Symbiodinium Types Research Articles

Searching for phylogenetic patterns of Symbiodiniaceae community structure among Indo-Pacific Merulinidae corals.

Over half of all extant stony corals (Cnidaria: Anthozoa: Scleractinia) harbour endosymbiotic dinoflagellates of the family Symbiodiniaceae, forming the foundational species of modern shallow reefs. However, whether these associations are conserved on the coral phylogeny remains unknown. Here we aim to characterise Symbiodiniaceae communities in eight closely-related species in the genera Merulina, Goniastrea and Scapophyllia, and determine if the variation in endosymbiont community structure can be explained by the phylogenetic relatedness among hosts. We perform DNA metabarcoding of the nuclear internal transcribed spacer 2 using Symbiodiniaceae-specific primers on 30 coral colonies to recover three major endosymbiont clades represented by 23 distinct types. In agreement with previous studies on Southeast Asian corals, we find an abundance of Cladocopium and Durusdinium, but also detect Symbiodinium types in three of the eight coral host species. Interestingly, differences in endosymbiont community structure are dominated by host variation at the intraspecific level, rather than interspecific, intergeneric or among-clade levels, indicating a lack of phylogenetic constraint in the coral-endosymbiont association among host species. Furthermore, the limited geographic sampling of four localities spanning the Western and Central Indo-Pacific preliminarily hints at large-scale spatial structuring of Symbiodiniaceae communities. More extensive collections of corals from various regions and environments will help us better understand the specificity of the coral-endosymbiont relationship.

Ecophysiology of mesophotic reef‐building corals in Hawai‘i is influenced by symbiont–host associations, photoacclimatization, trophic plasticity, and adaptation

AbstractMesophotic reef corals remain largely unexplored in terms of the genetic adaptations and physiological mechanisms to acquire, allocate, and use energy for survival and reproduction. In the Hawaiian Archipelago, the Leptoseris species complex form the most spatially extensive mesophotic coral ecosystem known and provide habitat for a unique community. To study how the ecophysiology of Leptoseris species relates to symbiont–host specialization and understand the mechanisms responsible for coral energy acquisition in extreme low light environments, we examined Symbiodinium (endosymbiotic dinoflagellate) photobiological characteristics and the lipids and isotopic signatures from Symbiodinium and coral hosts over a depth‐dependent light gradient (55–7 μmol photons m−2 s−1, 60–132 m). Clear performance differences demonstrate different photoadaptation and photoacclimatization across this genus. Our results also show that flexibility in photoacclimatization depends primarily on Symbiodinium type. Colonies harboring Symbiodinium sp. COI‐2 showed significant increases in photosynthetic pigment content with increasing depth, whereas colonies harboring Symbiodinium spp. COI‐1 and COI‐3 showed variability in pigment composition, yield measurements for photosystem II, as well as size and density of Symbiodinium cells. Despite remarkable differences in photosynthetic adaptive strategies, there were no significant differences among lipids of Leptoseris species with depth. Finally, isotopic signatures of both host and Symbiodinium changed with depth, indicating that coral colonies acquired energy from different sources depending on depth. This study highlights the complexity in physiological adaptations within this symbiosis and the different strategies used by closely related mesophotic species to diversify energy acquisition and to successfully establish and compete in extreme light‐limited environments.

Transcriptomic analyses highlight the likely metabolic consequences of colonization of a cnidarian host by native or non-native Symbiodinium species.

ABSTRACTReef-building corals and some other cnidarians form symbiotic relationships with members of the dinoflagellate family Symbiodinaceae. As Symbiodinaceae is a highly diverse taxon, the physiological interactions between its members and their hosts are assumed to differ between associations. The presence of different symbiont types is known to affect expression levels of specific host genes, but knowledge of the effects on the transcriptome more broadly remains limited. In the present study, transcriptome profiling was conducted on the tropical corallimorpharian, Ricordea yuma, following the establishment of symbiosis with either the ‘homologous’ symbiont Symbiodinium goreaui (also known as Cladocopium goreaui; ITS2 type C1) or ‘heterologous’ symbionts (predominantly S. trenchii, which is also known as Durusdinium trenchii; ITS2 type D1a) isolated from a different corallimorpharian host (Rhodactis indosinensis). Transcriptomic analyses showed that genes encoding host glycogen biosynthesis pathway components are more highly induced during colonization by the homologous symbiont than by the heterologous symbiont. Similar patterns were also observed for several other genes thought to facilitate symbiotic nutrient exchange, including those involved in lipid translocation/storage and metabolite transport. The gene expression results presented here imply that colonization by homologous or heterologous Symbiodinium types may have very different metabolic consequences for the Ricordea host, supporting the notion that even though some cnidarians may be able to form novel symbioses after bleaching, the metabolic performance of these may be compromised.This article has an associated First Person interview with the first author of the paper.

