Yuan Qing-Jun, Zhang Bin, Jiang Dan, Zhang Wen-Jing, Lin Tsai-Yun, Wang Nian-He, Chiou Shu-Jiau, Huang Lu-Qi
State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
Mol Ecol Resour. 2015 Mar;15(2):358-71. doi: 10.1111/1755-0998.12296. Epub 2014 Jul 10.
DNA barcodes have been increasingly used in authentication of medicinal plants, while their wide application in materia medica is limited in their accuracy due to incomplete sampling of species and absence of identification for materia medica. In this study, 95 leaf accessions of 23 species (including one variety) and materia medica of three Pharmacopoeia-recorded species of Angelica in China were collected to evaluate the effectiveness of four DNA barcodes (rbcL, matK, trnH-psbA and ITS). Our results showed that ITS provided the best discriminatory power by resolving 17 species as monophyletic lineages without shared alleles and exhibited the largest barcoding gap among the four single barcodes. The phylogenetic analysis of ITS showed that Levisticum officinale and Angelica sinensis were sister taxa, which indicates that L. officinale should be considered as a species of Angelica. The combination of ITS + rbcL + matK + trnH-psbA performed slight better discriminatory power than ITS, recovering 23 species without shared alleles and 19 species as monophyletic clades in ML tree. Authentication of materia medica using ITS revealed that the decoction pieces of A. sinensis and A. biserrata were partially adulterated with those of L. officinale, and the temperature around 80 °C processing A. dahurica decoction pieces obviously reduced the efficiency of PCR and sequencing. The examination of two cultivated varieties of A. dahurica from different localities indicated that the four DNA barcodes are inefficient for discriminating geographical authenticity of conspecific materia medica. This study provides an empirical paradigm in identification of medicinal plants and their materia medica using DNA barcodes.
DNA条形码已越来越多地用于药用植物的鉴定,然而由于物种采样不完整以及缺乏对药材的鉴定,它们在药材中的广泛应用在准确性方面受到限制。在本研究中,收集了中国药典记载的三种当归属植物的23个物种(包括1个变种)的95份叶片样本和药材,以评估四种DNA条形码(rbcL、matK、trnH-psbA和ITS)的有效性。我们的结果表明,ITS通过将17个物种解析为无共享等位基因的单系谱系,提供了最佳的鉴别能力,并且在四个单一条形码中表现出最大的条形码间隙。ITS的系统发育分析表明,欧当归和当归是姐妹类群,这表明欧当归应被视为当归属的一个物种。ITS+rbcL+matK+trnH-psbA的组合比ITS表现出略好的鉴别能力,在最大似然树中识别出23个无共享等位基因的物种和19个单系分支。使用ITS对药材进行鉴定发现,当归和独活的饮片部分掺有欧当归的饮片,80℃左右处理白芷饮片明显降低了PCR和测序效率。对来自不同产地的两个白芷栽培变种的检测表明,这四种DNA条形码在鉴别同种药材的地理真实性方面效率低下。本研究为利用DNA条形码鉴定药用植物及其药材提供了一个实证范例。