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转录组探索以进一步了解尖叶山莨菪中托品烷生物碱的生物合成。

Transcriptome exploration for further understanding of the tropane alkaloids biosynthesis in Anisodus acutangulus.

作者信息

Cui Lijie, Huang Fenfen, Zhang Dasheng, Lin Yuping, Liao Pan, Zong Jie, Kai Guoyin

机构信息

Laboratory of Plant Biotechnology, College of Life and Environment Sciences, Shanghai Normal University, Shanghai, 200234, People's Republic of China.

出版信息

Mol Genet Genomics. 2015 Aug;290(4):1367-77. doi: 10.1007/s00438-015-1005-y. Epub 2015 Feb 10.

Abstract

Tropane alkaloids (TAs) such as anisodamine, anisodine, hyoscyamine and scopolamine are extensively used in clinical practice as anticholinergic agents. Anisodus acutangulus produces TAs in root tissue, and although several genes involved in scopolamine biosynthesis have been cloned, yet the biosynthetic pathway of TAs remains poorly understood. To further understand TAs biosynthesis mechanism, transcriptome analysis with deep RNA sequencing in A. acutangulus roots was performed in this study; 48 unigenes related to tropane, piperidine and pyridine alkaloid biosynthesis, 145 linked to the distribution of arginine to TAs biosynthesis, and 86 categorized to terpenoid backbone biosynthesis have been identified in pathway enrichment analyses with eukaryotic orthologous groups (KOG) and Kyoto encyclopedia of genes and genomes. Additionally, 82 unigenes annotated as cytochrome P450 family members seemed to be involved in secondary metabolism. Genes encoding littorine mutase/monooxygenase (CYP80F1), diamine oxidase (DAO), alcohol dehydrogenase (ADH) and aromatic amino acid aminotransferase (ArAT) may also play roles in TAs biosynthetic pathways. Furthermore, over 1,000 unigenes were identified as potential transcription factors of WRKY, AP2/ERF, MYB and bHLH families, which would be helpful to understand transcriptional regulation of secondary metabolite biosynthesis. These data enable novel insights into A. acutangulus transcriptome, updating the knowledge of TAs biosynthetic mechanism at molecular level.

摘要

托烷生物碱(TAs),如樟柳碱、山莨菪碱、莨菪碱和东莨菪碱,作为抗胆碱能药物在临床实践中被广泛使用。尖叶山莨菪在根组织中产生TAs,尽管已经克隆了几个参与东莨菪碱生物合成的基因,但TAs的生物合成途径仍知之甚少。为了进一步了解TAs的生物合成机制,本研究对尖叶山莨菪根进行了深度RNA测序的转录组分析;在真核直系同源组(KOG)和京都基因与基因组百科全书的通路富集分析中,已鉴定出48个与托烷、哌啶和吡啶生物碱生物合成相关的单基因、145个与精氨酸向TAs生物合成的分配相关的单基因,以及86个归类于萜类骨架生物合成的单基因。此外,82个被注释为细胞色素P450家族成员的单基因似乎参与了次生代谢。编码水苏碱变位酶/单加氧酶(CYP80F1)、二胺氧化酶(DAO)、乙醇脱氢酶(ADH)和芳香族氨基酸转氨酶(ArAT)的基因也可能在TAs生物合成途径中发挥作用。此外,超过1000个单基因被鉴定为WRKY、AP2/ERF、MYB和bHLH家族的潜在转录因子,这将有助于了解次生代谢物生物合成的转录调控。这些数据为尖叶山莨菪转录组提供了新的见解,更新了分子水平上TAs生物合成机制的知识。

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