title: Molecular Cloning, Characterization, and Functional Analysis of Acetyl-CoA C-Acetyltransferase and Mevalonate Kinase Genes Involved in Terpene Trilactone Biosynthesis from Ginkgo biloba.
abstract: Ginkgolides and bilobalide, collectively termed terpene trilactones (TTLs), are terpenoids that form the main active substance ofGinkgo biloba. Terpenoids in the mevalonate (MVA) biosynthetic pathway include acetyl-CoAC-acetyltransferase (AACT) and mevalonate kinase (MVK) as core enzymes. In this study, two full-length (cDNAs) encoding AACT (GbAACT, GenBank Accession No. KX904942) and MVK (GbMVK, GenBank Accession No. KX904944) were cloned fromG. biloba. The deduced GbAACT and GbMVK proteins contain 404 and 396 amino acids with the corresponding open-reading frame (ORF) sizes of 1215 bp and 1194 bp, respectively. Tissue expression pattern analysis revealed thatGbAACTwas highly expressed in ginkgo fruits and leaves, andGbMVKwas highly expressed in leaves and roots. The functional complementation ofGbAACTin AACT-deficientSaccharomyces cerevisiaestrainΔerg10andGbMVKin MVK-deficient strainΔerg12confirmed thatGbAACTmediated the conversion of mevalonate acetyl-CoA to acetoacetyl-CoA andGbMVKmediated the conversion of mevalonate to mevalonate phosphate. This observation indicated thatGbAACTandGbMVKare functional genes in the cytosolic mevalonate (MVA) biosynthesis pathway. AfterG. bilobaseedlings were treated with methyl jasmonate and salicylic acid, the expression levels ofGbAACTandGbMVKincreased, and TTL production was enhanced. The cloning, characterization, expression and functional analysis ofGbAACTandGbMVKwill be helpful to understand more about the role of these two genes involved in TTL biosynthesis.
authors.affiliation: College of Horticulture and Gardening, Yangtze University, Jingzhou 434025, Hubei, China. chenqwx@foxmail.com.
authors.forename: Qiangwen
authors.initials: Q
authors.lastname: Chen
authors.affiliation: College of Horticulture and Gardening, Yangtze University, Jingzhou 434025, Hubei, China. xiaolinyingxue@163.com.
authors.forename: Jiaping
authors.initials: J
authors.lastname: Yan
authors.affiliation: College of Horticulture and Gardening, Yangtze University, Jingzhou 434025, Hubei, China. 15927855989@163.com.
authors.forename: Xiangxiang
authors.initials: X
authors.lastname: Meng
authors.affiliation: College of Horticulture and Gardening, Yangtze University, Jingzhou 434025, Hubei, China. xufeng198@126.com.
authors.forename: Feng
authors.initials: F
authors.lastname: Xu
authors.affiliation: College of Horticulture and Gardening, Yangtze University, Jingzhou 434025, Hubei, China. zww8312@163.com.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: College of Horticulture and Gardening, Yangtze University, Jingzhou 434025, Hubei, China. liaoyongling@yeah.net.
authors.forename: Yongling
authors.initials: Y
authors.lastname: Liao
authors.affiliation: College of Horticulture and Gardening, Yangtze University, Jingzhou 434025, Hubei, China. qujinwang@163.com.
authors.forename: Jinwang
authors.initials: J
authors.lastname: Qu
mesh_terms: D000085:Acetates / Q000494:pharmacology; D000101:Acetyl-CoA C-Acetyltransferase / Q000235:genetics* / Q000378:metabolism*; D053898:Biosynthetic Pathways; D003001:Cloning, Molecular; D003517:Cyclopentanes / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D020441:Ginkgo biloba / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D007783:Lactones / Q000378:metabolism*; D008798:Mevalonic Acid / Q000378:metabolism; D016366:Open Reading Frames; D009928:Organ Specificity; D054883:Oxylipins / Q000494:pharmacology; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics* / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D020156:Salicylic Acid / Q000494:pharmacology
chemical_list: D000085:Acetates; D003517:Cyclopentanes; D007783:Lactones; D054883:Oxylipins; D010940:Plant Proteins; C518782:trilactone; C072239:methyl jasmonate; D000101:Acetyl-CoA C-Acetyltransferase; D017853:Phosphotransferases (Alcohol Group Acceptor); C021796:mevalonate kinase; D020156:Salicylic Acid; D008798:Mevalonic Acid
keywords: GbAACT; GbMVK; Ginkgo biloba; MeJA; SA; functional complementation