Symbiont type and environmental factors affect transcriptome-wide gene expression in the coral Montipora capitata.

Reef‐building corals may harbor genetically distinct lineages of endosymbiotic dinoflagellates in the genus Symbiodinium, which have been shown to affect important colony properties, including growth rates and resilience against environmental stress. However, the molecular processes underlying these differences are not well understood. In this study, we used whole transcriptome sequencing (RNA‐seq) to assess gene expression differences between 27 samples of the coral Montipora capitata predominantly hosting two different Symbiodinium types in clades C and D. The samples were further characterized by their origin from two field sites on Hawai‘i Island with contrasting environmental conditions. We found that transcriptome‐wide gene expression profiles clearly separated by field site first, and symbiont clade second. With 273 differentially expressed genes (DEGs, 1.3% of all host transcripts), symbiont clade had a measurable effect on host gene expression, but the effect of field site proved almost an order of magnitude higher (1,957 DEGs, 9.6%). According to SNP analysis, we found moderate evidence for host genetic differentiation between field sites (F ST = 0.046) and among corals harboring alternative symbiont clades (F ST = 0.036), suggesting that site‐related gene expression differences are likely due to a combination of local adaptation and acclimatization to environmental factors. The correlation between host gene expression and symbiont clade may be due to several factors, including host genotype or microhabitat selecting for alternative clades, host physiology responding to different symbionts, or direct modulation of host gene expression by Symbiodinium. However, the magnitude of these effects at the level of transcription was unexpectedly small considering the contribution of symbiont type to holobiont phenotype.

Contrasting effects of Symbiodinium identity on coral host transcriptional profiles across latitudes.

Reef-building corals can increase their resistance to heat-induced bleaching through adaptation and acclimatization and/or by associating with a more thermo-tolerant strain of algal symbiont (Symbiodinium sp.). Here, we show that these two adaptive pathways interact. We collected Acropora millepora corals from two contrasting thermal environments on the Great Barrier Reef: cooler, mid-latitude Orpheus Island, where all corals hosted a heat-sensitive clade C Symbiodinium, and warmer, low-latitude Wilkie Island, where corals hosted either a clade C or a more thermo-tolerant clade D. Corals were kept in a benign common garden to reveal differences in baseline gene expression, reflecting prior adaptation/long-term acclimatization. Model-based analysis identified gene expression differences between Wilkie and Orpheus corals that were negatively correlated with previously described transcriptome-wide signatures of heat stress, signifying generally elevated thermotolerance of Wilkie corals. Yet, model-free analyses of gene expression revealed that Wilkie corals hosting clade C were distinct from Wilkie corals hosting clade D, whereas Orpheus corals were more variable. Wilkie corals hosting clade C symbionts exhibited unique functional signatures, including downregulation of histone proteins and ion channels and upregulation of chaperones and RNA processing genes, putatively representing constitutive "frontloading" of stress response genes. Furthermore, clade C Symbiodinium exhibited constitutive expression differences between Wilkie and Orpheus, indicative of contrasting life history strategies. Our results demonstrate that hosting alternative Symbiodinium types is associated with different pathways of local adaptation for the coral host. These interactions could play a significant role in setting the direction of genetic adaptation to global warming in the two symbiotic partners.

