2.12 LUC reporter assay
The LUC reporter assay was conducted following a previously described protocol (An et al. , 2017). The MdNCED1 and MdNCED3promoters were amplified, and then cloned into the pGreenII 0800-LUC vector resulting in the reporter constructs MdNCED1 pro:LUC andMdNCED3 pro:LUC. The MdWRKY33 ORF was cloned into the pGreenII 62-SK vector to generate the effector (35Spro:MdWRKY33 ). The LUC reporter assay was conducted using the Dual-Luciferase® Reporter Gene Assay Kit from Yeasen Biotechnology (Shanghai, China), referring to the manufacturer’s instructions.
RESULTS