P < 0.05 and 0.01, respectively. C The Y1H assay evaluated the interaction between PbrERF114 and the predicted GCC-box elements in the PbrACS1 promoter. D EMSA examined the binding of PbrERF114 to the PbrACS1 promoter. + and − denote the presence and absence of recombinant PbrERF114-HIS, biotin-labeled probe, cold probe, and mutant cold probe, respectively. The cold probe concentrations were tenfold (10 ×), 50-fold (50 ×), and 200-fold (200 ×) those of the labeled probes