中药鉴定创新方法体系的建立与推广.pptxVIP

中药鉴定创新方法体系的建立与推广第1页/共34页 中药鉴定技术瓶颈 一万余种中药材物种鉴定 品种混乱传统四大鉴定方法体系的局限 路在何方？基原鉴定形态鉴定显微鉴定理化鉴定第2页/共34页 DNA条形码 中药鉴定领域的变革与突破经典中药鉴定方法难以满足现代需求鉴定方法缺失导致严重安全性事件 -- 关木通误当木通服用，导致急性肾功能衰竭（马兜铃酸肾病事件） --土三七误当三七服用，导致严重肝毒性Hoang et al. Science Translational Medicine, 2013, 5: 197ra102.Nortier J L, Martinez M C M, Schmeiser H H, et al. New England Journal of Medicine, 2000, 342(23): 1686-1692.关木通百度百科木通中药材天地网第3页/共34页 DNA barcoding can achieve a rapid and accurate identification for herbsA powerful tool for identifying medicinal plants and traditional Chinese medicinal materialsAbrus cantoniensis Abrus precatorius Medicinal plantsChinese herbal medicinesDNA barcoding technology uses a short and standardized gene region to identify speciesOne sequence One species第4页/共34页 课题组首次提出并验证ITS2条形码，引起国内外广泛关注 国际期刊PLoS ONE关注药用植物DNA条形码。该文提出ITS2序列为药用植物通用的条形码序列，为陆地植物DNA条形码序列筛选提出了新的视角，由此引起同行的关注和讨论。被德国专业网站选为近25年来在ITS2领域最受关注的16篇文章之一http://its2.bioapps.biozentrum.uni-wuerzburg.de/该文发表后3年引文288次第5页/共34页 首次验证 ITS2为药用植物鉴定通用条形码通过研究比较psbA-trnH, matK, rbcL, rpoC1, ycf5, ITS2 和 ITS等七个候选DNA条形码，发现ITS2是最佳条形码序列ITS2长度约230bp，易于扩增和测序，物种鉴定能力强，适合中药材等存在DNA降解材料的鉴定人参、西洋参、羌活、宽叶羌活DNA条形码鉴定方法正在纳入国家药典第6页/共34页 Relative distribution of inter-specific divergence between congeneric species and intra-specific variation for four loci. The two bars in each box represent inter-specific (above) and intra-specific (below) genetic distances.Identification of the family RosaceaeW+W-Relative Ranks, n, P valueResultITS2matKW+ = 703, W- = 0, n = 37, P ≤1.139 × 10-7ITS2 matKITS2rbcLW+ = 703, W- = 0, n = 37, P ≤ 1.137 × 10-7ITS2 rbcLITS2rpoC1W+ = 703, W- = 0, n = 37, P ≤ 1.133 × 10-7ITS2 rpoC1matKrbcLW+ = 620, W- = 46, n = 36, P ≤ 6.479 × 10-6matK rbcLmatKrpoC1W+ = 655, W- = 11, n = 36, P ≤ 4.174 × 10-7matK rpoC1rbcLrpoC1W+ = 359, W- = 106, n = 30, P ≤ 0.009rbcL rpoC1Wilcoxon signed-rank tests of inter-specific divergence among loci.ITS2 and rbcL, matK, rpoC1 were compared.For 54 samples from 43 species in 19 diverse genera, t