Koo Hyunmin, Hakim Joseph A, Morrow Casey D, Eipers Peter G, Davila Alfonso, Andersen Dale T, Bej Asim K
Department of Biology, University of Alabama at Birmingham, Birmingham, AL, USA.
Department of Biology, University of Alabama at Birmingham, Birmingham, AL, USA.
J Microbiol Methods. 2017 Sep;140:15-22. doi: 10.1016/j.mimet.2017.06.017. Epub 2017 Jun 24.
In this study, using NextGen sequencing of the collective 16S rRNA genes obtained from two sets of samples collected from Lake Obersee, Antarctica, we compared and contrasted two bioinformatics tools, PICRUSt and Tax4Fun. We then developed an R script to assess the taxonomic and predictive functional profiles of the microbial communities within the samples. Taxa such as Pseudoxanthomonas, Planctomycetaceae, Cyanobacteria Subsection III, Nitrosomonadaceae, Leptothrix, and Rhodobacter were exclusively identified by Tax4Fun that uses SILVA database; whereas PICRUSt that uses Greengenes database uniquely identified Pirellulaceae, Gemmatimonadetes A1-B1, Pseudanabaena, Salinibacterium and Sinobacteraceae. Predictive functional profiling of the microbial communities using Tax4Fun and PICRUSt separately revealed common metabolic capabilities, while also showing specific functional IDs not shared between the two approaches. Combining these functional predictions using a customized R script revealed a more inclusive metabolic profile, such as hydrolases, oxidoreductases, transferases; enzymes involved in carbohydrate and amino acid metabolisms; and membrane transport proteins known for nutrient uptake from the surrounding environment. Our results present the first molecular-phylogenetic characterization and predictive functional profiles of the microbial mat communities in Lake Obersee, while demonstrating the efficacy of combining both the taxonomic assignment information and functional IDs using the R script created in this study for a more streamlined evaluation of predictive functional profiles of microbial communities.
在本研究中,我们利用下一代测序技术对从南极洲奥伯湖采集的两组样本中获得的16S rRNA基因集合进行分析,比较并对比了两种生物信息学工具PICRUSt和Tax4Fun。然后,我们开发了一个R脚本,以评估样本中微生物群落的分类学和预测功能概况。使用SILVA数据库的Tax4Fun专门鉴定出了诸如假黄单胞菌属、浮霉菌科、蓝藻菌纲第三亚纲、亚硝化单胞菌科、纤发菌属和红杆菌属等分类群;而使用Greengenes数据库的PICRUSt则独特地鉴定出了皮氏菌科、芽单胞菌门A1 - B1、假鱼腥藻属、盐杆菌属和中华杆菌科。分别使用Tax4Fun和PICRUSt对微生物群落进行预测功能概况分析,揭示了共同的代谢能力,同时也显示了两种方法之间未共享的特定功能ID。使用定制的R脚本合并这些功能预测,揭示了更具包容性的代谢概况,如水解酶、氧化还原酶、转移酶;参与碳水化合物和氨基酸代谢的酶;以及负责从周围环境摄取营养的膜转运蛋白。我们的研究结果首次展示了奥伯湖微生物垫群落的分子系统发育特征和预测功能概况,同时证明了结合本研究中创建的R脚本的分类学分配信息和功能ID,能够更简化地评估微生物群落的预测功能概况。