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      SymPortal: A novel analytical framework and platform for coral algal symbiont next‐generation sequencing ITS2 profiling

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          Abstract

          We present SymPortal (SymPortal.org), a novel analytical framework and platform for genetically resolving the algal symbionts of reef corals using next‐generation sequencing (NGS) data of the ITS2 rDNA. Although the ITS2 marker is widely used to genetically characterize taxa within the family Symbiodiniaceae (formerly the genus Symbiodinium), the multicopy nature of the marker complicates its use. Commonly, the intragenomic diversity resultant from this multicopy nature is collapsed by analytical approaches, thereby focusing on only the most abundant sequences. In contrast, SymPortal employs logic to identify within‐sample informative intragenomic sequences, which we have termed ‘defining intragenomic variants' (DIVs), to identify ITS2‐type profiles representative of putative Symbiodiniaceae taxa. By making use of this intragenomic ITS2 diversity, SymPortal is able to resolve genetic delineations using the ITS2 marker at a level that was previously only possible by using additional genetic markers. We demonstrate this by comparing this novel approach to the most commonly used alternative approach for NGS ITS2 data, the 97% similarity clustering to operational taxonomic units (OTUs). The SymPortal platform accepts NGS raw sequencing data as input to provide an easy‐to‐use, standardization‐enforced, and community‐driven framework that integrates with a database to gain resolving power with increased use. We consider that SymPortal, in conjunction with ongoing large‐scale sampling and sequencing efforts, should play an instrumental role in making future sampling efforts more comparable and in maximizing their efficacy in working towards the classification of the global Symbiodiniaceae diversity.

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          Landscape ecology of algal symbionts creates variation in episodes of coral bleaching.

          Reef-building corals are obligate, mutualistic symbioses of heterotrophic animals and phototrophic dinoflagellates (Symbiodinium spp.). Contrary to the earlier, widely accepted belief that corals harbour only one symbiont, we found that the ecologically dominant Caribbean corals Montastraea annularis and M. faveolata can act as hosts to dynamic, multi-species communities of Symbiodinium. Composition of these communities follows gradients of environmental irradiance, implying that physiological acclimatization is not the only mechanism by which corals cope with environmental heterogeneity. The importance of this diversity was underlined by analysis of a natural episode of coral bleaching. Patterns of bleaching could be explained by the preferential elimination of a symbiont associated with low irradiance from the brightest parts of its distribution. Comparative analyses of symbionts before and after bleaching from the same corals supported this interpretation, and suggested that some corals were protected from bleaching by hosting an additional symbiont that is more tolerant of high irradiance and temperature. This 'natural experiment' suggests that temporal and spatial variability can favour the coexistence of diverse symbionts within a host, despite the potential for destabilizing competition among them.
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            Long-standing environmental conditions, geographic isolation and host-symbiont specificity influence the relative ecological dominance and genetic diversification of coral endosymbionts in the genusSymbiodinium

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              Change in algal symbiont communities after bleaching, not prior heat exposure, increases heat tolerance of reef corals.

