文献类型：期刊文献

英文题名：Functional diversity of diterpene synthases in Aconitum plants

作者：Tian, Mei[1];Jin, Baolong[1];Chen, Lingli[1,2];Ma, Rui[3];Ma, Qing[1];Li, Xiaolin[1];Chen, Tong[1];Guo, Juan[1];Ge, Hui[4];Zhao, Xin[4];Lai, Changjiangsheng[1];Tang, Jinfu[1];Cui, Guanghong[1];Huang, Luqi[1]

第一作者：Tian, Mei

通信作者：Cui, GH[1];Huang, LQ[1]

机构：[1]China Acad Chinese Med Sci, Natl Resource Ctr Chinese Mat Med, State Key Lab Qual Ensurance & Sustainable Use Dao, Beijing 100700, Peoples R China;[2]Anhui Food & Drug Inspection & Res Inst, Inst Tradit Chinese Med, Hefei 230051, Peoples R China;[3]Henan Univ Chinese Med, Sch Pharm, Zhengzhou 450046, Henan, Peoples R China;[4]Gansu Univ Tradit Chinese Med, Lanzhou 730000, Gansu, Peoples R China

第一机构：China Acad Chinese Med Sci, Natl Resource Ctr Chinese Mat Med, State Key Lab Qual Ensurance & Sustainable Use Dao, Beijing 100700, Peoples R China

通信机构：[1]corresponding author), China Acad Chinese Med Sci, Natl Resource Ctr Chinese Mat Med, State Key Lab Qual Ensurance & Sustainable Use Dao, Beijing 100700, Peoples R China.

年份：2023

卷号：202

外文期刊名：PLANT PHYSIOLOGY AND BIOCHEMISTRY

收录：;Scopus(收录号：2-s2.0-85168941330);WOS:【SCI-EXPANDED(收录号:WOS:001068015300001)】；

基金：This work was supported by the National Key R&D Program of China (2018YFA0900600, 2020YFA0908000), the CACMS Innovation Fund (CI2021A04109), the Fundamental Research Funds for the Central public welfare research institutes (ZZ14-YQ-049, ZZXT202001), Key project at central government level: The ability to establish sustainable use of valuable Chinese medicine resources (2060302). We sincerely thank Reuben J. Peters from Iowa State University for providing the modular metabolic engineering system.

语种：英文

外文关键词：Aconitum; Diterpene synthases; Diterpenoid alkaloid biosynthesis; Divergent evolution

摘要：Members of the Aconitum genus within the Ranunculaceae family are known to accumulate a broad array of medicinal and toxic diterpenoid alkaloids (DAs). Historically, ent-copalyl diphosphate (ent-CPP) was considered the sole precursor in DAs biosynthesis. However, the recent discovery of ent-8,13-CPP synthase in A. gymnandrum Maxim., which participates in ent-atiserene biosynthesis, raises the question of whether this gene is conserved throughout the Aconitum genus. In this study, RNA sequencing and PacBio Iso-sequencing were employed to identify diterpene synthases (diTPSs) in four additional Aconitum species with distinct DA compositions. In vitro and in vivo analyses functionally characterized a diverse array of 10 class II and 9 class I diTPSs. In addition to the identification of seven class II diTPSs as ent-CPP synthases, three other synthases generating ent-8,13-CPP, 8,13CPP, and 8 & alpha;-hydroxy-CPP were also discovered. Four class I kaurene synthases-like (KSLs) were observed to react with ent-CPP to yield ent-kaurene. Three KSLs not only reacted with ent-CPP but also ent-8,13-CPP to produce entatiserene. AsiKSL2-1 was found to react with 8 & alpha;-hydroxy-CPP to produce Z-abienol and AsiKSL2-2 exhibited no activity with any of the four intermediates. This research delineates the known diterpene biosynthesis pathways in six Aconitum species and explores the highly divergent diterpene synthases within the genus, which are consistent with their phylogeny and may be responsible for the differential distribution of diterpenoid alkaloids in root and aerial parts. These findings contribute valuable insights into the diversification of diterpene biosynthesis and establish a solid foundation for future investigation into DA biosynthetic pathways in Aconitum.