Research article - Peer-reviewed, 2020
High-Throughput Sequencing and Unsupervised Analysis of Formyltetrahydrofolate Synthetase (FTHFS) Gene Amplicons to Estimate Acetogenic Community StructureSingh, Abhijeet; Nylander, Johan A. A.; Schnurer, Anna; Bongcam-Rudloff, Erik; Mueller, Bettina
AbstractThe formyltetrahydrofolate synthetase (FTHFS) gene is a molecular marker of choice to study the diversity of acetogenic communities. However, current analyses are limited due to lack of a high-throughput sequencing approach for FTHFS gene amplicons and a dedicated bioinformatics pipeline for data analysis, including taxonomic annotation and visualization of the sequence data. In the present study, we combined the barcode approach for multiplexed sequencing with unsupervised data analysis to visualize acetogenic community structure. We used samples from a biogas digester to develop proof-of-principle for our combined approach. We successfully generated high-throughput sequence data for the partial FTHFS gene and performed unsupervised data analysis using the novel bioinformatics pipeline "AcetoScan" presented in this study, which resulted in taxonomically annotated OTUs, phylogenetic tree, abundance plots and diversity indices. The results demonstrated that high-throughput sequencing can be used to sequence the FTHFS amplicons from a pool of samples, while the analysis pipeline AcetoScan can be reliably used to process the raw sequence data and visualize acetogenic community structure. The method and analysis pipeline described in this paper can assist in the identification and quantification of known or potentially new acetogens. The AcetoScan pipeline is freely available at.
KeywordsFTHFS; acetogens; AcetoScan; AcetoBase; high-throughput sequencing
Published inFrontiers in Microbiology
2020, volume: 11, article number: 2066
Publisher: FRONTIERS MEDIA SA
Nylander, Johan A. A.
Swedish Museum of Natural History
Bongcam-Rudloff, Erik (Bongcam Rudloff, Erik)
Swedish University of Agricultural Sciences, Department of Animal Breeding and Genetics
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