线粒体nad1/b-c及nad5/d-e在秦艽组植物中物种鉴定意义的评价  

The identification of Sect. Cruciata(Gentiana) species using mtDNA nad1/b-c and nad5/d-e fragments

作　　者：陆佳妮[1] 赵志礼[1] 倪梁红[1] 嘎务[2] 米玛[2] LU Jia-ni;ZHAO Zhi-li;NI Liang-hong;GAAWE Dorje;MI Ma(School of Pharmacy,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Tibetan Traditional Medical College,Lhasa 850000,China)

机构地区：[1]上海中医药大学中药学院,上海201203 [2]西藏藏医学院,西藏拉萨850000

出　　处：《药学学报》2019年第1期166-172,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金青年科学基金项目(81503354);国家自然科学基金项目(81173654)

摘　　要：龙胆科龙胆属Gentiana秦艽组Sect.Cruciata多种植物作为中药秦艽及藏药解吉(■)的基原,具有重要的药用价值。在课题组前期品种整理及遗传背景分析基础上,本文分别测定秦艽组10种23个居群69份样品以及外类群椭圆叶花锚Halenia elliptica 3份样品的线粒体nad1基因第2内含子区(nad 1/b-c)以及nad5基因第4内含子区(nad5/d-e)部分序列,对其在秦艽组植物的分子系统发育分析及物种鉴定中的意义进行评价。结果显示:nad1/b-c具一粗壮秦艽G.robusta King ex Hook.f.稳定变异位点;同时,对粗茎秦艽G.crassicaulis Duthie ex Burk.及西藏秦艽G.tibetica King ex Hook.f.有一定物种分辨率;nad5/d-e序列相似度高,仅大叶秦艽G.macrophylla Pall.、黄管秦艽G.officinalis H.Smith及管花秦艽G.siphonantha Maxim.ex Kusnez.存在种内不同基因型。进而基于粗壮秦艽G.robusta King ex Hook.f.nad 1/b-c物种分子标记,设计一特异鉴定引物,建立粗壮秦艽特异性引物-PCR鉴定方法。本工作可为协同进化不完全、遗传背景复杂多样的中藏药高山基原植物DNA条形码的构建提供新思路。Gentiana section Cruciata(Gentianaceae)is a medicinally important section of herbs,including Chinese traditional medicine Gentianae Macrophyllae Radix and Tibetan herb Jieji.Here,we assess the taxonomic significance using mtDNA nad1/b-c and nad5/d-e sequence data.A total of 144 nad1/b-c and nad5/d-e sequences from11 species within Gentianaceae were obtained,including 138 sequences from 10 species within Gentiana section Cruciata and 6 sequences from Halenia elliptica(outgroup).The results showed that mtDNA nad1/b-c has specieslevel resolution within the section of Cruciata,i.e.the variable in the position 45 "C"could be used as a stable marker locus to distinguish G.robusta from other taxa;the variable in the position 352 and 353 "GA"could distinguish G.crassicaulis and G.tibetica from other taxa within the section.Intraspecies genotype variability was detected in nad1/b-c sequences of G.officinalis and G.siphonantha,respectively.These genotypes could be used as potential DNA barcode.In addition,intraspecies genotype variability was detected in nad5/d-e sequences of G.macrophylla G.officinalis and G.siphonantha,respectively.Based on the stable marker locus,a species-specific PCR protocol was developed using the primer PF to identifying G.robusta in the section.This study could expand the understanding of the diversity of mtDNA nad1/b-c and nad5/d-e in the genus Gentiana,and provide the essence for the species identification within Gentiana section Cruciata

关 键 词：线粒体DNA nad1基因 b-c内含子 龙胆属 秦艽组 DNA条形码 

分 类 号：R931[医药卫生—生药学]