Combining artificial substrates, morphology and DNA metabarcoding for investigating macrozoobenthic communities in NW Iberia
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1
Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Portugal
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2
Institute of Science and Innovation for Bio-Sustainability, University of Minho, Portugal
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3
Estación de Ciencias Marinas de Toralla (ECIMAT), Spain
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4
Department of Ecology and Animal Biology, University of Vigo, Spain
Problem statement: Assessment of species composition in zoobenthic communities is a key element of biomonitoring in marine waters. However, marine biodiversity assessment in large-scale biomonitoring programs is challenging because it is commonly based on morphological identifications, which are time-consuming and require taxonomic specialists. The emergence of a new generation of technological DNA-based tools is transforming our ability to study patterns and trends of biodiversity. DNA metabarcoding provides the opportunity to improve accuracy and throughput compared to morphology-based assessments. Although some methodological optimization is still required, high-throughput techniques coupled with standardized sampling strategies accurately improve the ability to monitor marine communities with unprecedented potential for precision and detail. Different artificial substrates deployed in coastal areas may selectively influence and enhance the colonization of different species determining the resultant community. The creation of permanent monitoring stations using artificial substrates provides an alternative to easily and routinely assess biodiversity. Objective: Here, we test the influence of different types of artificial substrates on the composition and structure of marine macrozoobenthic communities. In order to achieve this goal, we combined morphology and DNA metabarcoding approaches to monitor coastal macrozoobenthic communities and investigate the short-term and seasonal patterns of colonization in artificial substrates. Methodology: Three different types of artificial substrates were used: slate, polyvinyl chloride (PVC) and granite. In December 2016, 16 replicates of each substrate were randomly deployed close to the dock of Toralla Island (Vigo, NW Spain). After 3, 7, 10 and 15 months, four replicates of each substrate were randomly removed. Three replicates of each substrate were used for morphology and one replicate of each substrate was used for DNA metabarcoding. The mobile and sessile fauna were separated and preserved for subsequent analysis. DNA amplification was performed for an internal region of the COI barcode (~313 bp) and for the V4 region from 18S rRNA gene (ca. 400 bp). Results: Compared to morphology, DNA metabarcoding retrieved more taxa and higher taxonomic resolution. However, some of the species identified with morphology were not detected using DNA metabarcoding (e.g. Pecten maximus and Halocynthia papillosa). While sampling strategy, samples processing or biomass can explain some of these cases, attention should be paid for the non-detection with DNA of very representative species, like the balanus Perforatus perforatus. Compared to the other substrates, slate replicates resulted in a higher colonization of target taxa and granite replicates have the lower rate of species detected. Some taxa revealed different time-patterns to colonize the substrates (e.g. Ampithoe rubricata and Hiatella arctica) as well as variations in taxonomic diversity among substrates and seasons. While some species were evenly detected in all/nearly all of the substrates and time combinations (e.g. Obelia geniculata/Jassa slatteryi), some taxon were time-specific (e.g. Limacia clavigera only detected in second recruitment time – after 7 months; Dexamine spinosa detected in all substrates but only in third recruitment time - after 10 months) or randomly detected with variations in taxonomic diversity among substrates and seasons (e.g. Myrianida prolifera). The adopted approaches also demonstrated to be efficient to detect non-indigenous species (e.g. Perinereis aibuhitensis: a commercially very important polychaete also used for biomonitoring studies). Major conclusions: In general, artificial substrates promote macrozoobenthic colonization and coupled with molecular tools capture the composition of natural community. This highlights the importance of the adopted sampling strategy to be used in biomonitoring programs to capture benthic biodiversity. The results also illustrate the influence of substrate and season in the recruitment of zoobenthos. Although DNA metabarcoding detected a higher number of taxa and with more resolution, the two methods applied were somewhat complementary in their ability to detect benthic species and both should be used to avoid missing relevant taxa. This project provided an opportunity to yields insights for a better comprehension of fundamental ecological processes in macrozoobenthic communities and opening new perspectives to be used in other studies to supply more extensive, detailed and rigorous data for marine resource management and biodiversity conservation in coastal ecosystems.
Acknowledgements
This study was supported by the project The NextSea (NORTE-01-0145-FEDER-000032), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). BL benefitted from an FCT fellowship PD/BD/127994/2016.
Keywords:
DNA metabarcoding,
High-throughput sequencing (HTS),
biomonitoring,
Artificial substrates,
Marine macroinvertebrates
Conference:
XX Iberian Symposium on Marine Biology Studies (SIEBM XX) , Braga, Portugal, 9 Sep - 12 Sep, 2019.
Presentation Type:
Oral Presentation
Topic:
Ecology, Biodiversity and Vulnerable Ecosystems
Citation:
Leite
BR,
Vieira
PE,
Troncoso
JS and
Costa
FO
(2019). Combining artificial substrates, morphology and DNA metabarcoding for investigating macrozoobenthic communities in NW Iberia.
Front. Mar. Sci.
Conference Abstract:
XX Iberian Symposium on Marine Biology Studies (SIEBM XX) .
doi: 10.3389/conf.fmars.2019.08.00061
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Received:
23 May 2019;
Published Online:
27 Sep 2019.
*
Correspondence:
MD. Barbara R Leite, Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga, Portugal, barbaradrl.bio@gmail.com