Fig. 6
OsMED16, OsWRKY45 and OsWRKY62 co-expression can further synergistically activate diterpenoid phytoalexin biosynthesis genes transcription. A Y1H experiment indicating OsWRKY62 binding to OsCYP99A3, OsKSL10 and OsDPF promoters. Negative control was performed with the empty vector pbait-ABA. Yeast cells were grown on SD-Leu and SD-Leu-Ura containing different concentrations of Aureobasidin A (AbA) at 30 °C for 3–5 d. 10–1, diluted 10 × 10–2, diluted 100 times. B Analysis of the effects of OsWRKY45 + OsMED16 and OsWRKY45 + OsMED16 + OsWRKY62 on proOsCYP99A3, proOsKSL10 and proOsDPF. Luminescence imaging was performed in leaves of N. benthamiana at 80 h after co-infiltration with the indicated vectors. C Schematic diagram of the trans-activation assay effector and reporter constructs. The reporter vector carries the renilla luciferase gene under the control of the 35S promoter and the firefly luciferase reporter gene under the control of the OsCYP99A3, OsKSL10, and OsDPF promoters. Ter, cauliflower mosaic virus terminator; REN, Renilla luciferase; LUC, firefly luciferase; 35sP, cauliflower mosaic virus 35S promoter. D Luciferase activity analysis using 35S: OsMED16-OsWRKY45 and 35S: OsMED16-OsWRKY45/35S: OsWRKY62 as effectors and ProOsCYP99A3:Luc, ProOsKSL10:Luc, and ProOsDPF: Luc as reporters, respectively. The LUC/REN activity ratios are the means ± SD; n = 3. Comparisons were performed with Student’s t-test. **P < 0.01