Experimental Evolution in Coral Photosymbionts as a Tool to Increase Thermal Tolerance

Coral reefs are under major threat from ocean warming. When temperatures become too high corals bleach, expelling their symbiotic, photosynthetic microalgae (Symbiodinium), which they depend on for much of their nutritional requirements. Prolonged bleaching has led to widespread coral mortality and the severity and frequency of bleaching events are predicted to increase in the future. Coral bleaching tolerance is influenced by the thermal tolerance of the Symbiodinium harboured, and these microbial members of the coral holobiont may be able to evolve more rapidly than the coral host itself. Here, we examined the response of replicate cultures of five genetically distinct Symbiodinium strains (A3c, two types of D1, G3, and F1) to increasing temperatures over the course of approximately one year. For three Symbiodinium types (types A3c, G3, and F1), we observed a stable adaptive change at the end of this exposure period, which equated to only 41-69 asexual generations. The long-term selected Symbiodinium culture replicates (SS) showed faster growth rates under short-term, acute heat stress, and in some cases higher photosynthetic efficiencies, compared to wild-type populations (WT). Our results considerably extend the field of experimental evolution in Symbiodinium and with further work into the Symbiodinium-coral association and bleaching response, this approach may become a valuable tool in coral reef conservation and restoration initiatives.

Molecular signatures of host specificity linked to habitat specialization in Exaiptasia sea anemones.

Rising ocean temperatures associated with global climate change induce breakdown of the symbiosis between coelenterates and photosynthetic microalgae of the genus Symbiodinium. Association with more thermotolerant partners could contribute to resilience, but the genetic mechanisms controlling specificity of hosts for particular Symbiodinium types are poorly known. Here, we characterize wild populations of a sea anemone laboratory model system for anthozoan symbiosis, from contrasting environments in Caribbean Panama. Patterns of anemone abundance and symbiont diversity were consistent with specialization of holobionts for particular habitats, with Exaiptasia pallida/S. minutum (ITS2 type B1) abundant on vertical substrate in thermally stable, shaded environments but E. brasiliensis/Symbiodinium sp. (ITS2 clade A) more common in shallow areas subject to high temperature and irradiance. Population genomic sequencing revealed a novel E. pallida population from the Bocas del Toro Archipelago that only harbors S. minutum. Loci most strongly associated with divergence of the Bocas‐specific population were enriched in genes with putative roles in cnidarian symbiosis, including activators of the complement pathway of the innate immune system, thrombospondin‐type‐1 repeat domain proteins, and coordinators of endocytic recycling. Our findings underscore the importance of unmasking cryptic diversity in natural populations and the role of long‐term evolutionary history in mediating interactions with Symbiodinium.

Dynamics of coral-associated microbiomes during a thermal bleaching event.

Coral‐associated microorganisms play an important role in their host fitness and survival. A number of studies have demonstrated connections between thermal tolerance in corals and the type/relative abundance of Symbiodinium they harbor. More recently, the shifts in coral‐associated bacterial profiles were also shown to be linked to the patterns of coral heat tolerance. Here, we investigated the dynamics of Porites lutea‐associated bacterial and algal communities throughout a natural bleaching event, using full‐length 16S rRNA and internal transcribed spacer sequences (ITS) obtained from PacBio circular consensus sequencing. We provided evidence of significant changes in the structure and diversity of coral‐associated microbiomes during thermal stress. The balance of the symbiosis shifted from a predominant association between corals and Gammaproteobacteria to a predominance of Alphaproteobacteria and to a lesser extent Betaproteobacteria following the bleaching event. On the contrary, the composition and diversity of Symbiodinium communities remained unaltered throughout the bleaching event. It appears that the switching and/or shuffling of Symbiodinium types may not be the primary mechanism used by P. lutea to cope with increasing seawater temperature. The shifts in the structure and diversity of associated bacterial communities may contribute more to the survival of the coral holobiont under heat stress.

