Clontech one hybrid system

The Clontech™ one‐Hybrid System (Clontech, Dalian, China) was used in this study. The CDS of potential transactivators were fused with GAL4 AD domain in pB42AD (Clontech, Dalian, China), and the promoter region of Wx were cloned into pLacZ2u (Clontech, Dalian, China). Yeast strain EGY48 was transformed with indicated plasmids and grew on SD/‐Ura/‐Trp plates, and then strike on SD/‐Ura/‐Trp plates containing 2% glactose, 1% raffinose, 1 × BU salts and 80 mg/L X‐Gal (Clontech, Dalian, China). The interaction was confirmed by the visualization of blue colonies on the medium. NF‐YB1‐pB42AD and SUT4‐pLacZ2u were used as CK+ (Bai et al., 2015). The empty vectors pLacZ2u and pB42AD were used as negative control.

In this study, the Clontech™ one-hybrid system (Clontech, Dalian, China) was used. The CDS of the potential trans-activator was fused to the GAL4 AD structural domain in the pB42AD vector (Clontech, Dalian, China), and the promoter sequences of Wx, SLRL2, and bHLH144 genes were cloned into pLacZi vector (Clontech, Dalian, China), respectively. Yeast strain EGY48 was transformed and cultivated on the corresponding SD dropout medium for selection (Clontech, Dalian, China). Protein-DNA interactions were confirmed by the detection of blue colonies on the selection medium.