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      Metabarcoding Insights Into the Trophic Behavior and Identity of Intertidal Benthic Foraminifera

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          Abstract

          Foraminifera are ubiquitous marine protists with an important role in the benthic carbon cycle. However, morphological observations often fail to resolve their exact taxonomic placement and there is a lack of field studies on their particular trophic preferences. Here, we propose the application of metabarcoding as a tool for the elucidation of the in situ feeding behavior of benthic foraminifera, while also allowing the correct taxonomic assignment of the feeder, using the V9 region of the 18S (small subunit; SSU) rRNA gene. Living foraminiferal specimens were collected from two intertidal mudflats of the Wadden Sea and DNA was extracted from foraminiferal individuals and from the surrounding sediments. Molecular analysis allowed us to confirm that our foraminiferal specimens belong to three genetic types: Ammonia sp. T6, Elphidium sp. S5 and Haynesina sp. S16. Foraminiferal intracellular eukaryote communities reflected to an extent those of the surrounding sediments but at different relative abundances. Unlike sediment eukaryote communities, which were largely determined by the sampling site, foraminiferal intracellular eukaryote communities were driven by foraminiferal species, followed by sediment depth. Our data suggests that Ammonia sp. T6 can predate on metazoan classes, whereas Elphidium sp. S5 and Haynesina sp. S16 are more likely to ingest diatoms. These observations, alongside the use of metabarcoding in similar ecological studies, significantly contribute to our overall understanding of the ecological roles of these protists in intertidal benthic environments and their position and function in the benthic food webs.

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            A Method for Studying Protistan Diversity Using Massively Parallel Sequencing of V9 Hypervariable Regions of Small-Subunit Ribosomal RNA Genes

            Background Massively parallel pyrosequencing of amplicons from the V6 hypervariable regions of small-subunit (SSU) ribosomal RNA (rRNA) genes is commonly used to assess diversity and richness in bacterial and archaeal populations. Recent advances in pyrosequencing technology provide read lengths of up to 240 nucleotides. Amplicon pyrosequencing can now be applied to longer variable regions of the SSU rRNA gene including the V9 region in eukaryotes. Methodology/Principal Findings We present a protocol for the amplicon pyrosequencing of V9 regions for eukaryotic environmental samples for biodiversity inventories and species richness estimation. The International Census of Marine Microbes (ICoMM) and the Microbial Inventory Research Across Diverse Aquatic Long Term Ecological Research Sites (MIRADA-LTERs) projects are already employing this protocol for tag sequencing of eukaryotic samples in a wide diversity of both marine and freshwater environments. Conclusions/Significance Massively parallel pyrosequencing of eukaryotic V9 hypervariable regions of SSU rRNA genes provides a means of estimating species richness from deeply-sampled populations and for discovering novel species from the environment.
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              A new Symbiodinium clade (Dinophyceae) from soritid foraminifera in Hawai'i.

              Dinoflagellates in the genus Symbiodinium are crucial components of coral reef ecosystems in their roles as endosymbionts of corals and other marine invertebrates. The genus Symbiodinium encompasses eight lineages (clades A-H), and multiple sub-clade types. Symbiodinium in clades A, B, C, and D are most commonly associated with metazoan hosts while clades C, D, F, G, and H with large soritid foraminifera. Recent studies have described a diversity of new Symbiodinium types within each clades, but no new clades have been reported since 2001. Here, we describe a new clade of Symbiodinium isolated from soritid foraminifera from Hawai'i. Published by Elsevier Inc.
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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                28 May 2019
                2019
                : 10
                : 1169
                Affiliations
                [1] 1Aquatic Biogeochemistry Research Unit, Ecosystems and Environment Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki , Helsinki, Finland
                [2] 2Biological and Environmental Sciences, University of Stirling , Stirling, United Kingdom
                [3] 3Department of Ocean Systems, NIOZ-Royal Netherlands Institute for Sea Research and Utrecht University , Den Burg, Netherlands
                Author notes

                Edited by: Ramiro Logares, Institute of Marine Sciences (ICM), Spain

                Reviewed by: Franck Lejzerowicz, University of California, San Diego, United States; Russel J. S. Orr, University of Oslo, Norway

                *Correspondence: Panagiota-Myrsini Chronopoulou, myrtuscom@ 123456gmail.com

                This article was submitted to Aquatic Microbiology, a section of the journal Frontiers in Microbiology

                Article
                10.3389/fmicb.2019.01169
                6547873
                31191490
                1dc0ec23-bd57-4552-9dfd-ca0e66899b8d
                Copyright © 2019 Chronopoulou, Salonen, Bird, Reichart and Koho.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 03 December 2018
                : 07 May 2019
                Page count
                Figures: 5, Tables: 1, Equations: 0, References: 102, Pages: 16, Words: 0
                Funding
                Funded by: Academy of Finland 10.13039/501100002341
                Award ID: 278827
                Award ID: 283453
                Award ID: 312495
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                metabarcoding,benthic foraminifera,trophic strategy,benthic food web,benthic microbial ecology,molecular phylogeny

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