摘要
[目的]黄芪作为药食两用的一味中药,其主要活性成包含皂苷,黄酮,多糖等。本文主要探究黄芪的近缘物种,同时挖掘黄芪活性成分皂苷类和黄酮类合成的关键基因并探究这些关键基因在不同年限的表达变化情况。[方法]本研究通过高通量转录组学技术对黄芪进行研究。首先利用Trinity对黄芪样本的转录组数据进行拼接以及unigenes鉴定,然后与物种库进行比对并构建系统发育进化树,明确黄芪的近缘物种。通过RSEM对unigenes进行表达定量,并利用KEGG和GSEA分析对基因表达谱进行基因功能注释找到黄芪中与皂苷类和黄酮类化合物合成的关键基因,最后通过RT-qPCR对相关的关键基因进行验证,进而明确皂苷类和黄酮类化合物合成关键基因在黄芪不同生长年限的表达情况。[结果]黄芪的近缘物种是鹰嘴豆。GSEA分析得到黄芪中皂苷类合成的关键基因为Unigene0003373和Unigene0040532,黄酮类合成的关键基因为Unigene0076081和Unigene0027187,且这4个基因的表达量在四年生黄芪里显著高于1年生黄芪。[结论]Unigene0003373、Unigene0040532、Unigene0076081和Unigene0027187在黄芪皂苷类和黄酮类合成过程中起关键作用,这为明晰黄芪中皂苷和黄酮合成的关键通路,以及优化黄芪的培育方案奠定了理论基础。
[Objective]Astragalus membranaceus is a traditional Chinese medicinal and edible herb,and contains active compo-nents such as saponins,flavonoids,and polysaccharides.This study aimed to explore the related species of A.membranaceus,and discover the key genes involved in the synthesis of saponins and flavonoids,and investigate the expression changes of these key genes at different growth stages.[Methods]High-throughput transcriptomics analysis was performed on A.membrana⁃ceus.Firstly,the transcriptome data of A.membranaceus samples were assembled and identified using Trinity,and the unige-nes were aligned with the species database to construct a phylogenetic tree to determine the related species of A.membrana⁃ceus.The expression of unigenes was quantified using RSEM,and gene function annotation of the expression profiles was per-formed using KEGG and GSEA analysis to identify the key genes involved in the synthesis of saponins and flavonoids in A.membranaceus.Finally,the expression of the relevant key genes was validated using RT-qPCR to determine their expression patterns at different growth stages of A.membranaceus.[Results]We confirmed that the related species of A.membranaceus was chickpeas.GSEA analysis identified Unigene003373 and Unigene0040532 as key genes involved in saponin synthesis,and Unigene0076081 and Unigene0027187 as key genes involved in flavonoid synthesis in A.membranaceus.The expression lev-els of these four genes were significantly higher in four-year-old A.membranaceus compared to one-year-old plants.[Conclusion]During the growth of A.membranaceus,Unigene0003373,Unigene040532,Unigene0076081,and Unigene0027187 play key roles in the synthesis of saponins and flavonoids.This finding provided a theoretical foundation for elucidating the key pathways of saponin and flavonoids synthesis in A.membranaceus and optimizing cultivation strategies for A.membranaceus.
作者
段利强
程天佑
张甜甜
高山
Duan Liqiang;Cheng Tianyou;Zhang Tiantian;Gao Shan(Shanxi Academy of Advanced Research and Innovation,Taiyuan 030032,China)
出处
《山西农业大学学报（自然科学版）》
CAS
北大核心
2023年第3期23-32,共10页
Journal of Shanxi Agricultural University(Natural Science Edition)
基金
广灵县恒山黄芪种植技术推广及道地药材基地建设(2019FP-03)。
关键词
黄芪
皂苷
黄酮
转录组
基因表达
Astragalus membranaceus
Saponins
Flavone
Transcriptomics
Gene Expression
