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[目的]分析秦艽基原植物间不同DNA序列的差异,为秦艽药材DNA条形码的筛选和基原鉴定提供分子证据。[方法]采用PCR扩增纯化后直接测序的方法,测定大叶秦艽G.macrophylla pall.、麻花秦艽G.straminea Maxim.、粗茎秦艽G.crassicaulis Duth-ieex Burk.、小秦艽G.dahurica Fisch、黄管秦艽G.officinalis H.Smith5种植物的核糖体DNAITS、叶绿体DNA psbA-trnH核苷酸序列,并作序列同源性分析。[结果]cpDNA psbA-trnH序列长度变异范围为316-318bp,有7种不同的单倍型,单倍型间有7个变异位点,序列的GC含量为21.2%。最大简约树的聚类结果与单倍型反映的结果一致。nrDNA ITS序列长度变异范围为624～625bp。有5种不同的单倍型、单倍型间有12个变异位点,序列的GC含量为59.3%。最大简约树的聚类结果表明,小秦艽与麻花艽聚为一支,大叶秦艽与黄管秦艽聚为一支,粗茎秦艽位于聚类图的最基部。[结论]nrDNAITS序列较适合作秦艽基原植物的DNA分子鉴定。
[Objective] The aim of this study was to analyze the differences of DNA sequences among the original plants of Gentiana macrophylla and to provide molecular evidence for DNA barcoding screening and identification of Gentiana macrophylla. [Method] The PCR-amplified direct sequencing method was used to determine G. macrophylla pall., G. straminea Maxim., G. crassicaulis Duth-ieex Burk., G. dahurica Fisch , Rhizopus chinensis G.officinalis H. Smith 5 ribosomal DNAITS, chloroplast DNA psbA-trnH nucleotide sequence, and for sequence homology analysis. [Result] The length of cpDNA psbA-trnH varied from 316-318 bp in length, with 7 different haplotypes and 7 haplotypes. The GC content of the sequence was 21.2%. The clustering results of the largest parsimony tree are consistent with the results of haplotypes. The length of nrDNA ITS sequence ranged from 624 bp to 625 bp. There are five different haplotypes, 12 haplotypes and the GC content of the sequence is 59.3%. The clustering results of the largest parsimony tree showed that the genus Pinctada and Gentiana macrophylla converged into one, the genus Pinctada and Gentiana macrophylla clustered together, and the genus Gentiana was located in the most basal part of the cluster graph. [Conclusion] The nrDNA ITS sequence is more suitable for DNA molecular identification of Gentiana macrophylla plant.