Using Seawater to Document Coral-Zoothanthella Diversity: A New Approach to Coral Reef Monitoring Using Environmental DNA

Frequent, high-density coral monitoring is essential to understand coral reef ecosystems. For this purpose, we developed a novel method for simultaneous monitoring of Acropora corals and their symbiont, Symbiodinium, from environmental DNA (eDNA) in seawater using next generation sequencing technology (NGS). We performed a tank experiment with running seawater using 19 Acropora species. Complete mitochondrial genomes of all the Acropora species were assembled to create a database and major types of their Symbiodinium symbionts were identified. Then eDNA was isolated by filtering inlet and outlet seawater from the tanks. Acropora and Symbiodinium DNA were amplified by PCR and sequenced. We detected all of the tested Acropora types from eDNA samples. Proportions of the DNA sequences and the weights of corals, and the number of DNA sequences and the masses of corals were positively correlated in the tanks, respectively. In this trial, we detected DNA sequences from as little as 0.04 kg of Acropora colony, suggesting that existence of at least one adult Acropora colony (approx. 30 cm diameter = 1kg) per m2 at depths <10m could be detected using eDNA in the field. In addition, we detected major types of Symbiodinium within host corals from seawater, suggesting that it should be possible to detect major coral symbiont types if Acropora corals exist nearby, and possible free-living state Symbiodinium cells from eDNA in sea water. eDNA abundance of Symbiodinium types did not correlate well with frequencies of major Symbiodinium types in the corals, suggesting that quantification of Symbiodinium is difficult at this stage. Although this is the initial attempt to detect coral and Symbiodinium simultaneously from eDNA in seawater, this method may allow us to perform high-frequency, high-density coral reef monitoring of both corals and their symbionts in the near future.

Unexpected mixed-mode transmission and moderate genetic regulation of Symbiodinium communities in a brooding coral

Determining the extent to which Symbiodinium communities in corals are inherited versus environmentally acquired is fundamental to understanding coral resilience and to predicting coral responses to stressors like warming oceans that disrupt this critical endosymbiosis. We examined the fidelity with which Symbiodinium communities in the brooding coral Seriatopora hystrix are vertically transmitted and the extent to which communities are genetically regulated, by genotyping the symbiont communities within 60 larvae and their parents (9 maternal and 45 paternal colonies) using high-throughput sequencing of the ITS2 locus. Unexpectedly, Symbiodinium communities associated with brooded larvae were distinct from those within parent colonies, including the presence of types not detected in adults. Bayesian heritability (h2) analysis revealed that 33% of variability in larval Symbiodinium communities was genetically controlled. Results highlight flexibility in the establishment of larval symbiont communities and demonstrate that symbiont transmission is not exclusively vertical in brooding corals. Instead, we show that Symbiodinium transmission in S. hystrix involves a mixed-mode strategy, similar to many terrestrial invertebrate symbioses. Also, variation in the abundances of common Symbiodinium types among adult corals suggests that microhabitat differences influence the structure of in hospite Symbiodinium communities. Partial genetic regulation coupled with flexibility in the environmentally acquired component of Symbiodinium communities implies that corals with vertical transmission, like S. hystrix, may be more resilient to environmental change than previously thought.

Symbiont Identity Influences Patterns of Symbiosis Establishment, Host Growth, and Asexual Reproduction in a Model Cnidarian-Dinoflagellate Symbiosis.

The genus Symbiodinium is physiologically diverse and so may differentially influence symbiosis establishment and function. To explore this, we inoculated aposymbiotic individuals of the sea anemone Exaiptasia pallida (commonly referred to as "Aiptasia"), a model for coral symbiosis, with one of five Symbiodinium species or types (S. microadriaticum, S. minutum, phylotype C3, S. trenchii, or S. voratum). The spatial pattern of colonization was monitored over time via confocal microscopy, and various physiological parameters were measured to assess symbiosis functionality. Anemones rapidly formed a symbiosis with the homologous symbiont, S. minutum, but struggled or failed to form a long-lasting symbiosis with Symbiodinium C3 or S. voratum, respectively. Symbiodinium microadriaticum and S. trenchii were successful but reached their peak density two weeks after S. minutum. The spatial pattern of colonization was identical for all Symbiodinium taxa that were ultimately successful, starting in the oral disk and progressing to the tentacles, before invading the column and, finally, the pedal disk. In all cases, proliferation through the anemone's tentacles was patchy, suggesting that symbionts were being expelled into the gastrovascular cavity and re-phagocytosed by the host. However, the timing of these various spatial events differed between the different Symbiodinium taxa. Furthermore, S. microadriaticum and S. trenchii were less beneficial to the host, as indicated by lower rates of photosynthesis, anemone growth, and pedal laceration. This study enhances our understanding of the link between symbiont identity and the performance of the overall symbiosis, which is important for understanding the potential establishment and persistence of novel host-symbiont pairings. Importantly, we also provide a baseline for further studies on this topic with the globally adopted "Aiptasia" model system.

