基于分子标记和代谢组学技术的黄芪与红芪比较研究

Comparison between Astragalus membranaceus var. mongholicus and Hedysarum polybotrys based on ITS sequences and metabolomics

黄芪和红芪虽然为不同属植物,但在中医理论中具有相似的功效,而且红芪曾经作为黄芪的法定药材使用。本文以多序岩黄芪和蒙古黄芪为研究对象,通过聚合酶链式反应(polymerase chain reaction,PCR)扩增内部转录间隔区(internal transcribed spacer,ITS)序列再进行双向测序,获得二者的ITS/ITS2片段并分析其位点信息的异同。黄芪和红芪的ITS碱基序列对齐长度为616,其中508个位点保持一致,占总ITS碱基长度的82.47%;有103个位点存在差异,占总ITS碱基长度的16.72%。基因型决定表现型,采用1H NMR代谢组学技术对黄芪和红芪进行化学分析,找出二者所含化学成分的异同。根据核磁图谱共指认出代谢产物34个,黄芪与红芪共有代谢产物为27个,氨基酸、糖类等初级代谢产物较为相似,黄酮类、皂苷类物质差异较大。本研究从分子和化学角度证明黄芪和红芪既具有相似性,又具有一定差异性,为阐明其具有相似功效的物质基础和资源利用奠定了基础。

Astragalus membranaceus var. mongholicus and Hedysarum polybotrys belong to different genera, but have similar drug efficacy in traditional Chinese medicine theory, and H. polybotrys was used as the legal A. membranaceus var. mongholicus previously. In this study, similarities and differences between them were analyzed via their ITS/ITS2 fragments information. The ITS （internal transcribed spacer） regions were amplified using polymerase chain reaction and then sequenced in two-way. The alignment lengths of ITS regions were 616 bp, in which 508 loci were consistent, and 103 loci were different, accounting for 82.47% and 16.72% of the total ITS nucleotides in length, respectively. As genotype determines phenotype, ~HNMR-based metabolomic approach was further used to reveal the chemical similarities and differences between them. Thirty-four metabolites were identified in the 1HNMR spectra, and twenty-seven metabolites were the common components. Amino acids, carbohydrates and other primary metabolites were similar, while a large difference existed in the flavonoids and astragalosides. This study suggests that A. membranaceus var. mongholicus and H. polybotrys show similarities and differences from molecular and chemical perspectives, which has laid a foundation for elucidating the effective material basis of drug with similar efficacy and resources utilization.