基于DNA测序技术的市售地龙类药材基原调查与考证研究

DNA Sequencing to Identify Zoological Origin of Commercial Pheretima from Chinese Herbal Markets and Discussion on Its Herbal Textual Research

目的:进行地龙商品基原调查和本草考证,澄清商品地龙品种情况,提高中药质量。方法:通过随机采集安国、亳州等7大主流药材市场的地龙商品药材,进行DNA提取和COI片段PCR扩增与核酸测序,与建立的地龙参考核酸数据库进行比对;对主流本草进行调查和梳理。结果:我国商品地龙主要来源于34个物种,22%的基原物种为参环毛蚓(参状远盲蚓 Amynthas aspergillum ),22%为通俗环毛蚓(通俗腔蚓 Metaphire vulgaris ),稀见栉盲环毛蚓和威廉环毛蚓(<1%),55%的市售地龙均非2015年版《中华人民共和国药典》规定基原,而以保宁腔蚓 M.magna 为主;各市场主流商品地龙品种具有很大区别,具有显著的地域性特征;本草考证结果表明,历代本草中地龙基原并非一种,而是以具有“白颈”特征的环毛类蚯蚓入药,且历代本草、标准中多有变迁。结论:应建立合理、有效的地龙基原与质量控制方法。

Objective: To clarify the zoological of commercial pheretima (Dilong)in herbal markets and improve the quality of the traditional medicine,the zoological origin of commercial pheretima in Chinese herbal markets was identified and its herbal textual research was discussed. Methods: The commercial medicinal of pheretima samples was randomly collected from 7 major herbal markets,such as Anguo,Bozhou,and so on.The DNA was extracted,the COI fragment was amplified by PCR and the DNA sequencing was analyzed.The sequencing analysis results were compared with the reference database of pheretima and the major herbal was investigated and arranged. Results: The commercial pheretima origin from 34 species, zoological origin of 22% samples was Amynthas aspergillum ,22% was Metaphire vulgaris ,and less than 1% was M.guillelmi or A.pectinifera .Zoological origin of 55% specimens are not listed in Chinese pharmacopoeia ,and the main species origin was M.magna .The result of herbal textual research showed that pheretima was a multi-origin herbal in ancient herbal textual,the morphological characteristics of earthworms which with “white neck” was used in Chinese herbal medicines.The origin of pheretima changed several times in ancient herbal textual and standard documents. Conclusion: The study puts forward establish new reasonable,effective quality control methods for pheretima.