基于DNA条形码和1H-NMR代谢组学技术的福建特色药材金线莲的基原鉴定与亲缘关系研究

Anoectochilus Herba is a kind of characteristic herbal medicine in Fujian province. The original plants of Anoectochilus Herba are complex and often confused with others, and the morphological identification of it is very difficult. The natural resource of Anoectochilus Herba can’t meet the demand of market, so it is urgent to set up an accurate and reliable method for identification of Anoectochilus Herba, simultaneous with obtaining an alternative resource. In this project, DNA barcoding technique was used to identify the original plant of Anoectochilus Herba and its adulterants, and screen out a proper DNA barcode that would be optimal for the identification of Anoectochilus Herba based on the results of the morphologic studies. Meanwhile, a 1H-NMR-based metabolomics protocol analysis was applied to establish the metabolic fingerprinting, analyse the differences of the secondary metabolites, and identify the potential metabolic markers of the original plant of Anoectochilus Herba and its adulterants. Finally, by combining the result of these two methods, a two-dimensional identification methods will be established. Then, under the guidance of medicinal plant relative theory, the relationship of the origin plants of Anoectochilus Herba and its adulterants in genetic material and chemical constituents will be investigated by the combination of plant taxonomy，molecular biology and metabolomics. This study may help to provide scientific basis for finding a substituable resource for Anoectochilus Herba.

金线莲Anoectochilus Herba是福建特色中草药，其来源混杂、鉴定困难，野生资源难以满足市场需求，因此亟需建立准确、可靠的鉴定方法，同时寻找金线莲药材的替代资源。本项目拟应用多片段DNA条形码鉴定技术，评价不同序列对金线莲基原植物及近缘种的鉴别能力，筛选出最适合的DNA条形码片段。同时，采用1H-NMR代谢组学技术，建立金线莲基原植物及近缘种的代谢物指纹图谱，分析金线莲基原植物和近缘种的次生代谢产物差异，明确特征代谢标志物，定量表征代谢标志物。最终，构建一种基于DNA条形码和1H-NMR代谢组学的金线莲基原植物及近缘种的二维鉴定方法。进一步在药用植物亲缘学理论指导下，结合植物分类学、分子生物学和代谢组学的研究结果，探讨金线莲基原植物及近缘种在遗传物质和化学成分间的相关性，以期为寻找金线莲药材的替代资源提供科学依据。

金线莲是福建特色中草药，其来源混杂、鉴定困难，野生资源难以满足市场需求，亟需建立准确、可靠的鉴定方法，同时寻找金线莲药材的替代资源。本项目在形态学分类的基础上，通过利用ITS2和psbA-trnH分子标记，可以对金线莲基原植物及其近缘种进行鉴别，发现金线兰属植物亲缘关系较近，金线兰属与斑叶兰属、血叶兰属、齿唇兰属植物之间亲缘关系较远；利用1H-NMR、GC-Orbitrap/MS、UPLC、HPLC等技术，建立金线莲基原植物及其近缘属的化学指纹图谱，同时对黄酮类、金线莲苷等主要活性成进行定量分析。通过化学成分分析方法，对金线莲基原植物及其近缘种进行预测分析，发现多数金线兰属植物化学成分相似，金线兰与斑叶兰属、血叶兰属、齿唇兰属植物的化学成分差异较大。本项目从形态学、分子和化学成分方面揭示了金线莲基原植物及其近缘种的内在亲缘关系，特别是活性成分在金线兰属植物中的分布规律，为金线莲药材的替代资源的开发和可持续利用奠定理论基础。