Integrated transcriptomic and metabolomic analyses reveal regulation of terpene biosynthesis in the stems of Sindora glabra.

Sesquiterpenes are important defensive secondary metabolites that are synthesized in various plant organs. Methyl jasmonate (MeJA) plays a key role in plant defense responses and secondary metabolism. Sindora glabra Merr. ex de Wit produces abundant sesquiterpenes in its trunks, and was subjected to investigation after MeJA treatment in order to characterize the molecular mechanisms underlying the regulation of sesquiterpene biosynthesis in plant stems and further our understanding of oleoresin production in trees. A total of 14 types of sesquiterpenes in the stems of mature S. glabra trees were identified. The levels of two sesquiterpenes, α-copaene and β-caryophyllene, significantly increased after MeJA treatment. Differentially expressed genes involved in terpenoid backbone biosynthesis were significantly enriched over time, while the expression of JAZ genes involved in the jasmonic acid signaling pathway and TGA genes involved in the salicylic acid signaling pathway was significantly enriched at later time points after treatment. Two new terpene synthase genes, SgSTPS4 and SgSTPS5, were also identified. Following MeJA treatment, the expression levels of SgSTPS1, SgSTPS2 and SgSTPS4 decreased, while SgSTPS5 expression increased. The major enzymatic products of SgSTPS4 were identified as β-elemene and cyperene, while SgSTPS5 was identified as a bifunctional mono/sesquiterpene synthase that could catalyze farnesyl pyrophosphate to produce nine types of sesquiterpenes, including α-copaene and β-caryophyllene, while SgSTPS5 could also use geranyl pyrophosphate to produce geraniol. Dramatic changes in the amounts of α-copaene and β-caryophyllene in response to MeJA were correlated with transcriptional expression changes of SgSTPS5 in the wood tissues. In addition, the transcription factors MYB, NAC, ARF, WRKY, MYC, ERF and GRAS were co-expressed with terpene biosynthesis genes and might potentially regulate terpene biosynthesis. Metabolite changes were further investigated with UPLC-TOF/MS following MeJA treatment. These results contribute to the elucidation of the molecular mechanisms of terpene biosynthesis and regulation as well as to the identification of candidate genes involved in these processes.