Mao Liuying, Jin Baolong, Chen Lingli, Tian Mei, Ma Rui, Yin Biwei, Zhang Haiyan, Guo Juan, Tang Jinfu, Chen Tong, Lai Changjiangsheng, Cui Guanghong, Huang Luqi
College of Pharmacy, Shandong University of Chinese Medicine, Jinan 250355, China.
State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Acta Pharm Sin B. 2021 Oct;11(10):3310-3321. doi: 10.1016/j.apsb.2021.04.008. Epub 2021 Apr 18.
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 . Although six transcriptomes, based on short-read next generation sequencing technology, have been reported from the 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 transcriptome. Nineteen TPSs and five alternative splicing isoforms belonging to TPS-b, TPS-c, and TPS-e/f subfamilies were identified 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 -copalyl diphosphate. Five AcKSLs separately reacted with -CPP to produce -kaurene, -atiserene, and -13--sandaracopimaradie: a new diterpene found in . AcTPSs gene expression in conjunction DAs content analysis in different tissues validated that -CPP is the sole precursor to all DAs biosynthesis, with AcKSL1, AcKSL2s and AcKSL3-1 responsible for C atisine and napelline type DAs biosynthesis, respectively. These data clarified the molecular basis for the C-DAs biosynthetic pathway in and pave the way for further exploration of C-DAs biosynthesis in the species.
是一种在中国、日本和其他亚洲国家广泛使用的高价值药用植物。乌头碱型二萜生物碱（DAs）是该植物中的特征性化合物。尽管已经报道了基于短读长下一代测序技术的该物种的六个转录组，但与DAs生物合成相对应的萜类合酶（TPS）仍未确定。我们应用Pacbio全长转录本测序和RNA测序相结合的方法，全面了解该植物的转录组。鉴定出了19个TPS以及属于TPS-b、TPS-c和TPS-e/f亚家族的5个可变剪接异构体，通过酶反应分析功能鉴定出2个倍半萜TPS和12个二萜TPS。TPS-c亚家族的7个基因与GGPP反应生成中间体 - 柯巴基焦磷酸。5个AcKSL分别与 - CPP反应生成 - 贝壳杉烯、 - 阿替生烯和 - 13 - 异 - 山达海松酸：一种在该植物中发现的新二萜。不同组织中AcTPSs基因表达与DAs含量分析相结合，证实 - CPP是所有DAs生物合成的唯一前体，AcKSL1、AcKSL2s和AcKSL3 - 1分别负责atisine和napelline型DAs的生物合成。这些数据阐明了该植物中C型DAs生物合成途径的分子基础，并为进一步探索该物种中C型DAs的生物合成铺平了道路。
