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利用高通量测序和从头转录组组装技术鉴定獐牙菜参与治疗代谢物生物合成的基因。

High-throughput sequencing and de novo transcriptome assembly of Swertia japonica to identify genes involved in the biosynthesis of therapeutic metabolites.

机构信息

Graduate School of Pharmaceutical Sciences, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba, 260-8675, Japan.

Medical Mycology Research Center, Chiba University, Chiba, Japan.

出版信息

Plant Cell Rep. 2016 Oct;35(10):2091-111. doi: 10.1007/s00299-016-2021-z. Epub 2016 Jul 4.

DOI:10.1007/s00299-016-2021-z
PMID:27378356
Abstract

Here, we report potential transcripts involved in the biosynthesis of therapeutic metabolites in Swertia japonica , the first report of transcriptome assembly, and characterization of the medicinal plant from Swertia genus. Swertia genus, representing over 170 plant species including herbs such as S. chirata, S. hookeri, S. longifolia, S. japonica, among others, have been used as the traditional medicine in China, India, Korea, and Japan for thousands of years. Due to the lack of genomic and transcriptomic resources, little is known about the molecular basis involved in the biosynthesis of characteristic key bioactive metabolites. Here, we performed deep-transcriptome sequencing for the aerial tissues and the roots of S. japonica, generating over 2 billion raw reads with an average length of 101 bps. Using a combined approach of three popular assemblers, de novo transcriptome assembly for S. japonica was obtained, yielding 81,729 unigenes having an average length of 884 bps and N50 value of 1452 bps, of which 46,963 unigenes were annotated based on the sequence similarity against NCBI-nr protein database. Annotation of transcriptome assembly resulted in the identification of putative genes encoding all enzymes from the key therapeutic metabolite biosynthesis pathways. Transcript abundance analysis, gene ontology enrichment analysis, and KEGG pathway enrichment analysis revealed metabolic processes being up-regulated in the aerial tissues with respect to the roots of S. japonica. We also identified 37 unigenes as potential candidates involved in the glycosylation of bioactive metabolites. Being the first report of transcriptome assembly and annotation for any of the Swertia species, this study will be a valuable resource for future investigations on the biosynthetic pathways of therapeutic metabolites and their regulations.

摘要

在这里,我们报告了与獐牙菜属植物獐牙菜生物合成治疗代谢物相关的潜在转录本,这是该属药用植物转录组组装和特征描述的首次报道。獐牙菜属植物有超过 170 个种,包括草药如獐牙菜、川西獐牙菜、椭圆叶獐牙菜、獐牙菜等,几千年来在中国、印度、韩国和日本一直被用作传统药物。由于缺乏基因组和转录组资源,对于参与特征关键生物活性代谢物生物合成的分子基础知之甚少。在这里,我们对獐牙菜的地上组织和根进行了深度转录组测序,生成了超过 20 亿条平均长度为 101bp 的原始reads。使用三种流行的组装器的组合方法,我们获得了獐牙菜的从头转录组组装,产生了 81729 个平均长度为 884bp 和 N50 值为 1452bp 的 unigenes,其中 46963 个 unigenes根据与 NCBI-nr 蛋白质数据库的序列相似性进行了注释。转录组组装的注释结果确定了编码关键治疗代谢物生物合成途径中所有酶的假定基因。转录丰度分析、基因本体富集分析和 KEGG 途径富集分析表明,与獐牙菜的根相比,地上组织的代谢过程上调。我们还鉴定了 37 个可能参与生物活性代谢物糖基化的 unigenes。作为任何獐牙菜属植物转录组组装和注释的首次报道,本研究将为未来对治疗代谢物生物合成途径及其调控的研究提供有价值的资源。

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