ITS与叶绿体基因组多片段组合鉴定甘肃产秦艽基原植物的策略  

A strategy for identifying six species of Sect.Cruciata(Gentiana) in Gansu using DNA barcode sequences

作　　者：倪梁红[1] 赵志礼[1] 熊波[1] 嘎务[2] 米玛[2] 

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》2016年第5期821-827,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81173654)

摘　　要：对甘肃产龙胆属Gentiana秦艽组6种植物(粗茎秦艽G.crassicaulis、麻花秦艽G.straminea、大叶秦艽G.macrophylla、小秦艽G.dahurica、管花秦艽G.siphonantha及黄管秦艽G.officinalis)进行遗传多样性分析,并建立物种DNA条形码。测定各样品(包括外类群椭圆叶花锚Halenia elliptica)核糖体ITS区及叶绿体mat K、rbcL、rpoC1、trnL(UAA)intron、psb A-trnH、atp B-rbc L、trnS(GCU)-trnG(UCC)、rpl20-rps12、trnL(UAA)-trnF(GAA)共410条序列,对基因组内存在明显多样性的物种,梳理其基因型式样;进行基于大叶秦艽叶绿体DNA基因II型,小秦艽ITS区基因II型、III型、IV型序列鉴定相关物种的方法学研究。结果显示:7个叶绿体片段具有物种鉴定意义;大叶秦艽随机取6个样品,小秦艽或黄管秦艽每个样品随机取12个阳性克隆测序,实际获得相关基因型鉴定序列的概率大于90%。建立的甘肃产秦艽组植物DNA分子个性化鉴别方法可将6个物种有效区分。Located in the transition zone between the Qinghai-Tibet Plateau and the Loess Plateau,Gansu province is one of the distribution centers of Sect.Cruciata,Gentiana（Gentianaceae） in China.Six species in the section,G.crassicaulis,G.straminea,G.siphonantha,G.officinalis,G.dahurica and G.macrophylla,are native to Gansu.In this paper,samples of 6 species and Halenia elliptica（outgroup） were collected.Nuclear DNA ITS,chloroplast DNA mat K,rbcL,rpoC1,trnL（UAA） intron,psb A-trnH,atp B-rbc L,trnS（GCU）-trnG（UCC）,rpl20-rps12 and trnL（UAA）-trnF（GAA） were sequenced from these samples.Based on the sequence analyses,high intragenomic polymorphisms were detected in ITS regions of G.crassicaulis,G.straminea,G.siphonantha,G.officinalis and G.dahurica,and they showed incomplete concerted evolution.A methodological study to identifying such close-related species as G.macrophylla,G.officinalis and G.dahurica was carried out based on the special genotypes.The results showed that 7 cp DNA sequence fragments could be used to identify G.crassicaulis,G.straminea and G.siphonantha.With nr ITS genotype II,III and IV of G.dahurica,the species can be distinguished from the close-related G.officinalis using 12 cloned sequences in a sample（with statistical significance）.The cp DNA sequences of G.macrophylla were classified into two genotypes,and with genotype II,the species can be distinguished from the close-related G.officinalis and G.dahurica using 6 test samples each（with statistical significance）.Furthermore,DNA barcode sequences were determined for all 6 species in Gansu.Also,the studies provide some basic data for analyses of genetic diversity and identification of Gentiana species.

关 键 词：龙胆属 秦艽组 DNA条形码 基因型 遗传多样性 

分 类 号：R931[医药卫生—生药学]