Yeast one-hybrid (Y1H) assay

The assay of Y1H was carried out on the basis of the Matchmaker Gold Yeast One Hybrid System protocol (Clontech, CA, USA). To construct a prey vector, the CDS of SlNAP1 was amplified and merged into pGADT7. The promoter sequences of SlGA2ox1 and SlGA2ox5, covering 1500 bp, were obtained from NCBI databases (https://www.ncbi.nlm.nih.gov/) and merged into pAbAi to form a bait vector. The linearized pAbAi-proSlGA2ox1 and pAbAi-proSlGA2ox5 plasmids were transformed into Y1H Gold yeast strains. Screening for minimal inhibitory concentration of Aureobasidin A (AbA) was performed to avoid any self-activation instances. The bait yeast strains were transformed with the pGADT7-SlNAP1 vector and cultured on SD medium lacking Leu (SD/-Leu) at 30 °C for 2–3 days, both with and without AbA. A control was established using the pAbAi and pGADT7 plasmids. These experiments were repeated three times, yielding similar results, and a representative image is provided.