Functional identification of the terpene synthase family involved in diterpenoid alkaloids biosynthesis in Aconitum carmichaelii
| Autor: | Mei Tian, Baolong Jin, Lingli Chen, Huang Luqi, Ma Rui, Biwei Yin, Jinfu Tang, Liuying Mao, Tong Chen, Haiyan Zhang, Guanghong Cui, Juan Guo, Chang-Jiang-Sheng Lai |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Subfamily
RM1-950 DNA sequencing Aconitum carmichaelii Transcriptome 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Biosynthesis Diterpene synthase otorhinolaryngologic diseases Diterpenoid alkaloids General Pharmacology Toxicology and Pharmaceutics Gene Aconitum 030304 developmental biology 0303 health sciences biology biology.organism_classification Biochemistry chemistry 030220 oncology & carcinogenesis Full-length transcriptome Original Article Therapeutics. Pharmacology Diterpene |
| Zdroj: | Acta Pharmaceutica Sinica. B Acta Pharmaceutica Sinica B, Vol 11, Iss 10, Pp 3310-3321 (2021) |
| ISSN: | 2211-3843 2211-3835 |
| Popis: | Aconitum carmichaelii is a high-value medicinal herb widely used across China, Japan, and other Asian countries. Aconitine-type diterpene alkaloids (DAs) are the characteristic compounds in Aconitum. Although six transcriptomes, based on short-read next generation sequencing technology, have been reported from the Aconitum species, the terpene synthase (TPS) corresponding to DAs biosynthesis remains unidentified. We apply a combination of Pacbio isoform sequencing and RNA sequencing to provide a comprehensive view of the A. carmichaelii transcriptome. Nineteen TPSs and five alternative splicing isoforms belonging to TPS-b, TPS-c, and TPS-e/f subfamilies were identified. In vitro enzyme reaction analysis functional identified two sesqui-TPSs and twelve diTPSs. Seven of the TPS-c subfamily genes reacted with GGPP to produce the intermediate ent-copalyl diphosphate. Five AcKSLs separately reacted with ent-CPP to produce ent-kaurene, ent-atiserene, and ent-13-epi-sandaracopimaradie: a new diterpene found in Aconitum. AcTPSs gene expression in conjunction DAs content analysis in different tissues validated that ent-CPP is the sole precursor to all DAs biosynthesis, with AcKSL1, AcKSL2s and AcKSL3-1 responsible for C20 atisine and napelline type DAs biosynthesis, respectively. These data clarified the molecular basis for the C20-DAs biosynthetic pathway in A. carmichaelii and pave the way for further exploration of C19-DAs biosynthesis in the Aconitum species. Graphical abstract A Pacbio full-length transcriptome of Aconitum carmichaelii were publicly available. Twelve diterpene synthases were functional identified to responsible for atisine, napelline type diterpene alkaloids (DAs) and ent-13-epi-sandaracopimaradie biosynthesis, respectively.Image 1 |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|