目的 以线粒体COI序列为DNA条形码,建立准确鉴定海马药材正品来源的方法。方法 通过对获得的海马药材及其混伪品21个物种211条COI序列,进行种间、种内序列变异分析、DNA Barcoding Gap分析和NJ(Neighbor-Joining,邻接法)系统发育树构建和分析。结果 海马药材正品来源线纹海马、刺海马、大海马、三斑海马和小海马(海蛆)的最小种间距离均大于最大种内距离,具有明显的DNA Barcoding Gap,在NJ系统发育树上各自聚为独立的支,能与混伪品相互区分。结论 COI序列作为DNA条形码能准确鉴定海马药材及其混伪品,对保障海马临床用药安全具有重要意义。
Abstract
OBJECTIVE To establish an accurate identification method of genuine Hippocampus based on COI sequences as DNA barcodes and analyze the relationship between the genuine and adulterants of Hippocampus. METHODS Two hundred and eleven pieces of COI sequences were collected from 21 Hippocampus species in this study. After PCR amplification and sequencing of the DNAs, the DNA barcoding gap and phylogenetic cluster analysis were carried out. RESULTS The COI sequences of the five authentic species all had obvious DNA barcoding gap. For phylogenetic cluster analysis, all of the samples were monophyletic and every species could be discriminated clearly. CONCLUSION COI is an effective DNA barcode for the identification of the genuine Hippocampus and its adulterants, and have important significance for clinical medication safety.
关键词
海马 /
混伪品 /
DNA条形码 /
COI序列 /
分子鉴定
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Key words
Hippocampus /
adulterant /
DNA barcode /
COI sequence /
molecular identification
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中图分类号：
R282
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脚注
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基金
重大新药创制国家科技重大专项“中药新药安全性检测技术与标准研究(2014ZX09304307001)”
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