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中国药用柴胡的DNA条形码分析

DNA barcoding Chinese medicinal Bupleurum.

作者信息

Chao Zhi, Zeng Weiping, Liao Jing, Liu Li, Liang Zhenbiao, Li Xiaolei

机构信息

School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China.

School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China.

出版信息

Phytomedicine. 2014 Nov 15;21(13):1767-73. doi: 10.1016/j.phymed.2014.09.001. Epub 2014 Oct 15.

Abstract

We tested 4 markers, namely nuclear internal transcribed spacer 2 (ITS2), psbA-trnH, matK, and rbcL, to evaluate these candidate DNA barcodes for distinguishing Bupleuri radix (Chaihu) from its adulterants. 51 plant samples of Bupleurum representing 19 species were collected from different areas in China. Amplification and sequencing were attempted for all the 4 candidate barcode regions, whose validity was assessed in terms of the success rate of PCR amplification and sequencing, differential intra- and inter-specific divergences, DNA barcoding gap and the ability to discriminate species. The results showed that ITS2 had the best performance in identifying Bupleurum with an identification efficiency of 73.68%, which, after combining with psbA-trnH, increased to 83.33%. We further evaluated the efficiency of ITS2 for discriminating the species of Bupleurum using a large database from GenBank, which archived data of 223 samples from 74 species, and ITS2 successfully discriminated 64.13% of the samples at the species level. In conclusion, the ITS2 can serve as a potentially useful barcode for Bupleurum species, with psbA-trnH as a supplementary locus.

摘要

我们测试了4个标记,即核糖体DNA内转录间隔区2(ITS2)、psbA-trnH、matK和rbcL,以评估这些候选DNA条形码用于区分柴胡及其伪品的能力。从中国不同地区收集了代表19个物种的51份柴胡属植物样本。对所有4个候选条形码区域进行了扩增和测序,并根据PCR扩增和测序成功率、种内和种间差异、DNA条形码间隙以及区分物种的能力来评估其有效性。结果表明,ITS2在鉴定柴胡属植物方面表现最佳,鉴定效率为73.68%,与psbA-trnH联合使用后,鉴定效率提高到83.33%。我们进一步利用GenBank中的一个大型数据库评估了ITS2区分柴胡属物种的效率,该数据库存档了来自74个物种的223个样本的数据,ITS2在物种水平上成功区分了64.13%的样本。总之,ITS2可作为柴胡属物种潜在有用的条形码,psbA-trnH作为补充位点。

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