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. 2019 Mar 19;17(9):1770–1787. doi: 10.1111/pbi.13099

Figure 10.

Figure 10

PbrWRKY53 binds to and activates the promoters of PbrNCED1. (a) The prey and bait vectors used for yeast one‐hybrid assay. (b) Interaction of the PbrWRKY53 protein with labelled DNA probes for the cis‐elements of the PbrNCED1 promoter in the EMSA. (c) EMSA assays using affinity‐purified fusion protein His‐PbrWRKY53 incubated with biotin‐labelled probes of PbrNCED1 fragment containing wild type (TTGACC) or mutated W‐box element (TCACGT); the nonlabelled fragment was used as a competitor; −, absence; +, presence. (d) Transient expression assay of the promoter activity using tobacco protoplasts co‐transformed with the effector and each of the reporters containing the normal W‐box element (P2) or mP2 containing mW‐box element. (e) Schematic structures of the effector (p1301‐PbrWRKY53) and reporter vector (W‐box‐m35S‐GUS or m35S‐GUS) used for transient expression analysis. (f) GUS staining of representative leaf pieces infiltrated with only reporters (top panel) or those coinfiltrated with the effector and the reporters (W‐box‐m35S‐GUS; bottom panel). − indicates the absence of effector (top panel) or reporters (bottom panel).