穿山甲属动物的DNA条形码及在穿山甲商品鉴定中的应用  

DNA Barcode of Pangolins and Its Application in Identification of Manis Squama Commodities

作　　者：李婵[1] 谢雪那 蔡炫[1] 张秋楠 田恩伟 晁志[1] LI Chan;XIE Xue-na;CAI Xuan;ZHANG Qiu-nan;TIAN En-wei;CHAO Zhi(School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学中医药学院

出　　处：《中国现代中药》2019年第9期1221-1228,共8页Modern Chinese Medicine

基　　金：公益性行业科研专项(201507002-4-2)

摘　　要：目的:完善基于COI序列的DNA条形码鉴定法,用于穿山甲属动物及其商品药材的分子鉴定。方法:从穿山甲属动物及穿山甲常见混伪品原动物的鳞甲、肌肉组织或蹄甲中提取DNA,扩增COI序列并双向测序,对序列进行K2P遗传距离分析、Barcoding gap检验、单倍型分析、系统发育树的聚类分析、条形码间隙自动检索法(ABGD)(Automatic barcode gap discovery)划分。用建立的DNA条形码方法对收集的35份穿山甲商品进行鉴定。结果: COI序列扩增成功,对齐后序列长度为600 bp。遗传距离分析显示,种间差异大于种内差异,物种间具有一定遗传间隔;系统发育树的聚类分析和ABGD划分能够有效鉴别穿山甲及其混伪品。对35份甲片商品进行了准确鉴定;发现近来出现较多的印度穿山甲甲片,对其进行性状描述。结论:基于COI序列的DNA条形码是鉴定穿山甲的有效手段。Objective: To improve the DNA barcoding method based on COI sequence for molecular identification of pangolin and commercial medicinal materials. Methods: DNA was extracted from the scales,muscle tissues or hoofs.COI sequences were amplified and bidirectional sequenced,and the sequences were analyzed by K2P genetic distance analysis,Barcoding gap test,haplotype analysis,phylogenetic tree clustering analysis,and ABGD (Automatic barcode gap discovery ).The established DNA barcoding method was used to identify 35 pangolin commodities. Results: The COI sequence was amplified successfully,and the aligned sequence length was 600 bp.Genetic distance analysis indicated that the difference between species was greater than the difference within species,and there was a certain genetic interval between species.Phylogenetic tree cluster analysis and ABGD classification can effectively identify pangolin and its adulterants.35 samples of pangolins were accurately identified.Identification results show that more Manis crassicaudata which characters were described appeared in market recently. Conclusion: DNA barcoding based on COI sequence is an effective method to identify pangolin.

关 键 词：穿山甲 DNA条形码 COI序列 商品鉴定 

分 类 号：R282.74[医药卫生—中药学] R284[医药卫生—中医学]