Fig. 3

Terpene synthases in R. molle genome and functional characterization of diterpene synthases. a The circos map displaying chromosomal localization of all TPSs in R. molle genome. Tandem duplication genes are presented by short red lines, and the inter-chromosomal segmental duplication gene pairs are connected by red lines. The colour lines in background indicate all syntenic blocks in the genome of R. molle. b Phylogenetic analysis of total 50 putative terpene synthases R. molle genome, PpCPS/KS (P. patens copalyl diphosphate synthase /kaurane synthase) was assigned as tree root. Bootstrap values greater than or equal to 70% are shown. c Construction of an engineered yeast strain for production of diterpene. To increase the production of GGPP, a single allele of ERG9 was knockout, tHMG1, ERG20 and BTS1 were overexpressed. d GC-MS analysis of yeast strains expressing RmTPS1, RmTPS2, respectively, two products were yielded in yeast strain harbouring RmTPS1, product 1 was identified as ent-copalol, the fragmentation pattern of product 2 matched with labd-13-en-8,15-diol in NIST database. GC-MS analysis of yeast strains combinational expressing of RmTPS1 + RmTPS5 and RmTPS2 + RmTPS5, 16α-hydroxy-ent-kaurane (product 3) was detected as a single new product of combinational expressing of RmTPS1 and RmTPS5. e The speculated rearrangement process from kaurane backbone to grayanane backbone. The process is involved a Wagner-Meerwein rearrangement, with the C5-C10 bond shifting to C5-C1 bond