Identification and characterization of a key LcTPS in the biosynthesis of volatile monoterpenes and sesquiterpenes in Litchi fruit

Litchi (Litchi chinensis Sonn.) has a desirable sweet taste and exotic aroma, making it popular in the markets. However, the biosynthesis of aroma volatiles in litchi fruit has rarely been investigated. In this study, the content and composition of volatile compounds were determined during litchi fruit ripening. In the mature green and mature red stages of litchi, 49 and 45 volatile compounds were detected, respectively. Monoterpenes were found to be the most abundant volatile compounds in mature red fruit, and their contents significantly increased compared to green fruit, mainly including citronellol, geraniol, myrcene, and D-limonene, which contributed to the aroma in litchi fruit. By comparing the expression profiles of the genes involved in the terpene synthesis pathway during fruit development, a terpene synthesis gene (LcTPS1-2) was identified and characterized as a major player in the synthesis of monoterpenes and sesquiterpenes. A subcellular localization analysis found LcTPS1-2 to be present in the plastid and cytoplasm. The recombinant LcTPS1-2 enzyme was able to catalyze the formation of three monoterpenes, myrcene, geraniol and citral, from geranyl pyrophosphate (GPP) and to convert farnesyl diphosphate (FPP) to a sesquiterpene, caryophyllene in vitro. Transgenic Arabidopsis thaliana plants overexpressing LcTPS1-2 exclusively released one monoterpene D-limonene, and three sesquiterpenes cis-thujopsene, (E)-β-famesene and trans-β-ionone. These results indicate that LcTPS1-2 plays an important role in the production of major volatile terpenes in litchi fruit and provides a basis for future investigations of terpenoid biosynthesis in litchi and other horticultural crops.