Fig. 4

Binding of SlCOL1 to the SFT promoter. A Schematic diagram of the 2533-bp SFT promoter region. Seven cis-elements were identified in the promoter of SFT. TSS, transcription start site. B Yeast-one hybrid (Y1H) analysis of SlCOL1 binding to the different core sequences of the SFT promoter. Five constructs containing five different promoter fragments (SFT1 to SFT5) were used in Y1H assays. The bait vectors, SFT1 to SFT5, and the SlCOL1-containing prey vector were introduced into the yeast strain Y1H Gold. The enhanced resistance to antibiotic aureobasidin A (AbA) indicated an interaction between the bait and prey. Co-transformation of the bait vectors, SFT1 to SFT5, with either pGADT7 or pGADT-Rec2–53 served as negative and positive controls, respectively. C GAL4/UAS-based analysis on SlCOL1 binding to the SFT promoter. The promoter of SFT was fused to an open reading frame encoding the GUS protein (ProSFT-GUS). SlCOL1 was expressed from the pHELLSGATE8 vector (35S-SlCOL1). The resulting constructs were transiently co-expressed in the leaves of N. benthamiana. ProSFT-GUS and the empty vector pHELLSGATE8 were included as controls. Values are presented as means ± SE (n = 3). The asterisks indicate statistically significant differences. **, P < 0.01. nd, Not detected