川甘产藏药解吉嘎保主流品种的分子生药学鉴定  

Molecular pharmacognostical identification of mainstream varieties of Tibetan medicine Jieji Gabao from Sichuan and Gansu in China

作　　者：尕让甲 倪梁红[1] 赵志礼[1] 康恒 吴靳荣[1] 嘎务 ALA Kelsang Gyab;NI Lianghong;ZHAO Zhili;KANG Heng;WU Jinrong;GAAWE Dorje(School of Pharmacy,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Mentseekhang Traditional Tibetan Hospital,Lhasa 850000,Tibet,China;Tibetan Traditional Medical College,Lhasa 850000,Tibet,China)

机构地区：[1]上海中医药大学中药学院,上海201203 [2]西藏自治区藏医院,西藏成拉萨850000 [3]西藏藏医药大学,西藏拉萨850000

出　　处：《上海中医药大学学报》2022年第6期22-29,共8页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(82073959,81173654)。

摘　　要：目的:梳理川甘产藏药“解吉嘎保”(■)主流品种,并基于DNA条形码对其进行分子生药学鉴定研究。方法:在川甘产区进行民族植物学考察,共采集11个居群55份样品标本,对其进行基原、药材性状、显微鉴定及质量评价;并基于课题组前期相关工作进一步筛选DNA分子标记,测定各样品及西藏产麻花艽与秦艽组Sect.Cruciatar 9个近缘物种的叶绿体基因组psbA-trnH、rpl20-rps12及线粒体基因组nad1 b/c序列,以龙胆科花锚属椭圆叶花锚Halenia elliptica及龙胆属Gentiana高山龙胆组Sect.Frigidat直萼龙胆Gentiana erecto-sepala为外类群,邻接法(neighbor-joining)构建系统发育树,评价各片段的系统学意义,构建DNA条形码。结果:①川甘产藏药解吉嘎保的主流品种为龙胆科龙胆属秦艽组植物麻花艽Gentiana straminea。②各样品中龙胆苦苷和马钱苷酸总含量符合《中华人民共和国药典(2020年版)》的标准。③叶绿体psbA-trnH序列可将秦艽组10个物种分为两类,麻花艽与粗壮秦艽Gentiana robusta为一类,其余8个近缘物种为另一大类;线粒体nad1 b/c序列可进一步区分麻花艽与粗壮秦艽;叶绿体rpl20-rps12序列则具有麻花艽种下不同产地间分辨率。因此,构建川甘产藏药解吉嘎保主流品种麻花艽基于叶绿体psbA-trnH、线粒体nad1 b/c、叶绿体rpl20-rps12三个片段的组合型DNA条形码。结论:本研究明确了川甘产藏药解吉嘎保的主流品种来源,并构建了其DNA条形码鉴定方法,可为解吉嘎保的基原植物鉴定、道地藏药材质量标准的提升及龙胆科植物系统发育研究等提供基础资料。Objective:To investigate the mainstream varieties of Tibetan medicine“Jieji Gaobao”from Sichuan and Gansu in China,and study the method of molecular pharmacognosy identification based on DNA barcoding.Methods:An ethnobotanical survey of“Jieji Gabao”in Sichuan and Gansu was carried out and 55 samples from 11 populations were collected.The origin,medicinal material morphological and microscopic identification as well as quality evaluation of Jieji Gaobao were carried out.Based on our previous work,the DNA molecular markers were further screened.The sequences of chloroplast genome psbA-trnH,rpl20-rps12 and mitochondrial genome nad1 b/c in each sample and 9 related species of Gentiana Sect.Cruciata.The phylogenetic tree was constructed by neighbor-joining method with Halenia elliptica(Gentianaceae)and Gentiana erecto-sepala(Gentiana Sect.Frigidat)as the outgroups.Then the phylogenetic significances of each fragment were evaluated and DNA barcodes were constructed.Results:①Gentiana straminea(Gentianaceae Gentiana)from Sichuan and Gansu is the mainstream variety of Tibetan medicine Jieji Gabao.②The total content of gentiopicroside and loganic acid in each sample meets the standard of Chinese Pharmacopoeia.③The chloroplast psbA-trnH sequence could divide the 10 species into two groups,with Gentiana straminea and Gentiana robusta in one group and the other 8 related species in another group;the mitochondrial nad1 b/c sequence could further distinguish Gentiana straminea and Gentiana robusta;the chloroplast rpl20-rps12 sequence had the resolution between different origin.Therefore,the combinational DNA barcodes based on chloroplast psbA-trnH,mitochondrial nad1 b/c,and chloroplast rpl20-rps12 were constructed for Gentiana straminea,the mainstream Tibetan medicine“Jieji Gabao”from Sichuan and Gansu.Conclusion:In this study,the origin of the mainstream varieties of Tibetan medicine Jieji Gabao from Sichuan and Gansu were clarified,and its identification method based on DNA barcoding was constructed.This stu

关 键 词：解吉嘎保 藏药 麻花艽 分子生药学 DNA条形码 

分 类 号：R282.5[医药卫生—中药学]