Fig. 4
OsMED16 and OsWRKY45 synergistically activate diterpenoid phytoalexin biosynthesis genes transcription. A Y1H experiment indicating binding of OsWRKY45 to the promoters of OsCYP99A3, OsKSL10 and OsDPF. 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 W box position diagram of OsCYP99A3, OsKSL10 and OsDPF promoters. 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. 35sP, cauliflower mosaic virus 35S promoter; REN, Renilla luciferase; Ter, cauliflower mosaic virus terminator; LUC, firefly luciferase. D Analysis of the effects of OsWRKY45 and OsMED16 on proOsCYP99A3, proOsKSL10 and proOsDPF. Luminescence imaging was performed in leaves of N. benthamiana at 80 h after co-infiltration with the indicated vectors. E Luciferase activity analysis using 35S: OsWRKY45 and 35S: OsMED16-OsWRKY45 as effectors and proOsCYP99A3:Luc, proOsKSL10:Luc, and proOsDPF: Luc as reporters, respectively. Values of LUC/REN activity ratio are mean ± SD; n = 3. The Student's t-test was used for comparison. *P < 0.05; **P < 0.01