活血丹及其同属药用植物的分子鉴定
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摘要
目的 利用DNA条形码技术对活血丹属药用植物进行分子鉴定。方法 提取植物总DNA,对活血丹植物样本的ITS2序列进行PCR扩增和测序。从GenBank中下载了活血丹及其他同属植物的ITS2序列。所有序列经Codon Code Aligner软件拼接,Seqman软件比对分析,并采用MEGA 7计算相关数据,基于K2P模型构建NJ聚类树。结果 在NJ聚类树上活血丹属4个品种分别聚为独立分支,表现出单系性。活血丹属与同科异属品种(外类群)聚为不同的支。结论 ITS2序列作为DNA条形码可以鉴定活血丹植物品种,DNA条形码技术可为中药材活血丹基源植物及其易混品种的鉴定提供新方法。
引文
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[17] Kress WJ,Wurdack KJ,Zimmer EA,et al.Use of DNA barcodes to identify flowering plants[J].Proc Natl Acad Sci USA,2005,102(23) :8369.
[2] 张前军,杨小生,朱海燕,郝小江.活血丹属植物的化学成分及药理研究进展[J].中草药,2006,37(6):950.
[3] 王川易,郭宝林,肖培根.中药分子鉴定方法评述[J].中国中药杂志,2011,36(2):237.
[4] Coleman AW.ITS2 is a double-edged tool for eukaryote evolutionary comparisons[J].Trends Genet,2003,19(7) :370.
[5] Chen SL,Yao H,Han JP,et al.Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plants pecies[J].PLoS One,2010,5(1) :e8613.
[6] 罗焜,陈士林,陈科力,等.基于芸香科的植物通用DNA条形码研究[J].中国科学 (C辑):生命科学,2010,40:342.
[7] Gao T,Yao H,Song J Y,et al.Identification of medicinal plants in the family Fabaceae using a potential DNA barcode ITS2 [J].J Ethnopharmacol,2010,130:116.
[8] 朱英杰,陈士林,姚辉,等.重楼属药用植物DNA条形码鉴定研究[J].药学学报,2010,45(3):376.
[9] Han J,Shi L C,Yao H,et al.Testing potential DNA barcoding regions in the Labiatae medicinal plants[J].Planta Med,2009,75:407.
[10] 李妮,陈士林,刘义梅,等.葫芦科植物通用DNA条形码的筛选[J].中草药,2011,42(7):1396.
[11] 刘美子,宋经元,罗焜,等.DNA条形码序列对9种蒿属药用植物的鉴定[J].中草药,2012,43(7):1393.
[12] Nilsson R H,G Bok M,R yberg E,et al.A software pipeline forprocessing and identification of fungal ITS sequences[J].Source Code Biol Med,2009,4:1.
[13] Keller A,T Schleicher J,Schultz T,et al.5.8 S-28S rRNA interaction and HMM-based ITS2 annotation[J].Gene,2009,430:50.
[14] Meyer CP,Paulay G.DNA barcoding:error rates based on comprehensive sampling[J].PLoS Biol,2005,3(12) :e422.
[15] Moritz C,Cicero C.DNA barcoding:promise and pitfalls[J].PLoS Biol,2004,2(10) :e354.
[16] Lahaye R,van der Bank M,Bogarin D,et al.DNA barcoding the floras of biodiversity hotspots[J].Proc Natl Acad Sci USA,2008,105(8) :2923.
[17] Kress WJ,Wurdack KJ,Zimmer EA,et al.Use of DNA barcodes to identify flowering plants[J].Proc Natl Acad Sci USA,2005,102(23) :8369.