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霍山石斛转录组分析揭示了与活性成分合成相关的假定基因。

Analysis of Dendrobium huoshanense transcriptome unveils putative genes associated with active ingredients synthesis.

机构信息

Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.

Jiangsu Province Key Laboratory of Soil and Water Conservation and Ecological Restoration, Nanjing Forestry University, Nanjing, 210037, China.

出版信息

BMC Genomics. 2018 Dec 29;19(1):978. doi: 10.1186/s12864-018-5305-6.

Abstract

BACKGROUND

Dendrobium huoshanense C.Z. Tang et S.J. Cheng is a traditional Chinese herbal medicine with high medicinal value in China. Polysaccharides and alkaloids are its main active ingredients. To understand the difference of main active ingredients in different tissues, we determined the contents of polysaccharides and alkaloids in the roots, stems and leaves of D. huoshanense. In order to explore the reasons for the differences of active ingredients at the level of transcription, we selected roots, stems and leaves of D. huoshanenese for transcriptome sequencing and pathway mining.

RESULTS

The contents of polysaccharides and alkaloids of D. huoshanense were determined and it was found that there were significant differences in different tissues. A total of 716,634,006 clean reads were obtained and 478,361 unigenes were assembled by the Illumina platform sequencing. We identified 1407 carbohydrate-active related unigenes against CAZy database including 447 glycosyltransferase genes (GTs), 818 glycoside hydrolases (GHs), 60 carbohydrate esterases (CEs), 62 carbohydrate-binding modules (CBMs), and 20 polysaccharide lyases (PLs). In the glycosyltransferases (GTs) family, 315 differential expression genes (DEGs) were identified. In total, 124 and 58 DEGs were associated with the biosynthesis of alkaloids in Dh_L vs. Dh_S and Dh_R vs. Dh_L, respectively. A total of 62 DEGs associated with the terpenoid pathway were identified between Dh_R and Dh_S. Five key enzyme genes involved in the terpenoids pathway were identified, and their expression patterns in different tissues was validated using quantitative real-time PCR.

CONCLUSIONS

In summary, our study presents a transcriptome profile of D. huoshanense. These data contribute to our deeper relevant researches on active ingredients and provide useful insights into the molecular mechanisms regulating polysaccharides and alkaloids in Dendrobium.

摘要

背景

霍山石斛(Dendrobium huoshanense C.Z. Tang et S.J. Cheng)是中国传统的中药材,具有很高的药用价值。多糖和生物碱是其主要活性成分。为了了解不同组织中主要活性成分的差异,我们测定了霍山石斛根、茎和叶中多糖和生物碱的含量。为了从转录水平探讨活性成分差异的原因,我们选择了霍山石斛的根、茎和叶进行转录组测序和通路挖掘。

结果

测定了霍山石斛中多糖和生物碱的含量,发现不同组织之间存在显著差异。通过 Illumina 平台测序共获得 716,634,006 条清洁读数,组装得到 478,361 条 unigenes。我们针对 CAZy 数据库鉴定了 1407 个与碳水化合物活性相关的 unigenes,其中包括 447 个糖基转移酶基因(GTs)、818 个糖苷水解酶(GHs)、60 个糖酯酶(CEs)、62 个碳水化合物结合模块(CBMs)和 20 个多糖裂解酶(PLs)。在糖基转移酶(GTs)家族中,鉴定出 315 个差异表达基因(DEGs)。Dh_L 与 Dh_S、Dh_R 与 Dh_L 中分别有 124 个和 58 个 DEGs 与生物碱的生物合成有关。在 Dh_R 与 Dh_S 之间共鉴定出 62 个与萜类途径相关的 DEGs。鉴定出 62 个与萜类途径相关的 DEGs。鉴定出 5 个与萜类途径相关的关键酶基因,并通过定量实时 PCR 验证了它们在不同组织中的表达模式。

结论

总之,本研究提供了霍山石斛的转录组图谱。这些数据有助于我们对活性成分进行更深入的研究,并为深入了解调控霍山石斛多糖和生物碱的分子机制提供了有用的见解。

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