Fig. 5

Molecular and biochemistry assays proving the transcriptional regulation of OsGAMYBL2 on GS3 gene. a Expression of GS3 gene in the STTM159 plants, GAMYBL2RNAi plants and ZH11 plants revealed by qRT–PCR (n = 3). The expression level in ZH11 was set as 1.0, and asterisks indicate significant differences compared with ZH11 plants as determined by Student’s t-test (**P < 0.01; *P < 0.05). b Sub-cellular localization of OsGAMYBL2 proteins in leaves of Nicotiana Benthamiana. Bar = 10 µm. c Luciferase strength of the transient coexpression of the effector and reporter constructs in tobacco leaves. d Measurement of the relative LUC/REN ratio after transient coexpression of the effector and reporter constructs in tobacco leaves. The values are means ± SDs (n = 3). The data were normalized to a value of 1 for the GAL4 DBD group. Asterisks indicate significant differences determined by Student’s t-test (**, P < 0.01). e EMSA assay of GAMYBL2 proteins and the promoter fragment of GS3 gene. f Image of Dual-LUC assay of GAMYBL2–GFP fused protein and GS3 gene promoter showing infloerescence strength. g The enzyme activity assay showing LUC/REN ratio of the experiments in E. The ratio in “Flag + pGS3” was set as 1.0, and asterisks indicate significant differences compared with “Flag + pGS3” as determined by Student’s t-test (**P < 0.01). Values are given as means ± SDs (n = 3)