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DNA条形码鉴定洋金花及其伪品

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为建立有毒中药洋金花及其伪品DNA条形码鉴定方法,采用国际通用的条形码序列ITS2、psbA-trnH、matK和rbcL对洋金花原植物白花曼陀罗及其伪品毛曼陀罗、曼陀罗和木本曼陀罗4个种共计20份材料进行了比较研究.PCR及测序成功率分别为ITS2 (100%)、mark (100%)、psbA-trnH (90%)、rbcL (85%).采用CodonCode Aligner进行序列拼接,采用MEGA 4.1计算白花曼陀罗及其伪品的种内、种间的K2P距离,并基于K2P模型构建NJ树.结果显示ITS2序列共有30个单核苷酸多态性(SNPs)位点、psb A-trnH序列有33个单碱基的插入和缺失.ITS2和psbA-trnH序列种间遗传距离大于种内,matK和rbcL种内和种间没有明显“BarcodingGap”.4个条形码序列及其组合获得的分子系统树(ITS2、psb A-trnH、mark、rbcL、matK +rbcL)均分成了两大支,木本曼陀罗单聚为一支,该分子证据支持将Brugmansia提升为属水平.实验结果表明ITS2及psbA-trnH 序列可以作为洋金花及其伪品鉴定用的条形码序列.
Identification of Daturae Flos and its adulterants based on DNA barcoding technique
To identify the original plant of Daturae Flos from its adulterants by DNA barcoding, the sequences of ITS2, psbA-trnH, matK, rbcL of four species including Datura metel, Darura innoxia, Darura stramonium and Brugmansia arborea were compared and analyzed. The PCR and sequencing success rate of the four regions (ITS2, psbA-trnli, matK, rbcL) was 100%, 90%, 100% and 85%, respectively. Sequences were assembled with CodonCode Aligner. K2P distances were calculated and NJ tree was performed by MEGA 4.1. Thirty SNPs were found among ITS2 sequences, and 33 insert/deletes were found among psbA-trnH intergenic regions. The interspecific K2P distance of ITS2 and psbA-trnH was obviously higher than that of the intraspecific one. As to matK and rbcL, there was no "Barcoding Gap" existing between inter- and intra-specific distances. The NJ trees of the four regions/combinations were built separately. Samples of Brugmansia arborea were clustered into one clade, and the other species of Datura L. Formed another clade. The results showed that either ITS2 or psbA-trnH was useful to identify Daturae Flos from its adulterants.

Datura metelDNA barcodingmolecular identification

韩建萍、李美妮、罗焜、刘美子、陈晓辰、陈士林

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中国医学科学院、北京协和医学院药用植物研究所,中草药物质基础与资源利用教育部重点实验室,北京100193

西北农林科技大学生命科学学院,陕西杨凌712100

曼陀罗属 DNA条形码 分子鉴定

国家自然科学基金

81001608

2011

药学学报
中国药学会 中国医学科学院药物研究所

药学学报

CSTPCDCSCD北大核心
影响因子:1.274
ISSN:0513-4870
年,卷(期):2011.46(11)
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