藏药解吉那保基原植物粗茎秦艽品种鉴定及种内遗传多样性分析  

DNA fingerprinting and genetic diversity in a threatened alpine medicinal plant, Gentiana crassicaulis(Gentianaceae) using AFLP

作　　者：宗粉粉 赵志礼[1] 倪梁红[1] 嘎务 刘铜华 ZONG Fen-fen;ZHAO Zhi-li;NI Liang-hong;GAAWE Dorje;LIU Tong-hua(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Tibetan Traditional Medical College,Lhasa 850000,China;Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]上海中医药大学,上海201203 [2]西藏藏医药大学,西藏拉萨850000 [3]北京中医药大学,北京100029

出　　处：《药学学报》2020年第8期1941-1950,共10页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金面上项目(81173654);国家自然科学基金青年科学基金项目(81503354);藏医药区域协同创新中心项目(2018XTCX005)。

摘　　要：藏药"解吉那保"(居群163株个体),结合模式标本查阅与形态学比较,将其主流品种之一鉴定为龙胆科龙胆属Gentiana粗茎秦艽Gentiana crassicaulis。粗茎秦艽,我国特有物种,仅在西藏、云南、四川、贵州、青海及甘肃等省区有分布,多见于海拔2100~4500 m的高山草甸、林下及林缘。应用AFLP技术,以龙胆科花锚属椭圆叶花锚Halenia elliptica及龙胆属近缘种麻花艽G.straminea为外类群,构建DNA指纹谱。从64对引物组合中筛选出12对,共扩增出315个条带,其中多态性条带254条,占比为80.63%。粗茎秦艽种内遗传多样性丰富,遗传变异主要存在于居群间(87%),居群内部遗传变异较小(13%)。非加权组平均法(unweighted pair-group method with arithmetic means,UPGMA)聚类树物种间拓扑结构与经典分类学观点相一致;物种内分为两大支:支I由西藏、甘肃、青海及四川各居群组成,支II由贵州及云南各居群组成。PCA分析及Mantel检验等均表明粗茎秦艽居群间遗传距离与空间地理距离具有显著的正相关关系。同时,结合SSR与SNP标记,探讨了四川康定县居群S1的分化问题。本工作可为藏药"解吉那保"种质资源保护、药材生药学鉴定及道地性探究等提供基础资料。Jieji Nabao is a common Tibetan herb.According to our ethnobotanical studies,one of its original plants is identified as Gentiana crassicaulis Duthie ex Burk.(Gentianaceae).Endemic to the Qinghai-Tibet Plateau,this medicinal alpine plant is a threatened species.In this study,163 individuals from 20 populations of G.crassicaulis were collected throughout its geographical range and amplified fragment length polymorphism(AFLP)was used to investigate genetic variation in this species.A cluster analysis was performed on the AFLP data with Halenia elliptica and Gentiana straminea as the outgroups.From 64 pairs of AFLP primer combinations,12 pairs were selected for amplification and a total of 315 bands were amplified,of which 254 bands were polymorphic,accounting for 80.63%.High genetic differentiation was detected between populations(87%),and low within populations(13%).The UPGMA(unweighted pair-group method with arithmetic means)tree was topologically consistent with the traditional taxonomic treatments at the species level,and the populations of G.crassicaulis were divided into two branches:one from Yunnan and Guizhou,the other from Tibet,Qinghai,Sichuan and Gansu.PCA analysis and the Mantel test showed that there was a positive correlation between genetic distance and geographical distance.In addition,combined with SSR and SNP markers within cpDNA,the genetic differentiation within the Sichuan population S1 was validated.

关 键 词：藏药 解吉那保 粗茎秦艽 扩增片段长度多态性 DNA指纹谱 

分 类 号：R931[医药卫生—生药学]