Open Access Repository

Effect of marker choice and thermal cycling protocol on zooplankton DNA metabarcoding studies

Downloads

Downloads per month over past year

Clarke, LJ ORCID: 0000-0002-0844-4453, Beard, JM, Swadling, KM ORCID: 0000-0002-7620-841X and Deagle, BE 2017 , 'Effect of marker choice and thermal cycling protocol on zooplankton DNA metabarcoding studies' , Ecology and Evolution, vol. 7, no. 3 , pp. 873-883 , doi: 10.1002/ece3.2667.

[img]
Preview
PDF
114582 final.pdf | Download (699kB)

| Preview

Abstract

DNA metabarcoding is a promising approach for rapidly surveying biodiversity and is likely to become an important tool for measuring ecosystem responses to environmental change. Metabarcoding markers need sufficient taxonomic coverage to detect groups of interest, sufficient sequence divergence to resolve species, and will ideally indicate relative abundance of taxa present. We characterized zooplankton assemblages with three different metabarcoding markers (nuclear 18S rDNA, mitochondrial COI, and mitochondrial 16S rDNA) to compare their performance in terms of taxonomic coverage, taxonomic resolution, and correspondence between morphology- and DNA-based identification. COI amplicons sequenced on separate runs showed that operational taxonomic units representing >0.1% of reads per sample were highly reproducible, although slightly more taxa were detected using a lower annealing temperature. Mitochondrial COI and nuclear 18S showed similar taxonomic coverage across zooplankton phyla. However, mitochondrial COI resolved up to threefold more taxa to species compared to 18S. All markers revealed similar patterns of beta-diversity, although different taxa were identified as the greatest contributors to these patterns for 18S. For calanoid copepod families, all markers displayed a positive relationship between biomass and sequence reads, although the relationship was typically strongest for 18S. The use of COI for metabarcoding has been questioned due to lack of conserved primer-binding sites. However, our results show the taxonomic coverage and resolution provided by degenerate COI primers, combined with a comparatively well-developed reference sequence database, make them valuable metabarcoding markers for biodiversity assessment.

Item Type: Article
Authors/Creators:Clarke, LJ and Beard, JM and Swadling, KM and Deagle, BE
Keywords: cytochrome oxidase subunit I, environmental DNA, metabarcoding, mitochondrial 16S ribosomal DNA, nuclear 18S rDNA, zooplankton
Journal or Publication Title: Ecology and Evolution
Publisher: John Wiley & Sons Ltd.
ISSN: 2045-7758
DOI / ID Number: 10.1002/ece3.2667
Copyright Information:

Copyright 2017 The Authors. Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) https://creativecommons.org/licenses/by/4.0/

Related URLs:
Item Statistics: View statistics for this item

Actions (login required)

Item Control Page Item Control Page
TOP