Figure 8.

Binding activity of BnWRKY15 and BnWRKY33 for the three W‐box elements in Brassica napus. (a) Probes with either the first W‐box mutated or all three W‐box elements mutated were used in electrophoretic mobility shift assays. The W‐box element, TGAC core and mutated base are indicated in red. (b) The binding of the 33probe (WT) and Mt probe (with three W‐boxes mutated, as indicated in (a)) by BnWRKY15 and BnWRKY33 6× His fusion proteins. (c and f) The binding of the 33probe (WT) and mutated probes (sequences indicated in (a)) with the BnWRKY15 or BnWRKY33 6× His fusion proteins. (d and g) Relative luciferase activity from the transient expression analysis of P‐346 and different W1‐mutated promoter reporter plasmids cotransformed with 15SK or 33SK plasmids. The mutations of different W1‐mutated promoters correspond to the sequences of mutated probes in (a). A promoter containing four repeats of the first W‐box in place of the three W‐box regions is designated P‐4W1. The y‐axis represents different combinations of different effector plasmids and reporter plasmids. SK/0800 indicates a negative control (null effector plasmids and null reporter plasmids). The data represent the means ± standard errors (n ≥ 3). (e and h) The binding of three repeats of the first, second or third W‐box with BnWRKY15 or BnWRKY33 fusion proteins. Sequences of probes are as indicated in (a).