Figure 5.

Generation of T-DNA knockout and overexpression lines. (A) Diagram of WRKY3 and WRKY4 and their T-DNA insertion mutants. (B) RNA gel blot analysis of wrky3 and wrky4 mutants. Wild-type and mutant plants were sprayed with SA (1 mM). The leaves were harvested 4 hours after treatment for total RNA isolation. After separation on the gels and blotting to nylon membranes, the blots were probed with corresponding gene-specific DNA fragments. (C) WRKY3 and WRKY4 expression in transgenic plants. RNA samples were prepared from leaves of 5-week-old wild-type (Col-0) and transgenic plants and probed with a WRKY3- or WRKY4-specific probe. Transgenic WRKY3 lines 3 and 8 and transgenic WRKY4 lines 7 and 13 contained a single T-DNA insertion in the genome and exhibited stable expression of their respective transgenes. Their F3 homozygous progeny plants were used in all the experiments in the study.