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[DNA条形码研究及其在白及属药用植物中的应用]

[DNA barcoding research and its application on medicinal plants of Bletilla H. G. Reichenbach].

作者信息

Wu Jin-Song, Zhang Yu-Si, Liu Wei, Hou Bei-Wei, Tong Wen-Jun, Zhang Li, Zhang Wei-Ming, Ding Xiao-Yu

机构信息

Nanjing Normal University, China.

出版信息

Yao Xue Xue Bao. 2014 Oct;49(10):1466-74.

Abstract

To identify adulterants from medicinal plants of Bletilla H. G. Reichenbach, the suitable candidate DNA barcoding of Bletilla was evaluated. In this study, the internal transcribed spacer (ITS) of nuclear ribosomal DNA, the LFY homologous gene intron 2 and chloroplast ycfl gene were amplified and sequenced from forty-one samples. The intra-specific and inter-specific divergences of Bletilla were calculated, and the identification efficiency was assessed using Barcoding Gap, NJ tree by K2P distance and BLAST1 method. The result showed the intra-specific divergence of nrDNA ITS and ycJfl (0.022-0.106 and 0.017-0.106) were obviously higher than the inter-specific divergence (0-0.012 and 0-0.015), and four species of Bletilla were also accurately distinguished in NJ trees. Whereas, there was no Barcoding Gap on LFY homologous gene intron 2, thus it cannot effectively identify species of Bletilla. Using NJ tree of nrDNA ITS and ycfl gene, powdery medicine and the adulterants of Bletilla were successfully unidentified. In conclusion, nrDNA ITS and ycfl can be used as a potential DNA barcoding to identify the medicinal plants in Bletilla and its adulterants. There were only three basic differences on nrDNA ITS between "Jujing baiji" and Bletilla striata of Lu'an in Anhui province, and two basic differences in ycfl. Based on morphological and molecular data, "Jujing baiji" could be recognized as the species of Bletilla striata.

摘要

为了从白及属药用植物中鉴定掺假物,对白及合适的候选DNA条形码进行了评估。本研究从41个样本中扩增并测序了核糖体DNA的内部转录间隔区(ITS)、LFY同源基因内含子2和叶绿体ycfl基因。计算了白及的种内和种间差异,并使用条形码间隙、基于K2P距离的NJ树和BLAST1方法评估了鉴定效率。结果表明,nrDNA ITS和ycJfl的种内差异(0.022 - 0.106和0.017 - 0.106)明显高于种间差异(0 - 0.012和0 - 0.015),并且在NJ树中也准确区分了4种白及。然而,LFY同源基因内含子2上没有条形码间隙,因此它不能有效地鉴定白及属植物。利用nrDNA ITS和ycfl基因的NJ树,成功鉴定了白及的粉末状药材及其掺假物。总之,nrDNA ITS和ycfl可作为潜在的DNA条形码用于鉴定白及属药用植物及其掺假物。“靖州白及”与安徽六安的白及在nrDNA ITS上只有3个碱基差异,在ycfl上有2个碱基差异。基于形态学和分子数据,“靖州白及”可被认定为白及的一个种。

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