Age-dependent modulation of oleoresin production in the stem of Sindora glabra

Plants produce specialized metabolites in various organs which serve important functions in defense and development. However, the molecular regulatory mechanisms of oleoresin production in stems from broadleaved tree species are not fully understood. To determine whether endogenous developmental cues play a role in the regulation of oleoresin biosynthesis in tree stems, anatomy, multi-omics and molecular experiments were utilized to investigate the change of secretory structures, chemical profiles and gene expression in different ontogenetic stages of Sindora glabra tree, which accumulates copious amount of sesquiterpene-rich oleoresin in stems. The size of secretory canals and the concentration of five sesquiterpenes in Sindora stems exhibited obvious increase with plant age, from 0.5- to 20-year-old plants. Moreover, alpha-copaene and beta-copaene were found to be stem-specific sesquiterpenes. Metabolomic analysis revealed that salicylic acid highly accumulated in mature stems, but the content of triterpenes was greatly decreased. The expression of three repressors AUX/IAA, DELLA and JAZ involved in hormone signaling transduction pathways was significantly downregulated in stems of 10- and 20-year-old plants. Two key genes SgTPS3 and SgTPS5 were identified, whose expression was highly correlated with the accumulation patterns of specific sesquiterpenes and their enzymatic products were consistent with the chemical profiles in the stem. The promoters of three SgTPSs exhibiting high activity were isolated. Furthermore, we demonstrated that SgSPL15 directly interacts with SgTPS3 and SgTPS5 promoters and activates SgTPS5 expression but SgSPL15 inhibits SgTPS3 expression. In addition, SgSPL15 enhanced sesquiterpene levels by upregulating AtTPSs expression in Arabidopsis. These results suggested that sesquiterpene biosynthesis in S. glabra stem was dependent on the regulation of endogenous hormones as well as plant age, and SgSPL15 might act as a buffering factor to regulate sesquiterpene biosynthesis by targeting SgTPS genes.