Symbiosis process between Acropora larvae and Symbiodinium differs even among closely related Symbiodinium types

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 592:119-128 (2018) - DOI: https://doi.org/10.3354/meps12474 Symbiosis process between Acropora larvae and Symbiodinium differs even among closely related Symbiodinium types Hiroshi Yamashita1,*, Go Suzuki1, Chuya Shinzato2,3, Mitsuru Jimbo4, Kazuhiko Koike5 1Research Center for Subtropical Fisheries, Seikai National Fisheries Research Institute, Japan Fisheries Research and Education Agency, Ishigaki, Okinawa 907-0451, Japan 2Marine Genomics Unit, Okinawa Institute of Science and Technology Graduate University, Onna, Okinawa 904-0495, Japan 3Atmosphere and Ocean Research Institute, The University of Tokyo, Kashiwa, Chiba 277-8564, Japan 4School of Marine Biosciences, Kitasato University, Sagamihara, Kanagawa 252-0373, Japan 5Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8528, Japan *Corresponding author: hyamashita@fra.affrc.go.jp ABSTRACT: The symbiosis between corals and the dinoflagellate genus Symbiodinium is a fundamental pillar of coral reef ecosystems. Many of the coral species, including Acropora, acquire Symbiodinium from the environment during early ontogeny for symbiosis. Genetically diverse members of Symbiodinium have been identified from the environment, but naturally settled Acropora recruits (i.e. settlers or spats) usually harbor specific members of Symbiodinium. Thus, symbiosis between Acropora and Symbiodinium is not established haphazardly in the wild. Conversely, under laboratory conditions, Acropora larvae can acquire other Symbiodinium varieties. However, it remains unclear whether a stable symbiosis between these Symbiodinium that are never detected in natural Acropora recruits and corals is successfully established. Here, we artificially supplied A. tenuis larvae with 3 closely related Symbiodinium culture strains, namely, type A1 (common within natural Acropora recruits), type A2-relative (never detected within corals), and type A3 (often found within natural Acropora recruits). We then determined the percentage of Symbiodinium-infected larvae and the density of Symbiodinium cells infecting the larvae. For types A1 and A3, the percentage of Symbiodinium-infected larvae and the density of Symbiodinium cells did not decrease even after stopping the Symbiodinium supply. This result indicated that stable symbiosis was successfully established, even in the laboratory. However, the percentage of Symbiodinium-infected larvae and Symbiodinium cell density of type A2-relative declined after stopping the Symbiodinium supply. Thus, under laboratory conditions, A. tenuis larvae can acquire Symbiodinium that is never detected within natural corals; however, stable symbiosis between these Symbiodinium and corals was not successfully established. KEY WORDS: Early symbiosis · Symbiodinium · Zooxanthellae · Acropora · Coral larvae Full text in pdf format PreviousNextCite this article as: Yamashita H, Suzuki G, Shinzato C, Jimbo M, Koike K (2018) Symbiosis process between Acropora larvae and Symbiodinium differs even among closely related Symbiodinium types. Mar Ecol Prog Ser 592:119-128. https://doi.org/10.3354/meps12474 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 592. Online publication date: March 29, 2018 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2018 Inter-Research.

Temperature and Water Quality-Related Patterns in Sediment-Associated Symbiodinium Communities Impact Symbiont Uptake and Fitness of Juveniles in the Genus Acropora

