基于高通量测序研究厚朴“发汗”过程中微生物群落多样性及其特征

Microbial community diversity and its characteristics in Magnolia Officinalis Cortex "sweating" process based on high-throughput sequencing

厚朴为常用中药,"发汗"是厚朴药材初加工的关键步骤,也是药材质量上乘的重要体现。为探究厚朴药材"发汗"过程中的微生物群落变化特征及其在整个"发汗"过程中的变化规律,该研究采用高通量测序技术对厚朴"发汗"过程中的微生物多样性进行研究。13个送试样品共检出334个真菌菌属,674个细菌菌属。"发汗"初期,优势菌属不很明显。"发汗"中期优势真菌为假丝酵母属Candida(45.01%~71.93%),"发汗"后期以曲霉属Aspergillus(45.83%~95.51%)占绝对优势;细菌在"发汗"中后期优势菌属主要为肠杆菌属Enterobacter和克雷伯菌属Klebsiella,二者此消彼长,总比例≥56.05%。同时,芽胞杆菌属Bacillus、异常球菌属Deinococcus、鞘氨醇单胞菌属Sphingomonas、薄层菌属Hymenobacter、Jatrophihabitans等亦为优势菌属。首次开展厚朴"发汗"过程中微生物多样性研究,发现"发汗"后其"气味浓厚,内表面变为棕褐色"这一性状特征的形成可能与曲霉属Aspergillus和假丝酵母菌属Candida等微生物群落代谢有关,为进一步研究"发汗"过程中微生物对厚朴药材质量的影响提供理论依据。

Magnolia Officinalis Cortex has been used as a traditional Chinese herb for thousands of years in China.According to Chinese Pharmacopoeia,the processing of Magnolia Officinalis Cortex needs"sweating"or"Fahan",which was a special drying process and considered to be an important symbol for high quality and genuine medicinal materials.In this unique processing mode,Magnolia Officinalis Cortex’s microbial community structure may be changed,but little is known about microbial diversity during the"sweating".In this study,to analyze the change and its change rules of microbial community of Magnolia Officinalis Cortex in the whole process of"sweating",and find out the microbial community that affects the quality of Magnolia Officinalis Cortex in the process of its"sweating",and provide a basis for further research on the microbial transformation of Magnolia Officinalis Cortex,MiSeq highthroughput sequencing was used to evaluate the microbial diversity of natural"sweating"of Magnolia Officinalis Cortex.In this research,334 genera fungi and 674 genera bacteria were identified.The dominant species weren’t obvious during the early stage of"sweating".Candida was the dominant fungal species(45.01%-71.93%)during the medium"sweating"stage.Aspergillus is the dominant fungal species(45.83%-95.51%)during the late stage of"sweating".Moreover,Enterobacter and Klebsiella were the primary bacterial genus(≥56.05%)during the middle and late stages of"sweating".In addition,the predominant bacteria in the process of"sweating"included Bacillus,Deinococcus,Sphingomonas,Hymenobacter and Jatrophihabitans.In conclusion,the microbial diversities and the main dominant fungi and bacteria in the process of"sweating"of Magnolia Officinalis Cortex were initially determined.It was also found that the metabolism of Aspergillus and Candida may be related to the character formation,which were sweet odor and brown inner surface after"sweating".The results provide a theoretical basis for the study of the influence of different microorganisms on the excellent traits formation of"sweating"Magnolia Officinalis Cortex.