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Investigating Microbial Eukaryotic Diversity from a Global Census: Insights from a Comparison of Pyrotag and Full-Length Sequences of 18S rRNA Genes

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Title Investigating Microbial Eukaryotic Diversity from a Global Census: Insights from a Comparison of Pyrotag and Full-Length Sequences of 18S rRNA Genes
Names Lie, Alle A. Y. (creator)
Liu, Zhenfeng (creator)
Hu, Sarah K. (creator)
Sherr, Evelyn B. (creator)
Sherr, Barry F. (creator)
et al. (creator)
Date Issued 2014-05-09 (iso8601)
Note This is the publisher’s final pdf. The published article is copyrighted by the American Society for Microbiology and can be found at: http://aem.asm.org/.
Abstract Next-generation DNA sequencing (NGS) approaches are rapidly surpassing Sanger sequencing for characterizing the diversity of
natural microbial communities. Despite this rapid transition, few comparisons exist between Sanger sequences and the generally
much shorter reads of NGS. Operational taxonomic units (OTUs) derived from full-length (Sanger sequencing) and pyrotag (454
sequencing of the V9 hypervariable region) sequences of 18S rRNA genes from 10 global samples were analyzed in order to compare
the resulting protistan community structures and species richness. Pyrotag OTUs called at 98% sequence similarity yielded
numbers of OTUs that were similar overall to those for full-length sequences when the latter were called at 97% similarity. Singleton
OTUs strongly influenced estimates of species richness but not the higher-level taxonomic composition of the community.
The pyrotag and full-length sequence data sets had slightly different taxonomic compositions of rhizarians, stramenopiles,
cryptophytes, and haptophytes, but the two data sets had similarly high compositions of alveolates. Pyrotag-based OTUs were
often derived from sequences that mapped to multiple full-length OTUs at 100% similarity. Thus, pyrotags sequenced from a
single hypervariable region might not be appropriate for establishing protistan species-level OTUs. However, nonmetric multidimensional
scaling plots constructed with the two data sets yielded similar clusters, indicating that beta diversity analysis results
were similar for the Sanger and NGS sequences. Short pyrotag sequences can provide holistic assessments of protistan communities,
although care must be taken in interpreting the results. The longer reads (>500 bp) that are now becoming available
through NGS should provide powerful tools for assessing the diversity of microbial eukaryotic assemblages.
Genre Article
Topic Operational taxonomic units
Identifier Lie, A. A. Y., Liu, Z., Hu, S. K., Jones, A. C., Kim, D. Y., Countway, P. D., ... & Caron, D. A. (2014). Investigating Microbial Eukaryotic Diversity from a Global Census: Insights from a Comparison of Pyrotag and Full-Length Sequences of 18S rRNA Genes. Applied and Environmental Microbiology, 80(14), 4363-4373. doi:10.1128/AEM.00057-14

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