The majority of corals acquire their photo-endosymbiont Symbiodinium from environmental sources anew each generation. Despite the critical role that environmental availability of Symbiodinium plays in the potential for corals to acclimate and adapt to changing environments, little is known about the diversity of free-living Symbiodinium communities and how variation in these communities influences uptake and in hospite communities in juvenile corals. Here we characterize Symbiodinium community diversity in sediment samples collected from eight reefs representing latitudinal and cross-shelf variation in water quality and temperature regimes. Sediment-associated Symbiodinium communities were then compared to in hospite communities acquired by A. tenuis and A. millepora juveniles following 11 – 145 days of experimental exposure to sediments from each of the reefs. Communities associated with juveniles and sediments differed substantially, with sediments harbouring four times more unique OTUs than juveniles (1125 OTUs vs. 271). Moreover, only 10.6% of these OTUs were shared between juveniles and sediments, indicating selective uptake by acroporid juveniles. The diversity and abundance of Symbiodinium types differed among sediment samples from different temperature and water quality environments. Symbiodinium communities acquired by juveniles also differed among the sediment treatments, despite juveniles having similar parentage. Moreover, Symbiodinium communities displayed different rates of infection, mortality, and photochemical efficiencies, important traits for coral fitness. This study demonstrates that the biogeography of free-living Symbiodinium types found within sediment reservoirs follows patterns along latitudinal and water quality environmental gradients on the Great Barrier Reef. We also demonstrate a bipartite strategy for Symbiodinium uptake by juvenile corals of two horizontally-transmitting acroporid species, whereby uptake is selective within the constraints of environmental availability.

Shift of symbiont communities in Acropora tenuis juveniles under heat stress.

Ocean warming is a major threat to coral reefs, leading to an increasing frequency and amplitude of coral bleaching events, where the coral and its algal symbiont associations breakdown. Long-term change and resilience of a symbiont community in coral juveniles is thought to be one of the most important aspects for determining thermal tolerance of the coral holobionts; however, despite its importance, they are not well documented in both under elevated temperature and even under natural condition. Here we investigated changes in symbiont communities in juveniles of the coral Acropora tenuis under controlled heat stress conditions (30 °C, 31/32 °C) and natural variations in seawater temperatures (19–30 °C) for up to four months. Compared with the ambient temperature conditions, coral survival rates were higher when exposed to 30 °C, but survival rates decreased when exposed to 31/32 °C. Symbiodinium types A3, C1, and D1-4 were detected in the juveniles under all thermal conditions; however, in higher water temperatures (31/32 °C), both the prevalence of D1-4 Symbiodinium and the number of juveniles harboring only this type of symbiont increased after two to four months later. In contrast, colonies at lower temperatures (30 °C and ambient) harbored multiple clades of symbionts over the same experimental period. These results highlight the flexibility of the coral–Symbiodinium symbiosis for juvenile A. tenuis under variable thermal conditions. In particular, the benefit of the preferential association with type D1-4 can be considered as a response when under heat-stress conditions, and that could help corals to cope with ocean warming.

Bleaching in reef invertebrate larvae associated with Symbiodinium strains within clades A–F

Coral reefs are diverse ecosystems relying on the presence of dinoflagellates (genus Symbiodinium), that are found in symbiotic association with multiple phyla and performing the majority of primary production. However, coral reefs are currently threatened by climate change and the increase in seawater temperature, which causes the bleaching phenomenon. While bleaching has been well documented for adult host organisms, it is still poorly understood in larval stages. We offered Symbiodinium types within clades A–F to the larvae of Mussismilia hispida (scleractinian coral), Berghia stephanieae (nudibranch) and Tridacna crocea (giant clam) and manipulated the temperature to 26, 29 or 32 °C. Samples were taken at 0, 12, 24, 48 and 72 h post-temperature increase, chlorophyll-a (chl-a) was extracted and its content measured in a fluorometer. Symbiodinium type, temperature and time all influenced chl-a content. M. hispida larvae displayed a bleaching threshold at 29 °C; larvae containing Symbiodinium A–F all bleached at 32 °C, but with significantly lower bleaching in larvae associated with type A1. B. stephanieae digested the symbionts; while chl-a content decreased over time equally for all clades, it is not possible to determine if it is related to bleaching. T. crocea larvae at 29 °C bleached for all symbiont types, except for A1. At 32 °C, all types were bleached, but type A1 bleached significantly less. These findings show that type A1 seems to be more thermo-tolerant in larvae of the tested species. This may be related to the fact that strains within this clade are homologous to both M. hispida and T. crocea, as they are found within these adult host’ tissues. Therefore, symbiont type may have an important role in invertebrate larvae development and present relevant implications for recruitment.

