Figure 4.

MYBS1 and MYBGA Interact Physically and Coactivate the α-Amylase Promoter.

(A) BiFC assay. Barley aleurones were cotransfected with Ubi:nYFP-OsMYBS1 and Ubi:cYFP-OsMYBGA and incubated in the presence of Glc for 24 h. YFP was detected by confocal microscopy. See also Supplemental Figure 7 online.

(B) GAL4-UAS two-hybrid assay. Rice aleurones were cotransfected with effectors Ubi:GBD-OsMYBS1 and Ubi:GAD-OsMYBGA and reporter (5xUAS)-CaMV35S:Luc.

(C) MYB-αAmy8 two-hybrid assay. Rice aleurones were cotransfected with effectors Ubi:OsMYBS1 and Ubi:OsMYBGA and reporter 2xαAmy8 GARC-CaMV35S:Luc.

In (B) and (C), transfected rice aleurones were incubated in the presence of Glc for 24 h, and luciferase activity was assayed. The luciferase activity in rice aleurones bombarded with effectors Ubi:GAD and Ubi:GBD in (A) or reporter 2xαAmy8 GARC:Luc in (B) was set to 1×, and other values were calculated relative to this value.

[See online article for color version of this figure.]