粗茎秦艽转录组及其环烯醚萜类生物合成相关基因分析与验证  

Transcriptome analysis and exploration of genes involved in the biosynthesis of iridoids in Gentiana crassicaulis(Gentianaceae)

作　　者：康恒 赵志礼[1] 倪梁红[1] 李尉涛 赵淑娟[1] 刘铜华 KANG Heng;ZHAO Zhi-li;NI Liang-hong;LI Wei-tao;ZHAO Shu-juan;LIU Tong-hua(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Tibetan Traditional Medical College,Lhasa 850000,China;Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]上海中医药大学,上海201203 [2]西藏藏医药大学,西藏拉萨850000 [3]北京中医药大学,北京100029

出　　处：《药学学报》2021年第7期2005-2014,共10页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金面上项目(82073959);国家自然科学基金面上项目(81173654);藏医药区域协同创新中心项目(2018XTCX005);上海市卫生健康委员会中医药传承和科技创新项目(ZYCC2019012).

摘　　要：龙胆科龙胆属Gentiana秦艽组(Sect.Cruciata)粗茎秦艽Gentiana crassicaulis Duthie ex Burk.为中药秦艽及藏药“解吉那保”()基原植物之一,含有龙胆苦苷等环烯醚萜类活性成分,具有重要的药用价值。本文对粗茎秦艽转录组进行测序分析,分别构建其根、茎、叶、花转录组数据库,并对可能参与编码环烯醚萜苷类合成通路中关键酶的部分unigene进行qRT-PCR验证。结果如下:①共得到159534条unigene,平均长度为679。根据GO功能分类,可将unigene分为3大类67分支;基于KOG功能可将unigene分为25类;②KEGG代谢通路分析发现粗茎秦艽中215条unigene参与到20个次生代谢标准通路中,有305条unigene参与编码环烯醚萜苷类合成通路中的28个关键酶,且在不同部位中的表达存在差异;③qRT-PCR与RNA-Seq结果基本吻合,实验涉及的HMGS、DXS、MCS、GPPS、G10H、7-DLNGT及STR 7个基因皆为地上部位(茎、叶、花)的相对表达量高于地下部位(根)。环烯醚萜类为中药秦艽、龙胆、当药及青叶胆等药材的指标性成分及活性成分之一,本工作可为应用生物技术方法获取环烯醚萜类活性成分或其中间体,探讨药用植物次生产物累积规律、不同生态型品种的质量评价及药材道地性形成机制研究等提供基础科学资料。Gentiana crassicaulis Duthie ex Burk.in Gentiana(Sect.Cruciata),Gentianaceae,is one of the original plants of both Gentianae Macrophyllae Radix and Tibetan herb Jie-Ji Na-Bao,which contain such bioactive iridoids as gentiopicroside,loganic acid and others.In this study,based on previous work,the transcriptome of G.crassicaulis was sequenced and analyzed to construct transcriptome databases of roots,stems,leaves and flowers.qRT-PCR verification was conducted for parts of unigenes that may be key enzymes in the pathway of iridoid biosynthesis.The results are as follows:①a total of 159534 unigenes were obtained,with an average length of 679 bp.According to the functional classification of GO,unigenes can be divided into 3 categories with 67 branches.The unigenes were aligned in the KOG database and were classified into 25 categories according to function.②In the KEGG database,215 unigenes were implicated in 20 standard secondary metabolism pathways.The analysis shows that 305 unigenes encoded 28 key enzymes in the pathway of iridoid biosynthesis,and their expression in different organs is different;and③qRT-PCR was approximately consistent with RNA-Seq results.The 7 annotated unigenes identified in this study,HMGS,DXS,MCS,GPPS,G10H,7-DLNGT and STR,all had higher relative expression levels in the above-ground parts(stem,leaf and flower)than in the underground part(root).Iridoids are common active and index components of such traditional Chinese medicines as Qinjiao,Longdan,Dangyao,and Qingyedan,among others.Therefore,this work provides basic scientific data for further development including obtaining active components or intermediates through biotechnology,exploring the accumulation of effective components,evaluating the quality of different ecotype varieties,and identifying authentic biosynthesis pathways of medicinal materials.

关 键 词：粗茎秦艽 转录组 环烯醚萜类 生物合成 代谢通路 QRT-PCR 

分 类 号：R931[医药卫生—生药学]