Identification and characterization of two sesquiterpene synthase genes involved in volatile-mediated defense in tea plant (Camellia sinensis)

Terpenes and their derivatives are vital components of tea aroma. Their constitution and quantity are highly important criteria for the sensory evaluation of teas. Biologically, terpenes are involved in chemical resistance of tea plant against biotic and/or abiotic stresses. The goal of this study is to identify volatile terpenes of tea plants implicated in defense against herbivores and to identify terpene synthase (TPS) genes for their biosynthesis. Upon herbivory by tea geometrid (Ectropis obliqua Prout), tea plants were found to emit two sesquiterpenes, (E, E)-alpha-farnesene and (E)-nerolidol, which were undetectable in intact tea plants. The induced emission of (E, E)-alpha-farnesene and (E)-nerolidol suggests that they function in either direct or indirect defense of tea plants against the tea geometrid. Candidate TPS genes were identified from the transcriptomes of tea plants infested by tea geometrids. Two dedicated sesquiterpene synthases, CsAFR and CsNES2, were identified. CsAFR belongs to the TPS-b Glade and can catalyze the formation of (E, E)-alpha-farnesene from (E, E)-FPP. CsNES2 belongs to the TPS-g Glade and can synthesize (E)-nerolidol using (E, E)-FPP. The two genes were also both dramatically upregulated by herbivore damage. In summary, we showed that two novel sesquiterpene synthase genes CsAFR and CsNSE2 are inducible by herbivory and responsible for the elevated emission of herbivore-induced (E, E)-alpha-farnesene and (E)-nerolidol, which are implicated in tea plant defense against herbivores.