藏茵陈川西獐牙菜环烯醚萜类生物合成相关基因表达研究

发布时间：2018-08-22 08:53

【摘要】：藏茵陈是中华民族药中的瑰宝,也是藏药八珍之一,它是藏药中最具特色的用于治疗肝胆疾病、热症及血液病的常用药物,临床上用于治疗黄疸性肝炎、病毒性肝炎、胆囊炎等,具有重要的应用前景与开发利用价值。藏茵陈的资源植物种类特别多,可以作为藏茵陈使用的药用植物主要包括龙胆科獐牙菜属、花锚属、扁蕾属、喉毛花属、肋柱花属和虎耳草科虎耳草属的多种植物。川西獐牙菜是藏茵陈的基原植物之一,野生的该植物生于中国青藏高原海拔2000～5000米,主要分布于西藏、云南、四川西北部和青海西南部。而且藏茵陈川西獐牙菜的野生资源产量越来越少,药材市场对其需求量却越来越大。随着对野生藏茵陈川西獐牙菜需求量的增加以及在开花期大量的采收,野生资源正在枯竭。近年来,对藏茵陈川西獐牙菜药理药效及其有效化学成分的研究备受关注,然而在分子水平对该植物的研究较少。本文从表型性状、基因表达模式及有效药用成分含量三方面对藏茵陈川西獐牙菜进行了初步的研究,为将来开发利用该植物奠定基础。主要结论如下:(1)以藏茵陈基原植物之一川西獐牙菜为材料,对其株高、主茎节间长度及茎生叶长宽比共3个表型性状进行测量分析,发现株高和茎生叶长宽比在居群间存在极显著差异。(2)通过藏茵陈川西獐牙菜转录组数据的分析,鉴定出环烯醚萜类生物合成的3个代谢路径相关的39个转录本,可编码24种酶。对环烯醚萜类生物合成路径中相关基因在不同组织中的表达模式分析发现,各个基因在不同组织中的表达具有显著差异。对相关化合物在不同组织中含量的测定显示,不同组织中活性化合物含量与相关生物合成基因的表达模式并没有相关性。这也表明环烯醚萜类生物合成路径中基因的表达与代谢物的积累具有复杂性。(3)对青海省和四川省的川西獐牙菜中3个不同的有效药用成分含量进行了测定,这3种化合物为龙胆苦苷(Gentiopicrin)、獐牙菜苷(Sweroside)、獐牙菜苦苷(Swertiamarin),结果表明龙胆苦苷的含量最高。此外对同一药用成分在不同株之间的含量进行了方差分析,结果表明差异极显著。而且,不同地点间其含量也有差别。本研究通过表型性状、基因表达模式及有效药用成分含量对藏茵陈川西獐牙菜进行分析,丰富了藏茵陈川西獐牙菜的表型、有效药用成分含量及基因调控信息,为挖掘调控植株表型及有效药用成分相关酶基因及保护有限的资源打下基础,并为将来开发利用民族药奠定基础。
[Abstract]:Zang Yin Chen is a treasure of Chinese medicine and one of the eight treasures of Tibetan medicine. It is the most characteristic medicine used in the treatment of liver and bile diseases, fever and hematological diseases. It is used clinically to treat jaundice hepatitis and viral hepatitis. Cholecystitis has important application prospect and development value. There are many species of resource plants of Artemisia artemisiae. The medicinal plants that can be used as Artemisia include Swertia of Gentianaceae, Anchorage, Flower, Flower, Costellaria, Saxifraga and Saxifraga. Swertia Swertia is one of the basic plants of Artemisia sinensis. The wild plant is located at 2000 ~ 5000 m above sea level on the Qinghai-Xizang Plateau of China. It is mainly distributed in Tibet, Yunnan, northwest Sichuan and southwest Qinghai. The wild resource production of Swertia capillaris is less and less, but the demand of medicinal materials market is increasing. With the increase of the demand for wild Swertia capillaris and the harvest in blooming period, wild resources are being exhausted. In recent years, much attention has been paid to the pharmacological effects and effective chemical constituents of Swertia tinctoria, but little has been done on this plant at molecular level. In this paper, the phenotypic characters, gene expression patterns and content of effective medicinal components of Swertia capillaris were studied in order to lay a foundation for the development and utilization of this plant in the future. The main conclusions are as follows: (1) three phenotypic characters including plant height, main stem Internode length and stem leaf width ratio were measured and analyzed by using Swertia mustii, one of the original plants of Artemisia sinensis, as a material. It was found that the plant height and the ratio of stem and leaf length to width were significantly different among populations. (2) based on the analysis of transcriptional data of Swertia capillaris, 39 transcripts related to the metabolic pathways of iridoterpene biosynthesis were identified, which could encode 24 enzymes. By analyzing the expression patterns of genes related to the synthesis pathway of iridoid terpenes in different tissues, it was found that there were significant differences in the expression of each gene in different tissues. The determination of the content of related compounds in different tissues showed that there was no correlation between the content of active compounds in different tissues and the expression pattern of related biosynthesis genes. It also indicated that the gene expression and metabolites accumulation in the biosynthesis pathway of iridoid terpenes were complicated. (3) the contents of three different effective medicinal components in Swertia koraiensis were determined in Qinghai Province and Sichuan Province. The three compounds were gentiopicroside (Gentiopicrin), Swertia (Sweroside), (Sweroside), (Swertiamarin),. The results showed that the content of gentiopicrin was the highest. In addition, the variance analysis of the content of the same medicinal ingredient among different plants showed that the difference was very significant. Moreover, there are differences in their contents among different sites. In this study, phenotypic characters, gene expression patterns and effective medicinal components were analyzed to enrich the phenotypic, effective medicinal component content and gene regulation information of Swertia capillaris. It will lay a foundation for the regulation of plant phenotype and effective medicinal components related enzyme genes and the protection of limited resources, and lay a foundation for the development and utilization of national medicine in the future.
【学位授予单位】：中央民族大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S567.219;Q943.2

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