Two Terpene Synthases Are Involved in Multiple Sesquiterpene Biosynthesis in the Woody Vegetable, Toona sinensis

As a special woody vegetable, Chinese toon (Toona sinensis) has a unique flavor, which is mainly formed by a combination of volatile substances. The secretion and storage of volatile odorants in plants are often carried out in trichomes. Currently, studies on the formation of T. sinensis flavor in terms of biosynthetic processes and epidermal trichome morphology are scarce. Here, we conducted a detailed analysis of the morphology, structure, and distribution of trichomes on the leaves of T. sinensis. We identified three types of trichomes: non-glandular, sessile glandular, and stalked glandular. We found that the distribution of trichomes varies greatly in the natural populations of T. sinensis, and this may be closely related to the changes in volatile components. In order to clarify the relationship between secondary metabolism and trichome formation, we integrated the metabolic analysis of volatiles with transcriptome analysis and discovered two important (Terpene Synthase) TPS genes that may be directly involved in terpene synthesis. Through the heterologous expression in tobacco and the transient expression in T. sinensis, we showed that the TPS genes can participate in the synthesis of sesquiterpenes, among which TsTPS1262 can lead to the synthesis of elemene in T. sinensis. Our study provides insights into the synthesis pathways of complex volatile components in T. sinensis and also provides a basis for flavor breeding applications.