基于分级过滤的自然水体原核微生物群落分析

Natural Water Prokaryote Community Study Based on Grading Filtration

自然水体中微生物种类丰富,采集了上海周边的湖水、河水、海(岸)水、井水四类水体各两处样品,设计了含0.1μm终端截留孔径的半自动分级过滤装置分离富集不同粒径的水体微生物,使用原核微生物通用引物对16S rRNA基因V3+V4区扩增,高通量测序获得序列,对四类水体中的微生物群落进行比较分析,关注了能穿透0.22μm常规除菌过滤孔径的超微小微生物。结果显示水体环境主导了微生物的群落差异,河水与湖水中的群落总体接近,不同于海(岸)水和井水;海(岸)水和井水的微生物群落差异较大。0.1μm上截留的优势超微小微生物在河水与湖水样品中均为Proteobacteria门SAR11 clade目的CladeⅢ和Actinobacteria门Sporichthyaceae科HgcI clade,海(岸)水中为Proteobacteria门SAR11 clade目的Clade Ia和Nanoarchaeota门Woesearchaeia古菌,井水中为Nanoarchaeota门Woesearchaeia属古菌以及未定域或未分类的微生物。其中未定域的微生物16S rRNA基因在系统发育树上单独成簇,可能是古菌的一个新类型。本研究所使用的分级过滤装置显示了在发掘超微小微生物上的潜力。

Microorganisms are abundant in category in natural waters,in this study,two water samples from four categories of waters,lake,river,coast,and well water are collected from around Shanghai.A semi-auto grading filtration device with 0.1μm pore size terminal retention is designed to enrich different particle diameter microorganisms.Using universal prokaryote primer pair to amplify the V3+V4 region of 16 S rRNA gene.Prokaryote community is analyzed based on high throughput sequencing results,focusing on those ultra-small prokaryotes which physically penetrated normal sterilizing 0.22μm filter.The result showed that waters environment dominates the microbial community differences,communities in river and lake waters are close in general,however,microbial communities differ from coast and well waters,coast waters and well waters are also comparatively different from each other.The dominant ultra-small microbes retained by 0.1μm in river and lake samples are all belonged to CladeⅢof SAR11 clade order of the phylum of Proteobacteria and Sporichthyaceae family HgcI clade genus of the genus of the phylum of Actinobacteria.Clade Ia in SAR11 clade order of the phylum of Proteobacteria and archaea Woesearchaeia of the phylum of Nanoarchaeota;In well water they microorganisms belonged to archaea Woesearchaeia of the phylum of Nanoarchaeota as well as unidentified area or unclassified microorganisms.16 S rRNA gene among unclassified area microorganisms formed independent taxonomic cluster in phylogenetic tree,it may be a new archaea type.The grading filtration device used in this study showed a potential in ultra-small microorganism exploration.