Genome wide analysis and functional identification of MdKCS genes in apple

Apple fruit is covered by cuticle wax, which plays important roles protecting fruits from adverse environmental conditions. β-Ketoacyl-CoA synthase (KCS) is the key rate-limiting enzyme in plant wax synthesis. In this study, we identified 28 KCS gene family members from apple (Malus × domestica Borkh.) by homology analysis. Multi-sequence alignment and phylogenetic analyses revealed that the 28 MdKCS genes were divided into four subgroups, including KCS1-like, FAE1-like, FDH-like, and CER6. A chromosomal localization analysis revealed that 27 apple KCS genes were located on 11 chromosomes, while MdKCS28 was localized to the unassembled genomic scaffold. Most of the MdKCS proteins were hydrophilic proteins and they had similar secondary and tertiary structures. The prediction of cis-acting elements of the MdKCS gene promoters suggested that the MdKCS genes may be widely involved in hormone signaling and the stress response. Furthermore, the quantitative real-time polymerase chain reaction results showed that eight MdKCS genes were highly expressed in the apple pericarp, and were significantly induced by drought, abscisic acid (ABA), and NaCl treatments. We transformed the MdKCS21 gene into apple calli, and found the MdKCS21 overexpressing transgenic apple calli exhibited higher tolerance to ABA treatment. Finally, the MdKCS proteins were localized to the endoplasmic reticulum and vacuolar membrane by confocal laser microscopy. This study established a foundation to further analyze the function of KCS genes and provided candidate genes for molecular improvement of wax content in apple.