Fig. 10: Direct repression of MdRAV1 on MdABI promoters.

a EMSA of MdRAV1 binding to the MdABI3 (MD10G1169900) promoter in vitro. Each biotin-labeled DNA probe was incubated with MdRAV1-His protein. Mutation probes of P1 had a mutated W-box (CAACA was replaced with TGGGG). b EMSA of MdRAV1 binding to the MdABI4 (MD07G1224400) promoter in vitro. Each biotin-labeled DNA probe was incubated with MdRAV1-His protein. Mutation probes of P1 had a mutated element (CAACA was replaced with TGGGG). c Interaction of MdRAV1 with the promoter fragment of MdABI3 and MdABI4 in the yeast one-hybrid assay. The constructs were cotransformed into yeast Y187. The yeast was cultured on a culture medium devoid of tryptophan and histidine (SD/-T/-H) and then screened for lack of tryptophan, leucine, and histidine (SD/-T/-L/-H) with 20 and 80 mM of 3-amino-1,2,4-triazole (3-AT). d, e GUS activity in transgenic calli as indicated