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Newly designed 16S rRNA metabarcoding primers amplify diverse and novel archaeal taxa from the environment

High‐throughput studies of microbial communities suggest that Archaea are a widespread component of microbial diversity in various ecosystems. However, proper quantification of archaeal diversity and community ecology remains limited, as sequence coverage of Archaea is usually low owing to the inabi...

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Autores principales: Bahram, Mohammad, Anslan, Sten, Hildebrand, Falk, Bork, Peer, Tedersoo, Leho
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6618113/
https://www.ncbi.nlm.nih.gov/pubmed/30058291
http://dx.doi.org/10.1111/1758-2229.12684
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author Bahram, Mohammad
Anslan, Sten
Hildebrand, Falk
Bork, Peer
Tedersoo, Leho
author_facet Bahram, Mohammad
Anslan, Sten
Hildebrand, Falk
Bork, Peer
Tedersoo, Leho
author_sort Bahram, Mohammad
collection PubMed
description High‐throughput studies of microbial communities suggest that Archaea are a widespread component of microbial diversity in various ecosystems. However, proper quantification of archaeal diversity and community ecology remains limited, as sequence coverage of Archaea is usually low owing to the inability of available prokaryotic primers to efficiently amplify archaeal compared to bacterial rRNA genes. To improve identification and quantification of Archaea, we designed and validated the utility of several primer pairs to efficiently amplify archaeal 16S rRNA genes based on up‐to‐date reference genes. We demonstrate that several of these primer pairs amplify phylogenetically diverse Archaea with high sequencing coverage, outperforming commonly used primers. Based on comparing the resulting long 16S rRNA gene fragments with public databases from all habitats, we found several novel family‐ to phylum‐level archaeal taxa from topsoil and surface water. Our results suggest that archaeal diversity has been largely overlooked due to the limitations of available primers, and that improved primer pairs enable to estimate archaeal diversity more accurately.
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spelling pubmed-66181132019-07-22 Newly designed 16S rRNA metabarcoding primers amplify diverse and novel archaeal taxa from the environment Bahram, Mohammad Anslan, Sten Hildebrand, Falk Bork, Peer Tedersoo, Leho Environ Microbiol Rep Brief Reports High‐throughput studies of microbial communities suggest that Archaea are a widespread component of microbial diversity in various ecosystems. However, proper quantification of archaeal diversity and community ecology remains limited, as sequence coverage of Archaea is usually low owing to the inability of available prokaryotic primers to efficiently amplify archaeal compared to bacterial rRNA genes. To improve identification and quantification of Archaea, we designed and validated the utility of several primer pairs to efficiently amplify archaeal 16S rRNA genes based on up‐to‐date reference genes. We demonstrate that several of these primer pairs amplify phylogenetically diverse Archaea with high sequencing coverage, outperforming commonly used primers. Based on comparing the resulting long 16S rRNA gene fragments with public databases from all habitats, we found several novel family‐ to phylum‐level archaeal taxa from topsoil and surface water. Our results suggest that archaeal diversity has been largely overlooked due to the limitations of available primers, and that improved primer pairs enable to estimate archaeal diversity more accurately. John Wiley & Sons, Inc. 2018-09-12 2019-08 /pmc/articles/PMC6618113/ /pubmed/30058291 http://dx.doi.org/10.1111/1758-2229.12684 Text en © 2018 The Authors. Environmental Microbiology Reports published by Society for Applied Microbiology and John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Brief Reports
Bahram, Mohammad
Anslan, Sten
Hildebrand, Falk
Bork, Peer
Tedersoo, Leho
Newly designed 16S rRNA metabarcoding primers amplify diverse and novel archaeal taxa from the environment
title Newly designed 16S rRNA metabarcoding primers amplify diverse and novel archaeal taxa from the environment
title_full Newly designed 16S rRNA metabarcoding primers amplify diverse and novel archaeal taxa from the environment
title_fullStr Newly designed 16S rRNA metabarcoding primers amplify diverse and novel archaeal taxa from the environment
title_full_unstemmed Newly designed 16S rRNA metabarcoding primers amplify diverse and novel archaeal taxa from the environment
title_short Newly designed 16S rRNA metabarcoding primers amplify diverse and novel archaeal taxa from the environment
title_sort newly designed 16s rrna metabarcoding primers amplify diverse and novel archaeal taxa from the environment
topic Brief Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6618113/
https://www.ncbi.nlm.nih.gov/pubmed/30058291
http://dx.doi.org/10.1111/1758-2229.12684
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