目的 探究传统连栀矾溶液发酵炮制过程中真菌菌群多样性和优势菌群，为科学发酵提供理论依据。方法 采用Illumina HiSeq高通量测序技术测定不同发酵时间上下层连栀矾溶液中真菌ITS2区序列，应用QⅡME、Mothur和R等软件整理和统计样品序列数目和操作分类单元（OTU）数量，分析样品中真菌菌群的组成、丰度、分布、alpha多样性、beta多样性，阐明炮制过程中真菌菌群结构、多样性及丰富度的动态变化。结果 4个不同的发酵时期共获得用于分析的有效序列数为207 068，稀疏曲线表明测序深度充分，OTUs的数量接近于饱和，能充分展示连栀矾溶液的真菌群落结构，其主要由子囊菌门（Ascomycota，74.21%）、担子菌门（Basidiomycota，7.31%）、罗兹菌门（Rozellomycota，2.62%）、接合菌门（Zygomycota，0.66%）和球囊菌门（Glomeromycota，0.01%）组成，优势菌门为子囊菌门。通过分析OTUs数量、chao1指数，ACE指数、Shannon指数，表明连栀矾溶液真菌菌群有较高的丰富度和多样性，并且在炮制过程中上下层总丰富度和多样性随发酵时间的增加逐渐增加，其中上层样品的丰富度、多样性逐渐增加，下层样品的丰富度、多样性先降低后增加，且上层样品的丰富度、多样性明显高于下层样品。结论 以高通量测序技术分析连栀矾溶液发酵炮制过程中真菌菌群结构、多样性及丰富度的动态变化可为指导其科学发酵提供参考。

Objective To investigate the dynamic changes of fungal communitiy diversity and dominant fungal communitiy of Lianzhifan solution in the fermentation process, so as to provide a scientific basis for the scientific fermentation. Methods Different samples was furtherly measured by Illumina HiSeq sequencing technique for the sequence of ITS2 variable region of fungi from Lianzhifan solution. QⅡME, Mothur, and R software programs were employed to sort and calculate the number of sequences and operational taxonomic units (OTUs) for each sample. The composition, abundance, distribution, alpha diversity index of species, and beta diversity were analyzed to clarify the dynamic changes of fungal communities in structure, diversity, and abundance during the process of fermentation. Results The numbers of effective sequences for four different fermentation periods were 207 068; The rarefaction curves showed a sufficient sequencing depth, and the number of OTUs was close to saturation, which can fully display the fungal community structure of the Lianzhifan solution. The Fungi in Lianzhifan solution belonged to the following five phylums:Ascomycota (74.21%), Basidiomycota (7.31%), Rozellomycota (2.62%), Zygomycota (0.66%), and Glomeromycota (0.01%). The dominant specie was identified to the Ascomycota. Results on the number of OTUs, Chao1 index, ACE index, and Shannon index showed that Lianzhifan solution had high abundance and diversity of fungal community, and total abundance and diversity of the upper and lower layers gradually increased during the fermentation process. Among them, the abundance and diversity of the upper samples increased during the whole process, while the lower samples showed a decrease firstly and followed by a dramatically increase, despite the diversity of the upper layer was obviously higher than that of the lower layer. Conclusion Illumina HiSeq sequencing technique can be used to determine the dynamic changes of fungal communities in structure, diversity, and abundance during the process of fermentation, which provides a reference for the scientific fermentation.

四川省科学技术厅项目（2017JY0345，2015SZ0039）；四川省中医药科学技术研究专项（2016Q075）；四川省教育厅重点项目（14ZA0035）