Production of three symbiosis-related fatty acids by Symbiodinium types in clades A–F associated with marine invertebrate larvae

Symbiodinium are dinoflagellates engaged in a mutualistic symbiosis with multiple coral reef taxa. They are divided in nine different clades (A–I), which typically associate with different hosts. However, very little information is available on metabolic differences in Symbiodinium types, especially when associated with metazoan larvae. We tested whether three ω3 fatty acids (stearidonic acid, SDA; docosapentaenoic acid, DPA; and docosahexaenoic acid, DHA) that are typically translocated from Symbiodinium to its host are produced by Symbiodinium types within clades A–F associated with Mussismilia hispida (scleractinian coral), Berghia stephanieae (nudibranch), and Tridacna crocea (giant clam) larvae. We acquired and spawned broodstock for each host, cultured their larvae, and offered Symbiodinium types belonging to clades A–F. Samples were taken during a 72-h window after the offer of Symbiodinium, and fatty acids were extracted and analyzed by gas chromatography. The concentrations of SDA and DPA for all host larvae–dinoflagellate associations were low and variable, without trends. However, M. hispida planula larvae associated with Symbiodinium A1 and C1 had a statistically significant higher amount of DHA. The veliger larvae of B. stephanieae digested the Symbiodinium, and the amount of DHA remained constant throughout the experiment. The veliger larvae of T. crocea associated with Symbiodinium A1 and C1 also presented a higher amount of DHA, although not statistically different from the other types. These results show that Symbiodinium A1 and C1, in the case of M. hispida and T. crocea (which usually harbor strains within clades A and C), may contribute a small amount of DHA to the larvae of these organisms and form a stronger mutualism than other strains.

Natural Variation in Responses to Acute Heat and Cold Stress in a Sea Anemone Model System for Coral Bleaching.

Rising ocean temperatures disrupt the symbiosis between corals and their microalgae, accelerating global decline of coral reef ecosystems. Because of the difficulty of performing laboratory experiments with corals, the sea anemone Aiptasia has emerged as an important model system for molecular studies of coral bleaching and symbiosis. Here, we investigate natural variation in bleaching responses among different genetic lineages of Aiptasia. Both heat- and cold-induced paths to symbiosis breakdown were analyzed. Significant genetic variation in response to acute heat stress was observed, with severe bleaching of two Aiptasia strains from Hawaii but minimal bleaching of strains from the U.S. South Atlantic, including the strain used to generate the Aiptasia reference genome. Both strains from Hawaii hosted Symbiodinium type B1, whereas strains from the U.S. South Atlantic hosted type A4 or B2. In contrast to the results from exposures to acute heat stress, negligible variation was observed in response to a pulsed cold shock despite moderate bleaching across all strains. These results support our hypothesis that bleaching responses to distinct stressors are independent. Our findings emphasize the role of stress regime when predicting adaptive responses of symbiotic cnidarians to changing climates, because genetic variation may exist for some forms of stress-induced bleaching but not others.

Seriatopora Diversity Preserved in Upper Mesophotic Coral Ecosystems in Southern Japan

Coral reefs worldwide are facing increasing stress due to drastic changes in their environment. Mesophotic coral ecosystems (MCEs) have been considered as a potential refuge from several major stressors, such as warm-water bleaching events. However, their role as a subsequent source of larvae remains unclear for many species, particularly as genetic differentiation of corals over depth has frequently been observed. In 1998 and 2001, two severe bleaching events around Okinawa Island in Japan resulted in major changes to the shallow reefs, including the local “extinction” of species such as Seriatopora hystrix at Sesoko Island. However, recently this species was found to be present in abundance at mesophotic depths in the area, despite no clear signs of recovery being observed in the adjacent shallow waters. Here, we assessed the genetic diversity of Seriatopora from this deep population and provide a comparison with populations from shallow to mesophotic depths in other parts of the Ryukyu archipelago, to understand their depth specificity and their importance in genetic diversity conservation of affected shallow populations. High levels of genetic diversity were observed in both shallow and mesophotic Seriatopora populations for both the nuclear (internal transcribed spacer 2, ITS2) and the mitochondrial (hypervariable open reading frame, ORF) marker, with no clear partitioning of haplotypes over depth or across locations in the archipelago. Both ITS2 and ORF suggest the presence of potential cryptic species and the discrepancy between the markers could reflect hybridization or incomplete lineage sorting. Although associated endosymbionts (Symbiodinium spp.) all shared the same mitochondrial haplotype (cytochrome oxidase subunit 1, COI), the nuclear ribosomal ITS2 revealed slight potential habitat partitioning between the genotypes, with a small decrease of C59 Symbiodinium types below 10 m depth and a mirrored increase in C1/C78a-related types. The relative absence of depth-specific host lineages and the substantial overlap between shallow and deep Symbiodinium types indicate that Okinawan MCEs may act as a refuge preserving genotypic diversity of bleaching-sensitive coral such as Seriatopora, and on the long-term may have the potential to contribute to shallow-water recolonization of this species.

