FIGURE 5.

Interaction test of PP2Cs with PYLs and SnRK2s in the yeast two-hybrid assay. (A) AtPP2Cs fused to the GAL4 Activation Domain (AD) were co-transformed in yeast with CaPYLs cloned in frame with the GAL4 binding domain (BD) of pGBKT7, with combinations shown in figure. (B) CaHAI fused to the AD in pGADT7 was expressed in yeast with AtPYLs in pBDGAL4, the different combinations are shown. (C) CaPP2Cs were fused to the AD in pGADT7 and co-transformed with CaPYLs cloned in pGBKT7. (D) CaPP2Cs in pGADT7 were co-transformed in combination with AtSnRK2s cloned in pGBKT7. (E) AtPP2Cs in pACT2 were co-transformed in yeast with CaSnRK2s in pGBKT7, with combinations shown in figure. (F) Co-transformants of yeast containing CaPP2Cs fused to the AD in pGADT7 in combination with CaSnRK2s in pGBKT7. Co-transformations of the pepper PYL/SnRK2/PP2C orthologs with appropriate complementary empty vectors are shown as negative controls. Yeast cells grown on synthetic media (–W/–L) and on synthetic, selective media without (–W/–L/–H/–A) or, where indicated, with 50 μM ABA (–W/–L/–H/–A+ABA) are shown. Pictures were taken after 3 days of incubation at 30°C.