title: Functional Diversification of Kaurene Synthase-Like Genes inIsodon rubescens.
abstract: Ent-kaurene diterpenoids are the largest group of knownIsodonditerpenoids. Among them, oridonin is accumulated in the leaves, and is the most frequently studied compound because of its antitumor and antibacterial activities. We have identified five copalyl diphosphate synthase (CPS) and six kaurene synthase-like (KSL) genes by transcriptome profiling ofIsodon rubescensleaves. An in vitro assay assigns ten of them to five different diterpene biosynthesis pathways, except IrCPS3 that has a mutation in the catalytic motif. The Lamiaceae-specific clade genes (IrCPS1 and IrCPS2) synthesize the intermediate copalyl diphosphate (normal-CPP), while IrCPS4 and IrCPS5 synthesize the intermediateent-copalyl diphosphate (ent-CPP). IrKSL2, IrKSL4, and IrKSL5 react withent-CPP to produce anent-isopimaradiene-like compound,ent-atiserene andent-kaurene, respectively. Correspondingly, the Lamiaceae-specific clade genes IrKSL1 or IrKSL3 combined with normal-CPP led to the formation of miltiradiene. The compound then underwent aromatization and oxidization with a cytochrome P450 forming two related compounds, abietatriene and ferruginol, which were detected in the root bark. IrKSL6 reacts with normal-CPP to produce isopimaradiene. IrKSL3 and IrKSL6 have the γβα tridomain structure, as these proteins tend to possess the bidomain structure of IrKSL1, highlighting the evolutionary history of KSL gene domain loss and further elucidating chemical diversity evolution from a macroevolutionary stance in Lamiaceae.
authors.affiliation: 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 (B.J., G.C., J.G., J.T., H.L., Y.S., T.C., H.Z., L.H.); and.|Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (L.D.).
authors.forename: Baolong
authors.identifier: http://orcid.org/0000-0002-3898-2228
authors.initials: B
authors.lastname: Jin
authors.affiliation: 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 (B.J., G.C., J.G., J.T., H.L., Y.S., T.C., H.Z., L.H.); and.|Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (L.D.).
authors.forename: Guanghong
authors.identifier: http://orcid.org/0000-0001-6072-7867
authors.initials: G
authors.lastname: Cui
authors.affiliation: 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 (B.J., G.C., J.G., J.T., H.L., Y.S., T.C., H.Z., L.H.); and.|Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (L.D.).
authors.forename: Juan
authors.initials: J
authors.lastname: Guo
authors.affiliation: 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 (B.J., G.C., J.G., J.T., H.L., Y.S., T.C., H.Z., L.H.); and.|Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (L.D.).
authors.forename: Jinfu
authors.initials: J
authors.lastname: Tang
authors.affiliation: 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 (B.J., G.C., J.G., J.T., H.L., Y.S., T.C., H.Z., L.H.); and.|Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (L.D.).
authors.forename: Lixin
authors.initials: L
authors.lastname: Duan
authors.affiliation: 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 (B.J., G.C., J.G., J.T., H.L., Y.S., T.C., H.Z., L.H.); and.|Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (L.D.).
authors.forename: Huixin
authors.initials: H
authors.lastname: Lin
authors.affiliation: 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 (B.J., G.C., J.G., J.T., H.L., Y.S., T.C., H.Z., L.H.); and.|Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (L.D.).
authors.forename: Ye
authors.initials: Y
authors.lastname: Shen
authors.affiliation: 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 (B.J., G.C., J.G., J.T., H.L., Y.S., T.C., H.Z., L.H.); and.|Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (L.D.).
authors.forename: Tong
authors.initials: T
authors.lastname: Chen
authors.affiliation: 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 (B.J., G.C., J.G., J.T., H.L., Y.S., T.C., H.Z., L.H.); and.|Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (L.D.).
authors.forename: Huabei
authors.initials: H
authors.lastname: Zhang
authors.affiliation: 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 (B.J., G.C., J.G., J.T., H.L., Y.S., T.C., H.Z., L.H.); and huangluqi01@126.com.|Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China (L.D.) huangluqi01@126.com.
authors.forename: Luqi
authors.initials: L
authors.lastname: Huang
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D053898:Biosynthetic Pathways; D045786:Diterpenes, Kaurane / Q000737:chemistry / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D037602:Isodon / Q000201:enzymology* / Q000235:genetics*; D058977:Molecular Sequence Annotation; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017423:Sequence Analysis, RNA; D059467:Transcriptome / Q000235:genetics
chemical_list: D045786:Diterpenes, Kaurane; D012333:RNA, Messenger; D019883:Alkyl and Aryl Transferases