应用DNA条形码技术对市场苦木药材的检测研究

DNA Barcoding for Identification of Picrasma quassioides and Its Adulterants on the Market

目的:利用DNA条形码技术对市场上中药材苦木进行检测,为其市场监管及用药安全提供保障。方法:对15份苦木药材(每份取样2个)共30个样品进行DNA提取,PCR扩增其ITS2序列并双向测序,应用CodonCode Aligner version 6.0.2软件对测序峰图进行校对拼接,将得到的所有序列与GenBank以及中药材DNA条形码鉴定系统分别进行比对,选择相似度最相近的物种在中国植物志网站上查询。在此基础上扩增其psbA-trnH序列以验证结果的可靠性。结果:市场购买的苦木样品中正品苦木Picrasma quassioides占70%、臭椿Ailanthus altissima占6.67%、板蓝Baphicacanthus cusia占13.33%、楝叶吴茱萸Tetradium glabrifolium占3.33%、柘Maclura tricuspidata占6.67%。结论:市场上苦木药材存在混伪品,DNA条形码技术能有效鉴别药材真伪。

DNA barcoding technology was employed for identifying commercial products called Picrasma quassioides on the market.This study provided a guarantee for the market supervision and drug safety.Genomic DNA（2 repeats for each sample） from 15 P.quassioides samples was extracted.The ITS2 sequences were amplified and bidirectionally sequenced.All sequences were assembled and obtained using the CodonCode Aligner version 6.0.2.All sequences were compared with sequences available in the GenBank and TCMBARCODE database.The highest similarity species were selected and searched on the website of Flora Reipublicae Popularis Sinicae.In addition,the psbA-trnH sequences were amplified to verify the reliability of the results.The results showed that P.quassioides（70%）,Ailanthus altissima（6.67%）,Baphicacanthus cusia（13.33%）,Tetradium glabrifolium（3.33%） and Madura tricuspidata（6.67%） were found in the commercial samples.It was concluded that there were some adulterants of P.quassioides on the market.DNA barcoding technology can effectively monitor the medicines from its adulterants.