Figure 2.

Functional Analysis of the R2R3 MYB Proteins from WT and Aft Plants.

(A) Transactivation of the SlDFR promoter driving firefly luciferase in protoplasts with effector plasmids containing the MYB SlAN2, SlANT1, SlANT1like, or SlAN2like genomic sequences from WT or Aft plants, in combination with the effector plasmid containing the bHLH factor SlAN1. Data are expressed as relative luciferase activity (RLU) (FireflyLuc/RenillaLuc) with the value of the promoter basal level set to 1 and are means of four biological replicates ± SE. One-way ANOVA with Tukey's HSD post hoc test was performed. ns, P > 0.5; ****P ≤ 0.0001.

(B) Transactivation of the SlDFR promoter driving firefly luciferase in protoplasts with effector plasmids containing the MYB SlAN2like genomic sequence from WT or Aft plants, in combination with effector plasmids containing the bHLH factor SlAN1 or SlJAF13. Data are expressed as RLU with the value of the promoter basal level set to 1 and are means of four biological replicates ± SE. One-way ANOVA with Tukey's HSD post hoc test was performed. Different letters indicate significant differences at P ≤ 0.05.

(C) Anthocyanin accumulation in tobacco leaves agroinfiltrated with effector plasmids containing the SlAN2like genomic sequence cloned in WT or Aft plants expressed with or without the effector plasmid containing the bHLH factor SlAN1. White dotted circles indicate the agroinfiltrated areas.

(D) Quantification of the anthocyanins produced in the areas of tobacco leaves agroinfiltrated with WT or Aft SlAN2like in combination with SlAN1. Anthocyanins are expressed in μg petunidin-3-(p-coumaroyl rutinoside)-5-glucoside g⁻¹ fresh weight (FW). Data are means of eight biological replicates ± SE. One-way ANOVA with Tukey's HSD post hoc test was performed. Different letters indicate significant differences at P ≤ 0.05.

(E)Solanum chilense mature fruits (picture reproduced with the permission of the author from https://giorgetta.ch/fl_solanaceae_solanum_chilense.htm).

(F) Transactivation of the SlDFR promoter driving the firefly luciferase gene in protoplasts transfected with effector plasmids containing SlAN2like genomic sequences from WT or Aft plants or ScAN2like genomic sequence from S. chilense, in combination with the effector plasmid containing the bHLH factor SlAN1. Data are expressed as RLU with the value of the promoter basal level set to 1 and are means of four biological replicates ± SE. One-way ANOVA with Tukey's HSD post hoc test was performed. Different letters indicate significant differences at P ≤ 0.05.