摘要
目的检测龟甲饮片表面真菌种类及多样性,为龟甲饮片用药安全提供参考依据。方法从北京市零售药店收集龟甲饮片8批,采用高通量测序法对龟甲饮片表面真菌种类及真菌多样性进行预估。同时,采用平板法培养、纯化龟甲表面真菌,获得单一菌株,依据菌落形态、显微结构、DNA条形码对真菌进行鉴定。结果高通量测序法结果显示,龟甲饮片表面真菌共计2界5门21纲56目123科173属201种,主要为曲霉属(Aspergillus)、虫草属(Cordyceps)、酵母菌属(Wickerhamamyces)、枝孢霉属(Cladosporium)、链格孢属(Alternaria)、念珠菌属(Candida)、棒杆菌属(Wallemia)、青霉属(Penicillium)。平板法检测结果显示,龟甲饮片表面真菌共计42株10种,主要为草酸青霉菌(Penicillium oxalicum)、烟曲霉(Aspergillus fumigates)、黑曲霉(Asper-gillus niger)、阿曲霉(Aspergillus amstelodami)、青霉属(Penicilliumsp.)、总状毛霉变种(Mucorracemosus f.racemosus)、白曲菌(Aspergillus candidus)、散囊菌属(Eurotiumsp.)。结论平板法可对中药饮片表面真菌进行分离纯化,高通量测序法则快速高效,综合平板法与高通量测序法可以科学、准确、全面地鉴定龟甲表面的真菌种类,为龟甲饮片的安全使用提供检测方法。
Objective To detect the species and diversity of fungi on surface of tortoiseshell decocting pieces and provide reference for medication safety of tortoiseshell decocting pieces.Methods The decocting pieces of tortoiseshell(8 batches)were collected from retail pharmacies in Beijing,and species and diversity of fungi on surface of tortoiseshell decocting pieces were estimated by using method of high-throughput sequencing(HTS).At the same time,fungi on surface of tortoiseshell decocting pieces were cultured by using plate method,and then were purified by using top purification method for obtaining a single fungus strain.The fungi on the surface of tortoiseshell decocting pieces were identified according to colony morphology,microstructure,and DNA barcodes.Results The results of HTS showed that there were 2 lines,5 doors,21 classes,56 orders,123 families,173 genera and 201 species of fungi on surface of tortoiseshell decocting pieces summed up totally,including mainly Aspergillus,Cordyceps,Wickerhamamyces,Cladosporium,Alternaria,Candida,Wallemia and Penicillium.The results of plate method showed that there were 42 strains of 10 species of fungi on surface of tortoiseshell decocting pieces summed up totally,including mainly Penicillium oxalicum,Aspergillus fumigates,Aspergillus niger,Aspergillus amstelodami,Penicillium sp.,Mucor racemosus f.racemosus,Aspergillus candidus and Eurotium sp.Conclusion The fungi on surface of tortoiseshell decocting pieces can be separated and purified by using plate method.HTS is rapid and efficient.The combination of plate method and HTS can identify scientifically,accurately and comprehensively the species of fungi on surface of tortoiseshell decocting pieces,and provide a detection method for medication safety of tortoiseshell decocting pieces.
作者
张帆
李红霞
徐清
孟杰亲
罗雪梅
刘春生
杨瑶珺
Zhang Fan;Li Hongxia;Xu Qing;Meng Jieqin;Luo Xuemei;Liu Chunsheng;Yang Yaojun(School of Traditional Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2019年第1期63-68,共6页
Journal of Beijing University of Traditional Chinese Medicine
基金
国家中医药管理局中医药行业科研专项项目(No.201507002)~~
关键词
龟甲
真菌
高通量测序
菌落形态
显微鉴定
DNA条形码鉴定
tortoiseshell
fungi
high-throughput sequencing
colonial morphology
microscopic identification,DNA barcode identification
