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基于转录组测序和代谢物谱分析的菘蓝活性化合物的生物合成。

Biosynthesis of the active compounds of Isatis indigotica based on transcriptome sequencing and metabolites profiling.

机构信息

Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.

出版信息

BMC Genomics. 2013 Dec 5;14:857. doi: 10.1186/1471-2164-14-857.

Abstract

BACKGROUND

Isatis indigotica is a widely used herb for the clinical treatment of colds, fever, and influenza in Traditional Chinese Medicine (TCM). Various structural classes of compounds have been identified as effective ingredients. However, little is known at genetics level about these active metabolites. In the present study, we performed de novo transcriptome sequencing for the first time to produce a comprehensive dataset of I. indigotica.

RESULTS

A database of 36,367 unigenes (average length = 1,115.67 bases) was generated by performing transcriptome sequencing. Based on the gene annotation of the transcriptome, 104 unigenes were identified covering most of the catalytic steps in the general biosynthetic pathways of indole, terpenoid, and phenylpropanoid. Subsequently, the organ-specific expression patterns of the genes involved in these pathways, and their responses to methyl jasmonate (MeJA) induction, were investigated. Metabolites profile of effective phenylpropanoid showed accumulation pattern of secondary metabolites were mostly correlated with the transcription of their biosynthetic genes. According to the analysis of UDP-dependent glycosyltransferases (UGT) family, several flavonoids were indicated to exist in I. indigotica and further identified by metabolic profile using UPLC/Q-TOF. Moreover, applying transcriptome co-expression analysis, nine new, putative UGTs were suggested as flavonol glycosyltransferases and lignan glycosyltransferases.

CONCLUSIONS

This database provides a pool of candidate genes involved in biosynthesis of effective metabolites in I. indigotica. Furthermore, the comprehensive analysis and characterization of the significant pathways are expected to give a better insight regarding the diversity of chemical composition, synthetic characteristics, and the regulatory mechanism which operate in this medical herb.

摘要

背景

菘蓝是一种在中医药中广泛用于治疗感冒、发热和流感的草药。已经鉴定出各种结构类别的化合物作为有效成分。然而,在遗传学水平上,对这些活性代谢物知之甚少。在本研究中,我们首次进行了从头转录组测序,为菘蓝生成了一个全面的数据集。

结果

通过转录组测序生成了一个包含 36367 个基因(平均长度=1115.67 个碱基)的数据库。基于转录组的基因注释,鉴定了 104 个覆盖吲哚、萜类和苯丙烷类一般生物合成途径中大多数催化步骤的基因。随后,研究了这些途径中涉及的基因的器官特异性表达模式及其对茉莉酸甲酯(MeJA)诱导的反应。有效苯丙烷类代谢物的代谢物谱表明,次生代谢物的积累模式与它们生物合成基因的转录大多相关。根据 UDP-依赖性糖基转移酶(UGT)家族的分析,表明菘蓝中存在几种类黄酮,并通过使用 UPLC/Q-TOF 的代谢谱进一步鉴定。此外,通过转录组共表达分析,建议将九个新的、假定的 UGT 作为黄酮醇糖基转移酶和木质素糖基转移酶。

结论

该数据库提供了一个涉及菘蓝中有效代谢物生物合成的候选基因库。此外,对重要途径的综合分析和表征有望更好地了解该药用植物中化学组成、合成特征和调控机制的多样性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1f75/3890716/9042d3460403/1471-2164-14-857-1.jpg

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