Cloning and Bioluminescence Detection of Potato Genes

To obtain the coding sequence (CDS) of StBIN2, StCCJ9 (Soltu.DM.04G027660), and StSnRK2.2 (Soltu.DM.08G023690), we performed PCR amplification. The StBIN2 gene was cloned into the nLUC vector, and the StSnRK2.2 and StCCJ9 genes were inserted into the cLUC vector. Subsequently, the constructed plasmids were separately transformed into Agrobacterium tumefaciens GV3101. The transformed strains were cultured overnight in the YEB medium. Next, the StBIN2 bacterial solution was mixed with the StSnRK2.2 and StCCJ9 bacterial solutions, and the mixtures were infiltrated into tobacco leaves. After 36 h, the leaves were subjected to a photo recording using a molecular imaging system (Bio-Rad, Hercules, CA, USA). LUC enzyme activity detection was performed using a dual luciferase reporter assay kit (Vazyme, Nanjing, China) [18 (link)] for this experiment. The experiment was repeated three times biologically. The primers are listed in Supplemental Table S8.