Systematic analysis reveals O-methyltransferase gene family members involved in flavonoid biosynthesis in grape

Abstract

O-methyltransferases (OMTs) are an important group of enzymes involved in the methylation of various secondary metabolites, including flavonoids. However, the features and functions of OMTs have not been comprehensively studied in grape (Vitis vinifera), a rich source of methylated flavonoids. Here, 47 OMT members were identified in grape genome. They were unevenly distributed on grape chromosomes and some genes were tandem duplicated, indicating the role of duplication processes in the expansion of this gene family. Based on the phylogenetic relationship, these OMTs were clustered into CCoAOMT and COMT subclades, which were further supported by the results of conserved motif and gene structure analysis. Correlation analysis revealed that three members (VvCCoAOMT1, VvCCoAOMT4, and VvCOMT1) were potentially involved in the synthesis of most methylated flavonoids in the berry skins. Expression profiling based on RNA-seq data and qRT-PCR experiments indicated that VvCCoAOMT1 and VvCCoAOMT4 had specific and high expression in berry skins, and responded to abscisic acid and high temperature treatments; and that VvCOMT1 expression was significantly induced during berry development and UVC treatment. Cis-regulatory element analysis suggested important roles of OMTs in growth, development, and defense against stresses. We further demonstrated the transcriptional regulation of VvCCoAOMT4 by VvMYBA1, a master regulator of grape berry anthocyanin, and verified the protein localization of VvCCoAOMT4 in membrane and nucleus. These findings facilitate a better understanding of the characteristics of OMT gene family, especially of the potential members involved in the formation of O-methylated flavonoids in grape.

Keywords: Anthocyanin; Flavonoid; Grape (Vitis vinifera); O-methyltransferase.