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Towards an in-depth characterization of Symbiodiniaceae in tropical giant clams via metabarcoding of pooled multi-gene amplicons.

ABSTRACT: High-throughput sequencing is revolutionizing our ability to comprehensively characterize free-living and symbiotic Symbiodiniaceae, a diverse dinoflagellate group that plays a critical role in coral reef ecosystems. Most studies however, focus on a single marker for metabarcoding Symbiodiniaceae, potentially missing important ecological traits that a combination of markers may capture. In this proof-of-concept study, we used a small set of symbiotic giant clam (Tridacna maxima) samples obtained from nine French Polynesian locations and tested a dual-index sequence library preparation method that pools and simultaneously sequences multiple Symbiodiniaceae gene amplicons per sample for in-depth biodiversity assessments. The rationale for this approach was to allow the metabarcoding of multiple genes without extra costs associated with additional single amplicon dual indexing and library preparations. Our results showed that the technique effectively recovered very similar proportions of sequence reads and dominant Symbiodiniaceae clades among the three pooled gene amplicons investigated per sample, and captured varying levels of phylogenetic resolution enabling a more comprehensive assessment of the diversity present. The pooled Symbiodiniaceae multi-gene metabarcoding approach described here is readily scalable, offering considerable analytical cost savings while providing sufficient phylogenetic information and sequence coverage.

SUBMITTER: Pochon X

PROVIDER: S-EPMC6521813 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:A surprising recent discovery revealed that the brightly reflective cells ('iridocytes') in the epithelia of giant clams actually send the majority of incident photons 'forward' into the tissue. While the intracellular Bragg reflectors in these cells are responsible for their colourful back reflection, Mie scattering produces the forward scattering, thus illuminating a dense population of endosymbiotic, photosynthetic microalgae. We now present a detailed micro-spectrophotometric characterization of the Bragg stacks in the iridocytes in live tissue to obtain the refractive index of the high-index layers (1.39 to 1.58, average 1.44 ± 0.04), the thicknesses of the high- and low-index layers (50-150 nm), and the numbers of pairs of layers (2-11) that participate in the observed spectral reflection. Based on these measurements, we performed electromagnetic simulations to better understand the optical behaviour of the iridocytes. The results open a deeper understanding of the optical behaviour of these cells, with the counterintuitive discovery that specific combinations of iridocyte diameter and Bragg-lamellar spacing can produce back reflection of the same colour that is also scattered forward, in preference to other wavelengths that are scattered at higher angles. We find for all values of size and wavelength investigated that more than 90% of the incident energy is carried by the photons that are scattered in the forward direction; while this forward scattering from each iridocyte shows very narrow angular dispersion (ca ±6°), the multiplicative scattering from a layer of ca 20 iridocytes broadens this dispersion to a cone of approximately ±90°. This understanding of the complex biophotonic dynamics enhances our comprehension of the physiologically, ecologically and evolutionarily significant light environment inside the giant clam, which is diffuse and nearly white at small tissue depths and downwelling, relatively monochromatic, and can be the same colour as the back-reflected light at greater depths in the tissue. Originally thought to be unique, cells of similar structure and photonic activity are now recognized in other species, where they serve other functions. The behaviour of the iridocytes opens possible new considerations for conservation and management of the valuable giant clam resource and new avenues for biologically inspired photonic applications.

Project description:BackgroundDinoflagellates of family Symbiodiniaceae are important to coral reef ecosystems because of their contribution to coral health and growth; however, only a few studies have investigated the function and distribution of Symbiodiniaceae in Indonesia. Understanding the distribution of different kinds of Symbiodiniaceae can improve forecasting of future responses of various coral reef systems to climate change. This study aimed to determine the diversity of Symbiodiniaceae around Lombok using environmental DNA (eDNA).MethodsSeawater and sediment samples were collected from 18 locations and filtered to obtain fractions of 0.4-12 and >12 µm. After extraction, molecular barcoding polymerase chain reaction was conducted to amplify the primary V9-SSU 18S rRNA gene, followed by sequencing (Illumina MiSeq). BLAST, Naïve-fit-Bayes, and maximum likelihood routines were used for classification and phylogenetic reconstruction. We compared results across sampling sites, sample types (seawater/sediment), and filter pore sizes (fraction).ResultsPhylogenetic analyses resolved the amplicon sequence variants into 16 subclades comprising six Symbiodiniaceae genera (or genera-equivalent clades) as follows: Symbiodinium, Breviolum, Cladocopium, Durusdinium, Foraminifera Clade G, and Halluxium. Comparative analyses showed that the three distinct lineages within Cladocopium, Durusdinium, and Foraminifera Clade G were the most common. Most of the recovered sequences appeared to be distinctive of different sampling locations, supporting the possibility that eDNA may resolve regional and local differences among Symbiodiniaceae genera and species.ConclusionseDNA surveys offer a rapid proxy for evaluating Symbiodiniaceae species on coral reefs and are a potentially useful approach to revealing diversity and relative ecological dominance of certain Symbiodiniaceae organisms. Moreover, Symbiodiniaceae eDNA analysis shows potential in monitoring the local and regional stability of coral-algal mutualisms.

Project description:BACKGROUND:Giant clams and scleractinian (reef-building) corals are keystone species of coral reef ecosystems. The basis of their ecological success is a complex and fine-tuned symbiotic relationship with microbes. While the effect of environmental change on the composition of the coral microbiome has been heavily studied, we know very little about the composition and sensitivity of the microbiome associated with clams. Here, we explore the influence of increasing temperature on the microbial community (bacteria and dinoflagellates from the family Symbiodiniaceae) harbored by giant clams, maintained either in isolation or exposed to other reef species. We created artificial benthic assemblages using two coral species (Pocillopora damicornis and Acropora cytherea) and one giant clam species (Tridacna maxima) and studied the microbial community in the latter using metagenomics. RESULTS:Our results led to three major conclusions. First, the health status of giant clams depended on the composition of the benthic species assemblages. Second, we discovered distinct microbiotypes in the studied T. maxima population, one of which was disproportionately dominated by Vibrionaceae and directly linked to clam mortality. Third, neither the increase in water temperature nor the composition of the benthic assemblage had a significant effect on the composition of the Symbiodiniaceae and bacterial communities of T. maxima. CONCLUSIONS:Altogether, our results suggest that at least three microbiotypes naturally exist in the studied clam populations, regardless of water temperature. These microbiotypes plausibly provide similar functions to the clam host via alternate molecular pathways as well as microbiotype-specific functions. This redundancy in functions among microbiotypes together with their specificities provides hope that giant clam populations can tolerate some levels of environmental variation such as increased temperature. Importantly, the composition of the benthic assemblage could make clams susceptible to infections by Vibrionaceae, especially when water temperature increases. Video abstract.

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Towards an in-depth characterization of Symbiodiniaceae in tropical giant clams via metabarcoding of pooled multi-gene amplicons.

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