<scp>MetaZooGene</scp> Intercalibration Experiment ( <scp>MZG</scp> ‐ <scp>ICE</scp> ): Metabarcoding Marine Zooplankton Diversity of the Global Ocean

DNA metabarcoding of zooplankton biodiversity is used increasingly for monitoring global ocean ecosystems, requiring comparable data from different research laboratories and ocean regions. The MetaZooGene Intercalibration Experiment (MZG-ICE) was designed to examine1 and analyse patterns of variation of DNA sequence data resulting from multi-gene metabarcoding of 10 zooplankton samples carried out by 10 research groups affiliated with the Scientific Committee for Ocean Research (SCOR). Aliquots of DNA extracted from the 10 zooplankton samples were distributed to MZG-ICE groups for metabarcoding of four gene regions: V1-V2, V4 and V9 of nuclear 18S rRNA and mitochondrial COI. Molecular protocols and procedures were recommended; substitutions were allowed as necessary. Resulting data were uploaded to a common repository for centralised statistics and bioinformatics. Based on proportional sequence numbers for abundant phyla, overall patterns of variation were consistent across many-but not all-MZG-ICE groups. V9 showed highest similarity, followed (in order) by V4, V1-V2, and COI. Outlier data were hypothesised to result from the use of different PCR protocols and sequencing platforms, and possible contamination. MZG-ICE results indicated that DNA metabarcoding data from different laboratories and research groups can provide reliable, accurate and valid descriptions of biodiversity of zooplankton throughout the ocean. Recommendations included: pre-screening QA/QC of raw data, detailed records for laboratory protocols, reagents, and instrumentation, and centralised bioinformatics and multivariate statistics. In the absence of universal agreement on standardised protocols or best practices, intercalibration is the best way forward toward validation of DNA metabarcoding of zooplankton diversity for global ocean monitoring.

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Journal
Molecular Ecology Resources
Published
2025-12-26
DOI
https://doi.org/10.1111/1755-0998.70090
Primary Topic
Environmental DNA in Biodiversity Studies
Type
article
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0.00

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article

MetaZooGene Intercalibration Experiment ( MZG ‐ ICE ): Metabarcoding Marine Zooplankton Diversity of the Global Ocean

Elza Duijm, Todd O’Brien, Rubén Escribano, Anna J. MacDonald et al.
Molecular Ecology Resources
Environmental DNA in Biodiversity Studies
article

MetaZooGene Intercalibration Experiment ( MZG ‐ ICE ): Metabarcoding Marine Zooplankton Diversity of the Global Ocean

Elza Duijm, Todd O’Brien, Rubén Escribano, Anna J. MacDonald, Agata Weydmann‐Zwolicka, Monika Mioduchowska, Jennifer M. Questel, L. Suter, Carolina E. González, Ashrenee Govender, Junya Hirai, Paola G. Batta‐Lona, Jenny A. Huggett, Andrea Polanowski, Rocio Rodriguez‐Perez, Katja T. C. A. Peijnenburg, Stacey Dubbeldam, Tone Falkenhaug, Elizaveta Ershova‐Menze, Sahar Khodami, Pedro Martinez Arbizu, Leocadio Blanco‐Bercial, Ann Bucklin, Johan Groeneveld
article en

Abstract

DNA metabarcoding of zooplankton biodiversity is used increasingly for monitoring global ocean ecosystems, requiring comparable data from different research laboratories and ocean regions. The MetaZooGene Intercalibration Experiment (MZG-ICE) was designed to examine1 and analyse patterns of variation of DNA sequence data resulting from multi-gene metabarcoding of 10 zooplankton samples carried out by 10 research groups affiliated with the Scientific Committee for Ocean Research (SCOR). Aliquots of DNA extracted from the 10 zooplankton samples were distributed to MZG-ICE groups for metabarcoding of four gene regions: V1-V2, V4 and V9 of nuclear 18S rRNA and mitochondrial COI. Molecular protocols and procedures were recommended; substitutions were allowed as necessary. Resulting data were uploaded to a common repository for centralised statistics and bioinformatics. Based on proportional sequence numbers for abundant phyla, overall patterns of variation were consistent across many-but not all-MZG-ICE groups. V9 showed highest similarity, followed (in order) by V4, V1-V2, and COI. Outlier data were hypothesised to result from the use of different PCR protocols and sequencing platforms, and possible contamination. MZG-ICE results indicated that DNA metabarcoding data from different laboratories and research groups can provide reliable, accurate and valid descriptions of biodiversity of zooplankton throughout the ocean. Recommendations included: pre-screening QA/QC of raw data, detailed records for laboratory protocols, reagents, and instrumentation, and centralised bioinformatics and multivariate statistics. In the absence of universal agreement on standardised protocols or best practices, intercalibration is the best way forward toward validation of DNA metabarcoding of zooplankton diversity for global ocean monitoring.

Molecular Ecology ResourcesVol. 26(1)
NOAA National Marine Fisheries Service (US), Naturalis Biodiversity Center (NL), Bermuda Institute of Ocean Sciences (BM), Department of Agriculture, Land Reform and Rural Development (ZA), Australian Antarctic Division (AU), University of Alaska Fairbanks (US), University of Concepción (CL), Norwegian Institute of Marine Research (NO), University of Gdańsk (PL), South African Association for Marine Biological Research (ZA), Connecticut Sea Grant (US), Millennium Institute of Oceanography (CL), Senckenberg am Meer (DE), Department of Climate Change, Energy, the Environment and Water (AU), The University of Tokyo (JP), University of Amsterdam (NL)
National Science Foundation, University of Connecticut, Norges Forskningsråd, International Council for the Exploration of the Sea, Naturalis Biodiversity Center, Instituto Milenio de Oceanografía, Agencia Nacional de Investigación y Desarrollo, National Oceanic and Atmospheric Administration, Australian Antarctic Division
Life below water
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Environmental DNA in Biodiversity Studies
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