UDP-glucosyltransferase PpUGT85A2 controls volatile glycosylation in peach

The monoterpene linalool is a major contributor to aroma and flavor in peach (Prunus persica) fruit. It accumulates during fruit ripening, where up to ~40% of the compound is present in a non-volatile glycosylated form, which affects flavor quality and consumer perception by retronasal perception during tasting. Despite the importance of this sequestration to flavor, the UDP-glycosyltransferase (UGT) responsible for linalool glycosylation has not been identified in peach. UGT gene expression during peach fruit ripening and among different peach cultivars was analyzed using RNA sequencing, and transcripts correlated with linalyl-β-d-glucoside were selected as candidates for functional analysis. Kinetic resolution of a racemic mixture of R,S-linalool was shown for PpUGT85A2, with a slight preference for S-(+)-linalool. PpUGT85A2 was shown to catalyze synthesis of linalyl-β-d-glucoside in vitro, although it did not exhibit the highest enzyme activity between tested substrates. Subcellular localization of PpUGT85A2 in the cytoplasm and nucleus was detected. Application of linalool to peach leaf disks promoted PpUGT85A2 expression and linalyl-β-d-glucoside generation. Transient expression in peach fruit and stable overexpression in tobacco and Arabidopsis resulted in significant accumulation of linalyl-β-d-glucoside in vivo. Taken together, the results indicate that PpUGT85A2 expression is a major control point predicting linalyl-β-d-glucoside content.