目的 利用DNA条形码技术对管花肉苁蓉、菊花、人参药材及其加工品进行鉴定,为中药以及食品的质量检测提供了新的技术手段。方法 该研究以ITS2序列和trnL(UAA)内含子P6环序列作为参考序列,对实验样品进行DNA提取,PCR扩增和双向测序,利用相似性搜索法和软件MEGA5.0对序列进行比对分析。结果 研究表明,ITS2序列能够准确鉴定管花肉苁蓉和人参药材样本,trnL(UAA)P6序列能够在高度降解的DNA样本成功扩增,管花肉苁蓉、菊花和人参加工品的trnL(UAA)P6序列与其药材完全一致,无任何变异位点,但通过相似性搜索法无法将加工品鉴定到种的水平。结论 应用ITS2序列可以准确鉴定DNA条形码对管花肉苁蓉和人参药材,trnL(UAA)P6序列在中药以及食品的质量检测中具有很好的应用前景。
Abstract
OBJECTIVE To identify the processed products and raw materials of Cistanche tubulosa, Chrysanthemum morifolium and Panax ginseng by DNA barcoding technology based on ITS2 and P6 loop of trnL intron sequences.METHODS The genomic DNA from all the samples were extracted, amplified and sequenced. The obtained sequences were aligned using MEGA5.0 and Blast method. RESULTS The ITS2 sequences could identify Cistanche tubulosa and Panax ginseng accurately, while the P6 loop of trnL intron sequences were amplified successfully in highly degraded DNA samples. The P6 loop sequences from the processed products and raw materials of Cistanche tubulosa, Chrysanthemum morifolium and Panax ginseng were identical with those of their herbs,but processed products can not be identified to species level by the similarity search method. CONCLUSION ITS2 Sequence can be used as the suitable DNA barcodes for identification of the herbs of Cistanche tubulosa, and Chrysanthemum morifolium, meanwhile, the P6 loop of trnL intron sequence holds a bright prospect in quality inspection of traditional Chinese medicines and foods.
关键词
DNA条形码 /
trnL(UAA)P6 /
ITS2 /
鉴定
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Key words
DNA barcode /
trnL(UAA)P6 /
ITS2 /
identification
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中图分类号：
R282
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脚注
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基金
中国中医科学院自主课题(Z2013363-865)
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