2.7 Yeast one-hybrid assay (Y1H)
Yeast one-hybrid assay (Y1H) was performed to verify the DNA-protein interaction following the manual of the Matchmaker one-hybrid system (Clontech). The full-length cDNA of OsLHY was cloned to fuse with the GAL4 activation domain in the pGAD7-Rec2 vector. The promoters of flowering related genes were amplified to insert into the pHIS2 vector to generate reporter constructs. Subsequently, these reporter plasmids were separately co-transformed with the fusion plasmid pGADT7-OsLHY into the yeast Y187. The co-transformed yeast cells were grown on the SD/-Leu/-Trp/-His and SD/-Leu/-Trp/-His media containing 50 mM 3-amino-1,2,4-triazole (3-AT) in a 30℃incubator for 3-5 d. Relative primers are shown in Supplemental Table 1.