DNA barcodes for the identification of Anoectochilus roxburghii and its adulterants
DNA barcodes for the identification of Anoectochilus roxburghii and its adulterants
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用于鉴定金线莲及其掺假品的 DNA 条形码
DOI：
10.1007/s00425-015-2353-x
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发表时间：
2015-11-01
期刊：
影响因子：
4.3
通讯作者：
Zhang, Lili
中科院分区：
文献类型：
--
作者：
Lv, Tongwei;Teng, Renda;Zhang, Lili
中草药外观相似，容易混淆，使传统方法鉴定变得复杂。本研究为金线莲及其混伪品的准确快速鉴别提供了一种基于DNA条码的科学方法。这项技术是对传统中草药鉴定方法的补充。通过对DNA条形码matK、PSBA-trnH和ITS2的比较，验证了ITS2序列是快速准确鉴定刺梨及其近缘种的有效标记。以刺梨及其混伪品的基因组DNA为模板，通过聚合酶链式反应和测序方法扩增出其ITS2序列。利用BLAST1和邻接树进行物种鉴定。根据ITS2序列将12个样品成功地划分为4个物种。刺梨的ITS2序列全长为253bp。刺梨的种内平均遗传距离为0.0021，显著低于刺梨及其混伪品的种间遗传距离(0.0380)。结果表明，ITS2序列能够准确、有效地区分刺梨及其混伪品。研究结果为其他中草药的分子鉴定提供了参考。
Chinese medicinal herbs have a similar appearance and are easily confused, complicating identification via traditional methods. This study provided a scientific approach, based on DNA barcoding, to accurately and rapidly identify Anoectochilus roxburghii and its adulterants. This technology complements traditional methods of identification of medicinal herbs. A comparison of the DNA barcodes matK, psbA-trnH and ITS2 was performed to verify that the ITS2 sequence is an effective marker for rapidly and accurately identifying A. roxburghii and its closely related species. Genomic DNA extracted from A. roxburghii and its adulterants were used as templates and the ITS2 sequence was amplified using PCR amplification and sequencing. Species identification was conducted using BLAST1 and neighbor-joining trees. The 12 samples were successfully classified into four species based on the ITS2 sequence. The ITS2 sequence length of A. roxburghii was 253 bp. The average intra-specific genetic distance of A. roxburghii was 0.0021, markedly lower than the inter-specific genetic distance between A. roxburghii and its adulterants (0.0380). Our findings illustrate that ITS2 sequence can accurately and efficiently distinguish A. roxburghii and its adulterants. In addition, the results provided reference for molecular identification of other Chinese herbal medicine.