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1、中国科技论文在线http://www.paper.edu.cn#藏药小檗皮的DNA条形码鉴定研究1232111**李艳，李海娇，何洋，吕秀梅，赖先荣，张艺，范刚（1.成都中医药大学民族医药学院，成都611137；52.成都中医药大学药学院，成都611137；3.成都中医药大学医学技术学院，成都611137）摘要：目的：应用ITS、ITS2和rbcL序列对藏药小檗皮进行DNA条形码鉴定。方法：以小檗皮药材的4种主流品种（刺红珠BerberisdictyophyllaFranch.、甘肃小檗Berberis
2、kansuensis10Schneid.、鲜黄小檗BerberisdiaphanaMaxim.、匙叶小檗BerberisvernaeSchneid.）为研究对象，提取药材的基因组DNA、扩增PCR和双向测序，获得ITS、ITS2和rbcL序列，通过对序列特征、变异位点、遗传距离、barcodinggap以及系统进化树的分析，筛选出鉴定小檗皮的最佳条形码序列。结果：所有样本均能提取到DNA，DNA片段多在200~750bp之间，三种序列的PCR扩增和测序效率均为100%。根据rbcL序列，四个品种能得到明
3、显的区分。15结论：基于rbcL序列的DNA条形码技术可较好的区别小檗皮药材的4种主流品种，研究结果为小檗皮药材的品种鉴定提供了参考和依据。关键词：藏药；小檗皮；DNA条形码；品种鉴定中图分类号：R28220IdentificationofBerberidisCortexUsingDNAbarcoding123211LIYan,LIHaijiao,HEYang,LVXiumei,LAIXianrong,ZHANGYi,FAN1Gang(1.CollegeofEthnicMedicine,ChengduU
4、niversityofTraditionalChineseMedicine,Chendou611137;252.Pharmacycollege,ChengduUniversityofTraditionalChineseMedicine,Chengdu611137;3.CollegeofMedicineandTechnology,ChengduUniversityofTraditionalChineseMedicine,Chengdu611137)Abstract:Objective:Tostudythe
5、DNAbarcodeidentificationofBerberidisCortexinITS,ITS2andrbcLsequences.Methods:FourmainspeciesofBerberis(BerberisdictyophyllaFranch.,Berberis30kansuensisSchneid.,BerberisdiaphanaMaxim.,BerberisvernaeSchneid.)wereselectedastheresearchobjects.ThegenomicDNAsw
6、ereextractedfromherbs，andthePCRsequenceswereamplifiedandbidirectionallysequenced.Byanalyzingthesequencecharacteristics,mutationsites,geneticdistance,barcodinggapandphylogenetictree,thebestbarcodesequencewasidentified.Results:Allthesamplescouldbeextracted
7、intoDNA,theDNAfragmentsweremostlybetween35200-750bp.ThesequencingsuccessrateofallthreesequenceswasIdentificationofBerberidisCortexUsingDNAbarcoding100%.AccordingtorbcLsequence,fourvarietiescanbeclearlydistinguished.Conclusion:DNAbarcodingbasedonrbcLseque
8、ncecandistinguishfourmainvarietiesofBerberis,andtheresultsprovidereferenceandevidencefortheidentificationofBerberidisCortex.Keywords:Tibetanmedicine;BerberidisCortex;DNAbarcoding;Speciesidentification40藏药小檗皮为典型的多基原药材，来源于小檗
