Figure 4

Transactivation of OsNAC4 and WRKY62 genes by WRKY45. (a) Transactivation of WRKY62 and OsNAC4 genes in transient trans-activation assays. Upstream and intragenic region of WRKY62 (-1,070 – +1,838) or OsNAC4 (-1,000 – +1,272) fused to the hrLUC gene in pGL4.7 vector (Promega) were used as reporter genes. Firefly LUC gene driven by the maize ubiquitin promoter was used as a reference gene. Reporter genes were co-delivered into rice coleoptiles with the effector genes (120 ng) with (+) or without (-) WRKY45 cDNA, and the reference gene (firefly LUC). Luciferase activities were determined in three biological replicates. Average values are shown with SD. (b) Mutation analysis of W-boxes in OsNAC4 upstream region in transient trans-activation assays. Duplicates of the OsNAC4 upstream sequence (-504–-485) containing wild-type (wNAC4) or mutant (mNAC4) forms of W-box-like sequences fused to firefly LUC gene were used as reporter genes. Reporter genes were co-delivered with 500 ng of effector genes with (+) or without (-) WRKY45 cDNA described above, together with the hrLUC reference gene. Luciferase activities were determined in three replicates. Average values are shown with SD.