基于高通量测序技术对文冠木-4味汤混合粉末鉴定研究  

Identification Study of the Mixed Powder of Wenguanmu-4-Flavored Soup Based on High-Throughput Sequencing Technology

作　　者：敖敦格日乐[1,2] 乌仁珠日哈 萨其儿 马雅茹 哈布日 包桂花 拉喜那木吉拉[1,2] Aodungerile;Wurenzhuriha;Sa Qier;Ma Yaru;Ha Buri;Bao Guihua;Laxinamujila(Mongolian Medical College,Inner Mongolia Minzu University,Tongliao 028000,China;NMPA Key Laboratory for Quality Control of Traditional Chinese Medicine(Mongolian Medicine),Tongliao 028000,China)

机构地区：[1]内蒙古民族大学蒙医药学院,内蒙古通辽028000 [2]国家药品监督管理局中药(蒙药)质量控制重点实验室,内蒙古通辽028000

出　　处：《亚太传统医药》2025年第3期63-67,共5页Asia-Pacific Traditional Medicine

基　　金：国家药品监督管理局中药(蒙药)质量控制重点实验开放基金项目(MDK2023034);内蒙古自治区教育厅中(蒙)药资源开发利用集成攻关大平台项目(KYPT23004);内蒙古自治区直属高校基本科研业务费项目(GXKY22238,GXKY22138)。

摘　　要：目的:建立基于高通量测序技术对文冠木-4味汤混合样品鉴定研究的鉴定方法。方法:采用高通量测序技术提取文冠木-4味汤混合粉末样品中的总DNA,对ITS2片段进行PCR扩增,利用Illumina MiSeq平台对DNA片段进行双端测序,最后采用FLASH、QIIME、GraPhlAn及MEGA 7.0软件对序列进行整理并聚类分析,鉴定混合粉末中的物种。结果:混合样品中得到高质量的ITS2序列共331436条,最终比对到了4个物种(文冠木、诃子、秦艽和黄柏)。采用Krona软件进行群落分类学组成的交互展示秦艽序列占有效序列总量的42%、文冠木为22%,诃子为20%、黄柏为16%。结论:以ITS2作为条形码,采用高通量测序技术可以鉴定出文冠木-4味汤混合样品中的物种,也为其混合药材的鉴定提供新思路。Objective:To establish an identification method based on high-throughput sequencing technology to study the identification of the mixed samples of Wenguanmu-4-flavored soup.Methods:The total DNA in the mixed powder sample of Wenguanmu-4-flavored soup was extracted by high-throughput sequencing technology,PCR amplified the ITS2 fragment,and the DNA fragment was sequenced at both ends using the Illumina Mi Seq platform,and then finally the sequences were sorted out and clustered by using the software of FLASH,QIIME,Gra Phl An,and MEGA 7.0,so that species in the mixed powders could be identified.Species in the mixed powder.Results:A total of 331,436 high-quality ITS2 sequences were obtained from the mixed samples,and four species(Xanthoceras sorbifolia Bunge,Terminalia chebula Retz.,Gentiana macrophμlla Pall.and Cortex Phellodendri Chinensis)were finally compared.The interaction of taxonomic composition of the communities using Krona software demonstrated that Gentiana macrophμlla Pall.sequences accounted for 42% of the valid sequences,22% for Xanthoceras sorbifolia Bunge,20% for Terminalia chebula Retz,and 16% for Cortex Phellodendri Chinensis.Conclusion:ITS2 as a barcode and high-throughput sequencing technology can identify the species in the mixed sample of Wenguanmu-4-flavored soup and provide new ideas for identifying its mixed herbs.

关 键 词：文冠木-4味汤 高通量测序技术 ITS2 混合粉末 分子鉴定 

分 类 号：R284.1[医药卫生—中药学]