仿野生与平栽蒙古黄芪中黄芪甲苷含量差异的分子机制研究  

Molecular mechanism underlying difference of astragalosideⅣcontent in imitating wild and cultivated Astragalus mongholicus

作　　者：雒阳阳 张璇 张福生[1] 李慧娟 张建琴[1] 杜冠华[2] 秦雪梅[1] LUO Yang-yang;ZHANG Xuan;ZHANG Fu-sheng;LI Hui-juan;ZHANG Jian-qin;DU Guan-hua;QIN Xue-mei(Modern Research Center for Traditional Chinese Medicine,Shanxi University,Taiyuan 030006,China;Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]山西大学中医药现代研究中心,山西太原030006 [2]中国医学科学院北京协和医学院药物研究所,北京100050

出　　处：《中国中药杂志》2022年第13期3463-3474,共12页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(31770362)。

摘　　要：系统比较分析不同生长年限下仿野生与平栽蒙古黄芪中黄芪甲苷的含量差异及其生物合成相关基因的表达情况,筛选出影响上述黄芪甲苷含量差异的关键酶基因。采用高效液相色谱(HPLC)进行黄芪甲苷含量测定;基于Illumina HiSeq和PacBio高通量转录组测序已构建的蒙古黄芪转录组数据库(RNA-Seq),筛选出与上述黄芪甲苷含量差异相关的酶基因,进行荧光定量PCR(RT-qPCR)验证,并对相关酶基因的RNA-Seq与RT-qPCR结果进行相关分析和趋势分析。发现一至四年生仿野生黄芪(IWA)与一至二年生平栽黄芪(CA)中黄芪甲苷含量变化趋势与RNA-Seq、RT-qPCR基因变化趋势相同,其中二年生IWA较一年生呈下降趋势,三年生IWA较二年生呈上升趋势,四年生IWA较三年生呈下降趋势,且一年生CA高于二年生。另五年生IWA的黄芪甲苷含量高于六年生,与RNA-Seq、RT-qPCR结果不同。该文初步阐明一至四年生IWA与一至二年生CA的黄芪甲苷含量差异与其合成途径中的上中游基因(MVK、CMK、PMK、MVD、SS)紧密相关,该结果将对黄芪药材生产种植实践有积极的指导意义。The difference of astragalosideⅣcontent and the expression of its biosynthesis related genes in imitating wild Astragalus mongolicus(IWA)and cultivated A.mongolicus(CA)under different growth years were systematically compared and analyzed.Then the key enzyme genes affected the difference of astragalosideⅣcontent in the above two A.mongolicus were screened.High-perfo-rmance liquid chromatography(HPLC)was used to determine the content of astragalosideⅣin A.mongolicusunderthe above two diffe-rent growth patterns.Based on the Illumina HiSeq and PacBio high-throughput sequencing platforms,thesecond-and third-generation transcriptome sequencing(RNA-Seq)databaseof the two A.mongolicuswas constructed.The related enzyme genes in the biosynthetic pathway of astragalosideⅣwere screened and verified byquantitative reverse transcriptase polymerase chain reaction(RT-qPCR).The RNA-sequencing(RNA-Seq)and RT-qPCR data of each gene were subjected to correlation analysis and trend analysis.The results showed that the variation trend of astragalosideⅣcontentby HPLC wasthe same as that of genes by RNA-Seq and RT-qPCR in 1-4 year IWA and 1-2 year CA.The trend level of astragalosideⅣcontentwas lower in 2-year IWA than 1-year IWA.Compared with 2-year IWA,3-year IWA had an upward trend,while 4-year IWA hada downward trend versus 3-year IWA.Additionally,1-year CA had increased trendthan 2-year CA.However,the content of astragalosideⅣin 5-year IWA was higher than that of 6-year IWA,which wasinconsistent with the findings of RNA-Seq and RT-qPCR.This study preliminarily clarifiedthat the difference of astragalosideⅣcontentin 1-4 year IWA and 1-2 year CA wasclosely related to the expression of the upstream and midstream genes(MVK,CMK,PMK,MVD,SS)in the biosynthetic pathway.The results facilitate the production and planting of Radix Astragali seu Hedysari.

关 键 词：蒙古黄芪 不同生长方式 黄芪甲苷 相关性分析 趋势分析 

分 类 号：R284.1[医药卫生—中药学]