title: Functional Divergence of Diterpene Syntheses in the Medicinal Plant Salvia miltiorrhiza.
abstract: The medicinal plant Salvia miltiorrhiza produces various tanshinone diterpenoids that have pharmacological activities such as vasorelaxation against ischemia reperfusion injury and antiarrhythmic effects. Their biosynthesis is initiated from the general diterpenoid precursor (E,E,E)-geranylgeranyl diphosphate by sequential reactions catalyzed by copalyl diphosphate synthase (CPS) and kaurene synthase-like cyclases. Here, we report characterization of these enzymatic families from S. miltiorrhiza, which has led to the identification of unique pathways, including roles for separate CPSs in tanshinone production in roots versus aerial tissues (SmCPS1 and SmCPS2, respectively) as well as the unique production of ent-13-epi-manoyl oxide by SmCPS4 and S. miltiorrhiza kaurene synthase-like2 in floral sepals. The conserved SmCPS5 is involved in gibberellin plant hormone biosynthesis. Down-regulation of SmCPS1 by RNA interference resulted in substantial reduction of tanshinones, and metabolomics analysis revealed 21 potential intermediates, indicating a complex network for tanshinone metabolism defined by certain key biosynthetic steps. Notably, the correlation between conservation pattern and stereochemical product outcome of the CPSs observed here suggests a degree of correlation that, especially when combined with the identity of certain key residues, may be predictive. Accordingly, this study provides molecular insights into the evolutionary diversification of functional diterpenoids in plants.
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.).
authors.forename: Guanghong
authors.initials: G
authors.lastname: Cui
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.).
authors.forename: Lixin
authors.initials: L
authors.lastname: Duan
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.).
authors.forename: Baolong
authors.initials: B
authors.lastname: Jin
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.).
authors.forename: Jun
authors.initials: J
authors.lastname: Qian
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.).
authors.forename: Zheyong
authors.initials: Z
authors.lastname: Xue
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.).
authors.forename: Guoan
authors.initials: G
authors.lastname: Shen
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.).
authors.forename: John Hugh
authors.initials: JH
authors.lastname: Snyder
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.).
authors.forename: Jingyuan
authors.initials: J
authors.lastname: Song
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.).
authors.forename: Shilin
authors.initials: S
authors.lastname: Chen
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.).
authors.forename: Luqi
authors.initials: L
authors.lastname: Huang
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.) xqi@ibcas.ac.cn rjpeters@iastate.edu.
authors.forename: Reuben J
authors.identifier: http://orcid.org/0000-0003-4691-8477
authors.initials: RJ
authors.lastname: Peters
authors.affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (G.C., L.D., Z.X., G.S., J.H.S., X.Q.);State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China (G.C., B.J., L.H.);Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China (J.Q., J.S., S.C.); andDepartment of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011 (R.J.P.) xqi@ibcas.ac.cn rjpeters@iastate.edu.
authors.forename: Xiaoquan
authors.identifier: http://orcid.org/0000-0001-7175-115X
authors.initials: X
authors.lastname: Qi
mesh_terms: D004224:Diterpenes / Q000737:chemistry / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant / Q000502:physiology*; D015394:Molecular Structure; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D011994:Recombinant Proteins; D027561:Salvia miltiorrhiza / Q000235:genetics / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D004224:Diterpenes; D010940:Plant Proteins; D011994:Recombinant Proteins