Monoterpenols serve various biological functions and accumulate in grape (Vitis vinifera), where a major fraction occurs as nonvolatile glycosides. We have screened the grape genome for sequences with similarity to terpene URIDINE DIPHOSPHATE GLYCOSYLTRANSFERASES (UGTs) from Arabidopsis (Arabidopsis thaliana). A ripening-related expression pattern was shown for three candidates by spatial and temporal expression analyses in five grape cultivars. Transcript accumulation correlated with the production of monoterpenyl b-D-glucosides in grape exocarp during ripening and was low in vegetative tissue. Targeted functional screening of the recombinant UGTs for their biological substrates was performed by activity-based metabolite profiling (ABMP) employing a physiologic library of aglycones built from glycosides isolated from grape. This approach led to the identification of two UDP-glucose:monoterpenol b-D-glucosyltransferases. Whereas VvGT14a glucosylated geraniol, R,S-citronellol, and nerol with similar efficiency, the three allelic forms VvGT15a, VvGT15b, and VvGT15c preferred geraniol over nerol. Kinetic resolution of R,S-citronellol and R,S-linalool was shown for VvGT15a and VvGT14a, respectively. ABMP revealed geraniol as the major biological substrate but also disclosed that these UGTs may add to the production of further glycoconjugates in planta. ABMP of aglycone libraries provides a versatile tool to uncover novel biologically relevant substrates of small-molecule glycosyltransferases that often show broad sugar acceptor promiscuity.Plant secondary metabolites are frequently decorated with Glc that is transferred from uridine diphosphoglucose by a so-called family 1 URIDINE DIPHOSPHATE GLUCOSYLTRANSFERASE (UGT; Caputi et al., 2012). Since the glucosylation process can be envisaged as a simple nucleophilic displacement reaction of Secondorder Nucleophilic Substitution type, the product is a b-glucoside. A remarkably large array of different small molecules is glucosylated in planta, including terpenoids, alkaloids, cyanohydrins, and glucosinolates, as well as flavonoids, isoflavonoids, anthocyanidins, and phenylpropanoids. In grape (Vitis vinifera) alone, more than 200 different glucosides have been identified, and there is special interest in those glucoconjugates, which can contribute to wine flavor after the hydrolytic release of volatiles during the biotechnological vinification sequence leading from grape to aged wine (Wirth et al., 2001). These flavorless glucoconjugates accumulate in grape berries during maturation and can be grouped according to their linked aglycones into monoterpenes, C13-norisoprenoids, aliphatic alcohols, and shikimatederived benzoids and phenylpropanoids (Sefton et al., 1996). It still remains an open question how this large number of structurally different flavor precursors is actually glucosylated in vivo, because the in vitro activities of single UGTs show large differences in the individual range of acceptors. Some UGTs are considered highly specific with respect to substrate ...

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The present review integrates current knowledge on mono- and sesquiterpenes in grapes with a special focus on biochemical and physiological aspects. Recent research has impressively shown the prominence of terpenoid metabolism in grapevine (Vitis sp). The 69 putatively functional mono- and sesquiterpene synthases that were identified by the analysis of the updated 12-fold sequencing and assembly of the grapevine genome deliver the scaffolds for structural diversity and display a surprising expansion of the terpene synthase (TPS) gene family in grapevine when compared to other plants like Arabidopsis thaliana (32 TPS). While monoterpenes occur as highly functionalized compounds and are stored as their corresponding glycoconjugates in berry tissues, sesquiterpenes are mainly present as unsaturated hydrocarbons and accumulate in the epicuticular wax layer of intact berries. Interestingly, both groups of terpenes appear to be involved as volatile organic compounds in plant defense and their biosynthesis is enhanced via the jasmonic acid signaling pathway. These novel aspects will help to understand how environmental cues affect the genes and enzymes of various metabolic pathways of relevant wine aroma compounds with numerous links to enology and wine flavor chemistry.