Genome-wide identification of the CER1 gene family in apple and response of MdCER1-1 to drought stress

Plant cuticular wax was a major consideration affecting the growth and quality of plants through protecting the plant from drought and other diseases. According to existing studies, CER1, as the core enzyme encoding the synthesis of alkanes, the main component of wax, can directly affect the response of plants to stress. However, there were few studies on the related functions of CER1 in apple. In this study, three MdCER1 genes in Malus domestica were identified and named MdCER1-1, MdCER1-2, and MdCER1-3 according to their distribution on chromosomes. Then, their physicochemical properties, sequence characteristics, and expression patterns were analyzed. MdCER1-1, with the highest expression level among the three members, was screened for cloning and functional verification. Real-time fluorescence quantitative PCR (qRT-PCR) analysis also showed that drought stress could increase the expression level of MdCER1-1. The experiment of water loss showed that overexpression of MdCER1-1 could effectively prevent water loss in apple calli, and the effect was more significant under drought stress. Meanwhile, MdYPB5, MdCER3, and MdKCS1 were significantly up-regulated, which would be bound up with waxy metabolism. Gas chromatography-mass spectrometer assay of wax fraction makes known that overexpression of MdCER1-1 apparently scaled up capacity of alkanes. The enzyme activities (SOD, POD) of overexpressed apple calli increased significantly, while the contents of proline increased compared with wild-type calli. In conclusion, MdCER1-1 can resist drought stress by reducing water loss in apple calli epidermis, increasing alkanes component content, stimulating the expression of waxy related genes (MdYPB5, MdCER3, and MdKCS1), and increasing antioxidant enzyme activity, which also provided a theoretical basis for exploring the role of waxy in other stresses.