Dataset Information

Comparison of three clustering approaches for detecting novel environmental microbial diversity.

ABSTRACT: Discovery of novel diversity in high-throughput sequencing studies is an important aspect in environmental microbial ecology. To evaluate the effects that amplicon clustering methods have on the discovery of novel diversity, we clustered an environmental marine high-throughput sequencing dataset of protist amplicons together with reference sequences from the taxonomically curated Protist Ribosomal Reference (PR(2)) database using three de novo approaches: sequence similarity networks, USEARCH, and Swarm. The potentially novel diversity uncovered by each clustering approach differed drastically in the number of operational taxonomic units (OTUs) and in the number of environmental amplicons in these novel diversity OTUs. Global pairwise alignment comparisons revealed that numerous amplicons classified as potentially novel by USEARCH and Swarm were more than 97% similar to references of PR(2). Using shortest path analyses on sequence similarity network OTUs and Swarm OTUs we found additional novel diversity within OTUs that would have gone unnoticed without further exploiting their underlying network topologies. These results demonstrate that graph theory provides powerful tools for microbial ecology and the analysis of environmental high-throughput sequencing datasets. Furthermore, sequence similarity networks were most accurate in delineating novel diversity from previously discovered diversity.

SUBMITTER: Forster D

PROVIDER: S-EPMC4782723 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Comparison of three clustering approaches for detecting novel environmental microbial diversity.

Forster Dominik D Dunthorn Micah M Stoeck Thorsten T Mahé Frédéric F

PeerJ 20160225

Discovery of novel diversity in high-throughput sequencing studies is an important aspect in environmental microbial ecology. To evaluate the effects that amplicon clustering methods have on the discovery of novel diversity, we clustered an environmental marine high-throughput sequencing dataset of protist amplicons together with reference sequences from the taxonomically curated Protist Ribosomal Reference (PR(2)) database using three de novo approaches: sequence similarity networks, USEARCH, a ...[more]

PMID: 26966652

Similar Datasets

Project description:Soil microbes act as "players" in regulating biogeochemical cycles, whereas environmental heterogeneity drives microbial community assembly patterns and is influenced by stochastic and deterministic ecological processes. Currently, the limited understanding of soil microbial community assembly patterns and interactions under temperate forest stand differences pose a challenge in studying the soil microbial involvement during the succession from coniferous to broad-leaved forests. This study investigated the changes in soil bacterial and fungal community diversity and community structure at the regional scale and identified the pathways influencing soil microbial assembly patterns and their interactions. The results showed that broad-leaved forest cover in temperate forests significantly increased soil pH, and effectively increased soil water content, total carbon (TC), total nitrogen (TN), and total phosphorus (TP) contents. Both soil bacterial and fungal alpha diversity indices were correlated with soil physicochemical properties, especially in broad-leaved forest. The bacterial and fungal community composition of coniferous forest was dominated by deterministic process (bacteria: 69.4%; fungi: 88.9%), while the bacterial community composition of broad-leaved forest was dominated by stochastic process (77.8%) and the fungal community composition was dominated by deterministic process (52.8%). Proteobacteria, Acidobacteriota, Actinobacteriota, and Verrucomicrobiota were the dominant phyla of soil bacterial communities in temperate forests. Whereas Ascomycota, Mortierellomycota, Basidiomycota, and Rozellomycota were the dominant phyla of soil fungal communities in temperate forests. Most members of dominant phylum were regulated by soil physical and chemical properties. In addition, the succession from temperate coniferous forest to broad-leaved forest was conducive to maintaining the complex network of soil bacteria and fungi, and the top 20 degree of the major taxa in the network reflected the positive response of microbial interactions to the changes of soil nutrients during forest succession. This study not only shows the mechanism by which species differences in temperate forests of northern China affect soil microbial community assembly processes, but also further emphasizes the importance of the soil microbiome as a key ecosystem factor through co-occurrence network analysis.

Project description:BackgroundEnvironmental mycobacteria (EM) include species commonly found in various terrestrial and aquatic environments, encompassing animal and human pathogens in addition to saprophytes. Approximately 150 EM species can be separated into fast and slow growers based on sequence and copy number differences of their 16S rRNA genes. Cultivation methods are not appropriate for diversity studies; few studies have investigated EM diversity in soil despite their importance as potential reservoirs of pathogens and their hypothesized role in masking or blocking M. bovis BCG vaccine.MethodsWe report here the development, optimization and validation of molecular assays targeting the 16S rRNA gene to assess diversity and prevalence of fast and slow growing EM in representative soils from semi tropical and temperate areas. New primer sets were designed also to target uniquely slow growing mycobacteria and used with PCR-DGGE, tag-encoded Titanium amplicon pyrosequencing and quantitative PCR.ResultsPCR-DGGE and pyrosequencing provided a consensus of EM diversity; for example, a high abundance of pyrosequencing reads and DGGE bands corresponded to M. moriokaense, M. colombiense and M. riyadhense. As expected pyrosequencing provided more comprehensive information; additional prevalent species included M. chlorophenolicum, M. neglectum, M. gordonae, M. aemonae. Prevalence of the total Mycobacterium genus in the soil samples ranged from 2.3×10(7) to 2.7×10(8) gene targets g(-1); slow growers prevalence from 2.9×10(5) to 1.2×10(7) cells g(-1).ConclusionsThis combined molecular approach enabled an unprecedented qualitative and quantitative assessment of EM across soil samples. Good concordance was found between methods and the bioinformatics analysis was validated by random resampling. Sequences from most pathogenic groups associated with slow growth were identified in extenso in all soils tested with a specific assay, allowing to unmask them from the Mycobacterium whole genus, in which, as minority members, they would have remained undetected.

Dataset Information

Comparison of three clustering approaches for detecting novel environmental microbial diversity.

Publications

Comparison of three clustering approaches for detecting novel environmental microbial diversity.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets