市售动物药材僵蚕的DNA条形码鉴定研究  

Identification of Bombyx Batryticatus based on DNA barcoding technology

作　　者：贾静 石林春 姚辉 宋经元[1,2] 陈士林[1,3] 

机构地区：[1]中国医学科学院、北京协和医学院药用植物研究所,中草药物质基础与资源利用教育部重点实验室,北京100193 [2]重庆市药物种植研究所,重庆408435 [3]中国中医科学院中药研究所,北京100700

出　　处：《药学学报》2016年第11期1784-1790,共7页Acta Pharmaceutica Sinica

基　　金：公益性行业科研专项资助项目(201507002-4-1-2)

摘　　要：本研究基于DNA条形码技术对市售动物药材僵蚕进行分子鉴定研究,应用二维DNA条形码技术,建立跨平台信息交换的"互联网+"中药材物种鉴定体系。僵蚕药材同时包含动物家蚕和真菌白僵菌,采用植物基因组DNA提取试剂盒能成功扩增白僵菌ITS序列,未能扩增家蚕COI序列,而动物基因组DNA提取试剂盒提取总DNA均能成功扩增COI序列和ITS序列,并且ITS序列扩增、测序结果与植物基因组DNA提取试剂盒相一致。COI序列经鉴定为家蚕Bombyx mori Linnaeus,ITS序列经鉴定主要来源为白僵菌Beauveria bassiana(Bals.)Vuillant,少数为杂菌。基于ITS序列所构建的NJ树结果显示,白僵菌及其他致病菌株均可明显区分,且市售僵蚕药材混伪品含杂菌致病菌株。研究表明利用动物基因组DNA提取试剂盒可一次性将家蚕和白僵菌的DNA同时提取出来,有效解决同时含有动物和真菌类药材的基原鉴定。基于COI序列和ITS序列的DNA条形码分子技术能稳定、准确鉴别动物药材僵蚕,"互联网+"二维DNA条形码体系能有效监管药材流通领域的各个环节,有利于推动中药材市场的标准化和规范化。To identify the commercial medicinal materials of Bombyx Batryticatus, two-dimensional DNA barcode was used to construct the ＂Internet Plus＂ identification system for Chinese medicine, which should benefit the cross-platform communication of DNA barcode information. Bombyx Batryticatus contained Bombyx mori Linnaeus and Beauveria bassiana （Bals.） Vuillant. Both COI and ITS sequences were obtained via PCR amplification for total genomic DNA extracted from raw materials using the animal genomic DNA kit, while only ITS but no COl sequences was obtained when using the plant genomic DNA kit. The ITS sequences obtained using the animal genomic DNA kit were consistent with those using plant genomic DNA kit. The medicinal materials yielded COI sequences and identified as B. mori. According to analysis of ITS sequences, the main species of the medicinal materials were identified as B. bassiana and few were identified as other fungi. NJ trees analysis based on ITS sequences suggests that it can be easily distinguished from other fungi. Our results showed that total genomic DNA of B. mori and animal genomic DNA kit, which could effectively solve fungi mixture materials. COI and ITS regions as DNA Batryticatus. The ＂Internet Plus＂ two-dimensional DNA B. bassiana was extracted simultaneously using the the problem in species identification of animal and barcodes can stably and accurately identify Bombyx barcode system will promote the standardization and normalization of Chinese medicinal materials market.

关 键 词：僵蚕 COI序列 ITS序列 分子鉴定 

分 类 号：R931[医药卫生—生药学]