Han Jian-ping, Li Mei-ni, Luo Kun, Liu Mei-zi, Chen Xiao-chen, Chen Shi-lin
The Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education of PRC, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China.
Yao Xue Xue Bao. 2011 Nov;46(11):1408-12.
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-trnH, 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.
为通过DNA条形码技术从洋金花的掺伪品中鉴定出其原植物,对四种植物包括白曼陀罗、毛曼陀罗、曼陀罗和木本曼陀罗的ITS2、psbA-trnH、matK、rbcL序列进行了比较分析。四个区域(ITS2、psbA-trnH、matK、rbcL)的PCR和测序成功率分别为100%、90%、100%和85%。序列用CodonCode Aligner进行组装。计算K2P距离并通过MEGA 4.1构建NJ树。在ITS2序列中发现了30个单核苷酸多态性,在psbA-trnH基因间区域发现了33个插入/缺失。ITS2和psbA-trnH的种间K2P距离明显高于种内K2P距离。对于matK和rbcL,种间和种内距离之间不存在“条形码间隙”。分别构建了四个区域/组合的NJ树。木本曼陀罗的样本聚为一个分支,曼陀罗属的其他物种形成另一个分支。结果表明,ITS2或psbA-trnH均可用于从掺伪品中鉴定洋金花。