药学学报, 2014, 49(10): 1466-1474
引用本文:
吴劲松, 张宇思, 刘薇, 侯北伟, 童文君, 张礼, 张卫明, 丁小余. 白及属药用植物DNA条形码的确立及其应用[J]. 药学学报, 2014, 49(10): 1466-1474.
WU Jin-song, ZHANG Yu-si, LIU Wei, HOU Bei-wei, TONG Wen-jun, ZHANG Li, ZHANG Wei-ming, DING Xiao-yu. DNA barcoding research and its application on medicinal plants of Bletilla H. G. Reichenbach[J]. Acta Pharmaceutica Sinica, 2014, 49(10): 1466-1474.

白及属药用植物DNA条形码的确立及其应用
吴劲松1, 张宇思1, 刘薇1, 侯北伟1,2, 童文君1, 张礼1, 张卫明2, 丁小余1
1. 南京师范大学生命科学学院, 江苏 南京 210023;
2. 南京野生植物综合利用研究院, 江苏 南京 210042
摘要:
为建立白及属药用植物适合的DNA条形码,并以DNA条形码准确的鉴别白及属药用植物及其混伪品,本研究对41份样品基因组DNA的nrDNA ITS、LFY同源基因内含子2、叶绿体ycf1基因进行扩增、测序及序列分析;比较物种种内种间的变异,并基于K2P距离分析Barcoding Gap、构建NJ系统聚类树及BLAST1算法以评价不同序列的鉴定能力。结果显示,白及属物种nrDNA ITS和ycf1序列的种间变异(0.022~0.106和0.017~0.106)明显大于种内变异(0~0.012和0~0.015),NJ树也能准确地区分4个物种;而LFY同源基因内含子2序列未发现Barcoding Gap,不能有效的鉴别白及属物种。选用nrDNA ITS和ycf1构建NJ树成功鉴定了白及粉末状药材及其混伪品。结果表明,nrDNA ITS和ycf1序列可以作为白及属及其混伪品鉴定用的DNA条形码序列。市场上“巨茎白及”的nrDNA ITS序列和ycf1序列与安徽六安白及亲缘关系最近,NJ树均与白及聚为一支;基于形态及分子数据可初步将安徽“巨茎白及”归为白及属白及植物。
关键词:    白及属      巨茎白及      DNA条形码      nrDNA ITS      LFY同源基因内含子2      ycf1     
DNA barcoding research and its application on medicinal plants of Bletilla H. G. Reichenbach
WU Jin-song1, ZHANG Yu-si1, LIU Wei1, HOU Bei-wei1,2, TONG Wen-jun1, ZHANG Li1, ZHANG Wei-ming2, DING Xiao-yu1
1. College of Life Sciences, Nanjing Normal University, Nanjing 210023, China;
2. Nanjing Institute for Comprehensive Utilization of Wild Plants, Nanjing 210042, China
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 ycf1 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 ycf1 (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 ycf1 gene, powdery medicine and the adulterants of Bletilla were successfully unidentified. In conclusion, nrDNA ITS and ycf1 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 ycf1. Based on morphological and molecular data, "Jujing baiji" could be recognized as the species of Bletilla striata.
Key words:    Bletilla    Jujing baiji    DNA barcoding    nrDNA ITS    LFY homologous gene intron 2    ycf1   
收稿日期: 2014-04-21
基金项目: 科技部“十二五”平台建设计划(211100B512J)
通讯作者: 丁小余,Tel/Fax:86-25-85891605,E-mail:dingxynj@263.net
Email: dingxynj@263.net
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