北京中医药大学学报2019，Vol.42Issue(1)：63-68,6.DOI:10.3969/j.issn.1006-2157.2019.01.012
基于高通量测序法及平板法研究龟甲饮片表面真菌种类及其多样性
Species and diversity of fungi on surface of tortoiseshell decocting pieces:a study based on high-throughput sequencing and plate method
摘要
Abstract
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.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Key words
tortoiseshell/fungi/high-throughput sequencing/colonial morphology/microscopic identification, DNA barcode identification分类
医药卫生引用本文复制引用
张帆,李红霞,徐清,孟杰亲,罗雪梅,刘春生,杨瑶珺..基于高通量测序法及平板法研究龟甲饮片表面真菌种类及其多样性[J].北京中医药大学学报,2019,42(1):63-68,6.基金项目
国家中医药管理局中医药行业科研专项项目(No.201507002) （No.201507002）
