Fig. 8

DoDELLA1 yeast two-hybrid (Y2H) library screening assay. The N-terminus of the GRAS domain of DoDELLA1 with a binding function was used for Y2H screening. A Y2H screening and positive clone identification. Preliminary screening of 27 transformant colonies on SD medium without Trp and Leu. An additional 18 positive clones were obtained from the interaction between DoDELLA1 and other proteins in an in vitro X-α-gal activity assay, detected on SD medium without Trp, Leu, His and Ade. B Analysis of the interaction between DoDELLA1 and DoMYB39, DoDELLA1 and DoMYB308, and DoDELLA1 and DoWAT1, using a Y2H assay. All 18 positive clones were used for comparing sequences, and three key proteins (DoMYB39, DoMYB308 and DoWAT1) were used for Y2H rotary validation. An empty vector was used as the control. AD: pGADT7, a Y2H bait expression vector; BD: pGBKT7, a Y2H prey expression vector; empty, a negative control. SD/-Trp/-Leu: yeast culture medium without tryptophan and leucine; SD/-Trp/-Leu/-His/-Ade: yeast culture medium without tryptophan, leucine, histidine, and adenine; SD/-Trp/-Leu/-His/-Ade + X-α-Gal: yeast culture medium without tryptophan, leucine, histidine, and adenine, to which X-α-Gal was added