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延胡索鳞茎中假定生物碱生物合成基因的鉴定及发育表达谱分析。

Identification and developmental expression profiling of putative alkaloid biosynthetic genes in Corydalis yanhusuo bulbs.

作者信息

Liao Dengqun, Wang Pengfei, Jia Chan, Sun Peng, Qi Jianjun, Zhou Lili, Li Xian'en

机构信息

Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing 100193, PR China.

China Pharmaceutical University, Nanjing 210009, China.

出版信息

Sci Rep. 2016 Jan 18;6:19460. doi: 10.1038/srep19460.

Abstract

Alkaloids in bulbs of Corydalis (C.) yanhusuo are the major pharmacologically active compounds in treatment of blood vessel diseases, tumors and various pains. However, due to the absence of gene sequences in C. yanhusuo, the genes involved in alkaloid biosynthesis and their expression during bulb development remain unknown. We therefore established the first transcriptome database of C. yanhusuo via Illumina mRNA-Sequencing of a RNA composite sample collected at Bulb initiation (Day 0), early enlargement (Day 10) and maturation (Day 30). 25,013,630 clean 90 bp paired-end reads were de novo assembled into 47,081 unigenes with an average length of 489 bp, among which 30,868 unigenes (65.56%) were annotated in four protein databases. Of 526 putative unigenes involved in biosynthesis o f various alkaloids, 187 were identified as the candidate genes involved in the biosynthesis of benzylisoquinoline alkaloids (BIAs), the only alkaloid type reported in C. yanhusuo untill now. BIAs biosynthetic genes were highly upregulated in the overall pathway during bulb development. Identification of alkaloid biosynthetic genes in C. yanhusuo provide insights on pathways and molecular regulation of alkaloid biosynthesis, to initiate metabolic engineering in order to improve the yield of interesting alkaloids and to identify potentially new alkaloids predicted from the transcriptomic information.

摘要

延胡索鳞茎中的生物碱是治疗血管疾病、肿瘤及各种疼痛的主要药理活性化合物。然而,由于延胡索缺乏基因序列,参与生物碱生物合成的基因及其在鳞茎发育过程中的表达仍不清楚。因此,我们通过对在鳞茎起始期(第0天)、早期膨大期(第10天)和成熟期(第30天)采集的RNA混合样本进行Illumina mRNA测序,建立了首个延胡索转录组数据库。25,013,630条90 bp的干净双端读段被从头组装成47,081个单基因,平均长度为489 bp,其中30,868个单基因(65.56%)在四个蛋白质数据库中得到注释。在526个参与各种生物碱生物合成的推定单基因中,187个被鉴定为参与苄基异喹啉生物碱(BIA)生物合成的候选基因,BIA是目前报道的延胡索中唯一的生物碱类型。BIA生物合成基因在鳞茎发育的整个途径中高度上调。延胡索生物碱生物合成基因的鉴定为生物碱生物合成途径和分子调控提供了见解,以启动代谢工程来提高目标生物碱的产量,并鉴定从转录组信息中预测的潜在新生物碱。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b91e/4726099/3ea64faa5696/srep19460-f1.jpg

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