Expression of a symbiosis-specific gene in Symbiodinium type A1 associated with coral, nudibranch and giant clam larvae.

Symbiodinium are responsible for the majority of primary production in coral reefs and found in a mutualistic symbiosis with multiple animal phyla. However, little is known about the molecular signals involved in the establishment of this symbiosis and whether it initiates during host larval development. To address this question, we monitored the expression of a putative symbiosis-specific gene (H+-ATPase) in Symbiodinium A1 ex hospite and in association with larvae of a scleractinian coral (Mussismilia hispida), a nudibranch (Berghia stephanieae) and a giant clam (Tridacna crocea). We acquired broodstock for each host, induced spawning and cultured the larvae. Symbiodinium cells were offered and larval samples taken for each host during the first 72 h after symbiont addition. In addition, control samples including free-living Symbiodinium and broodstock tissue containing symbionts for each host were collected. RNA extraction and RT-PCR were performed and amplified products cloned and sequenced. Our results show that H+-ATPase was expressed in Symbiodinium associated with coral and giant clam larvae, but not with nudibranch larvae, which digested the symbionts. Broodstock tissue for coral and giant clam also expressed H+-ATPase, but not the nudibranch tissue sample. Our results of the expression of H+-ATPase as a marker gene suggest that symbiosis between Symbiodinium and M. hispida and T. crocea is established during host larval development. Conversely, in the case of B. stephanieae larvae, evidence does not support a mutualistic relationship. Our study supports the utilization of H+-ATPase expression as a marker for assessing Symbiodinium–invertebrate relationships with applications for the differentiation of symbiotic and non-symbiotic associations. At the same time, insights from a single marker gene approach are limited and future studies should direct the identification of additional symbiosis-specific genes, ideally from both symbiont and host.

Carbon and Nitrogen Acquisition in Shallow and Deep Holobionts of the Scleractinian Coral S. pistillata

Reef building corals can host different symbiont genotypes (clades), and form distinct holobionts in response to environmental changes. Studies on the functional significance of genetically different symbionts have focused on the thermal tolerance rather than on the nutritional significance. Here, we characterized the nitrogen and carbon assimilation rates, the allocation patterns of these nutrients within the symbiosis, and the trophic condition of two distinct holobionts of Stylophora pistillata: one associated with Symbiodinium clade A in shallow reefs and the other one associated with clade C in mesophotic reefs. The main findings are that: (1) clade C-symbionts have a competitive advantage for the acquisition of carbon at low irradiance compared to clade A-symbionts; (2) light is however the primary factor that determines the positive relationship between the amount of carbon fixed in photosynthesis by the symbionts and the amount of carbon translocated to the host; and (3) by contrast, the dominant Symbiodinium type preferentially determines a negative relationship between rates of coral feeding and nitrogen assimilation, although light still plays a relevant role in this relationship. Clade C-holobionts had indeed higher heterotrophic capacities, but lower inorganic nitrogen assimilation rates than clade A-holobionts, at all light levels. Broadly, our results show that the assimilation and translocation rates of inorganic carbon and nitrogen are clade and light-dependent. In addition, the capacity of S. pistillata to form mesophotic reefs in the Red Sea relies on its ability to select Symbiodinium clade C, as this symbiont type is more efficient to fix carbon at low light.