              Mutualistic organisms can be particularly susceptible to climate change stress, as their survivorship is often limited by the most vulnerable partner. However, symbiotic plasticity can also help organisms in changing environments by expanding their realized niche space. Coral-algal (Symbiodinium spp.) symbiosis exemplifies this dichotomy: the partnership is highly susceptible to 'bleaching' (stress-induced symbiosis breakdown), but stress-tolerant symbionts can also sometimes mitigate bleaching. Here, we investigate the role of diverse and mutable symbiotic partnerships in increasing corals' ability to thrive in high temperature conditions. We conducted repeat bleaching and recovery experiments on the coral Montastraea cavernosa, and used quantitative PCR and chlorophyll fluorometry to assess the structure and function of Symbiodinium communities within coral hosts. During an initial heat exposure (32 °C for 10 days), corals hosting only stress-sensitive symbionts (Symbiodinium C3) bleached, but recovered (at either 24 °C or 29 °C) with predominantly (>90%) stress-tolerant symbionts (Symbiodinium D1a), which were not detected before bleaching (either due to absence or extreme low abundance). When a second heat stress (also 32 °C for 10 days) was applied 3 months later, corals that previously bleached and were now dominated by D1a Symbiodinium experienced less photodamage and symbiont loss compared to control corals that had not been previously bleached, and were therefore still dominated by Symbiodinium C3. Additional corals that were initially bleached without heat by a herbicide (DCMU, at 24 °C) also recovered predominantly with D1a symbionts, and similarly lost fewer symbionts during subsequent thermal stress. Increased thermotolerance was also not observed in C3-dominated corals that were acclimated for 3 months to warmer temperatures (29 °C) before heat stress. These findings indicate that increased thermotolerance post-bleaching resulted from symbiont community composition changes, not prior heat exposure. Moreover, initially undetectable D1a symbionts became dominant only after bleaching, and were critical to corals' resilience after stress and resistance to future stress.
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                Author and article information

                Contributors
                benjamin.hume@kaust.edu.sa
                christian.voolstra@kaust.edu.sa
                Journal
                Mol Ecol Resour
                Mol Ecol Resour
                10.1111/(ISSN)1755-0998
                MEN
                Molecular Ecology Resources
                John Wiley and Sons Inc. (Hoboken )
                1755-098X
                1755-0998
                26 April 2019
                July 2019
                : 19
                : 4 ( doiID: 10.1111/men.2019.19.issue-4 )
                : 1063-1080
                Affiliations
                [ 1 ] Division of Biological and Environmental Science and Engineering (BESE), Red Sea Research Center King Abdullah University of Science and Technology (KAUST) Thuwal Saudi Arabia
                [ 2 ] Center for Genomics and Systems Biology New York University Abu Dhabi Abu Dhabi UAE
                [ 3 ] Faculty of Computer Science and Technology University of Cambridge UK
                [ 4 ] Department of Biology The Pennsylvania State University University Park Pennsylvania
                [ 5 ] Coral Reef Laboratory, Ocean and Earth Sciences University of Southampton Southampton UK
                [ 6 ] Institute for Life Sciences University of Southampton Southampton UK
                Author notes
                [*] [* ] Correspondence

                Benjamin C. C. Hume and Christian R. Voolstra, Red Sea Research Center, Division of Biological and Environmental Science and Engineering (BESE), King Abdullah University of Science and Technology (KAUST), Thuwal, Saudi Arabia.

                Emails: benjamin.hume@ 123456kaust.edu.sa ; christian.voolstra@ 123456kaust.edu.sa

                Author information
                https://orcid.org/0000-0001-7753-3903
                https://orcid.org/0000-0003-2237-9261
                https://orcid.org/0000-0001-6087-6424
                https://orcid.org/0000-0003-4555-3795
                Article
                MEN13004
                10.1111/1755-0998.13004
                6618109
                30740899
                6d2c779e-f7d7-4697-a816-cc15ff2854dc
                © 2019 The Authors. Molecular Ecology Resources Published by 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.

                History
                : 07 October 2018
                : 30 January 2019
                : 31 January 2019
                Page count
                Figures: 5, Tables: 3, Pages: 18, Words: 13749
                Funding
                Funded by: Natural Environment Research Council
                Award ID: NE/K00641X/1
                Funded by: King Abdullah University of Science and Technology (KAUST)
                Award ID: Baseline funding
                Funded by: FP7 Ideas: European Research Council
                Award ID: FP7/2007-013/ERC 311179
                Categories
                Resource Article
                RESOURCE ARTICLES
                Computer Programs
                Custom metadata
                2.0
                men13004
                July 2019
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.6.5 mode:remove_FC converted:10.07.2019

                Ecology
                its2,molecular ecology,multicopy,phylogentics,symbiodiniaceae,symportal
                Ecology
                its2, molecular ecology, multicopy, phylogentics, symbiodiniaceae, symportal

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