Defending against pathogens - immunological priming and its molecular basis in a sea anemone, cnidarian

Cnidarians, in general, are long-lived organisms and hence may repeatedly encounter common pathogens during their lifespans. It remains unknown whether these early diverging animals possess some type of immunological reaction that strengthens the defense response upon repeated infections, such as that described in more evolutionary derived organisms. Here we show results that sea anemones that had previously encountered a pathogen under sub-lethal conditions had a higher survivorship during a subsequently lethal challenge than naive anemones that encountered the pathogen for the first time. Anemones subjected to the lethal challenge two and four weeks after the sub-lethal exposure presented seven- and five-fold increases in survival, respectively, compared to the naive anemones. However, anemones challenged six weeks after the sub-lethal exposure showed no increase in survivorship. We argue that this short-lasting priming of the defense response could be ecologically relevant if pathogen encounters are restricted to short seasons characterized by high stress. Furthermore, we discovered significant changes in proteomic profiles between naive sea anemones and those primed after pathogen exposure suggesting a clear molecular signature associated with immunological priming in cnidarians. Our findings reveal that immunological priming may have evolved much earlier in the tree of life than previously thought

Recurring Episodes of Thermal Stress Shift the Balance from a Dominant Host-Specialist to a Background Host-Generalist Zooxanthella in the Threatened Pillar Coral, Dendrogyra cylindrus

Most scleractinian corals form obligate symbioses with photosynthetic dinoflagellates (family Symbiodiniaceae), which provide differential tolerances to their host. Previously, research has focused on the influence of symbiont composition and the dynamic processes of symbiont repopulation during single episodes of hyperthermal events, followed by years of less-stressful conditions. In contrast, this study characterized for the first time, the role of Symbiodiniaceae species changes in response to annually recurring hyperthermal events, a scenario soon expected to become the norm. Consecutive hyperthermal events during summer 2014 and 2015 along the Florida Reef Tract offered a unique opportunity to study bleaching susceptibility and recovery under recurrent annual hyperthermal scenarios. We utilized Illumina amplicon sequencing of the chloroplast 23S DNA region to assess with fine resolution the Symbiodiniaceae diversity associated with pillar coral, Dendrogyra cylindrus. Our findings show diverse assemblages of Symbiodiniaceae species and that some cryptic members are not transient associates but persistent and ecologically relevant, especially during recurrent annual warming events. This was evidenced by changes in relative abundance from the typically dominant host-specialist endosymbiont, Breviolum dendrogyrum, to B. meandrinium a host-generalist species common to corals in the family Meandrinidae but occurs at background densities in most coral colonies of D. cylindrus. The rise in abundance of B. meandrinium associated strongly with bleaching resistance in the coral host during two consecutive hyperthermal events. In some cases, host-compatible background symbionts can rapidly increase in abundance during episodes of stress and may impart physiological resilience to rapid environmental change; and thus, represents a potentially important ecological process by which symbiotic corals acclimatize to changing ocean conditions

Transcriptome analysis of a cnidarian – dinoflagellate mutualism reveals complex modulation of host gene expression

BACKGROUND: Cnidarian – dinoflagellate intracellular symbioses are one of the most important mutualisms in the marine environment. They form the trophic and structural foundation of coral reef ecosystems, and have played a key role in the evolutionary radiation and biodiversity of cnidarian species. Despite the prevalence of these symbioses, we still know very little about the molecular modulators that initiate, regulate, and maintain the interaction between these two different biological entities. In this study, we conducted a comparative host anemone transcriptome analysis using a cDNA microarray platform to identify genes involved in cnidarian – algal symbiosis. RESULTS: We detected statistically significant differences in host gene expression profiles between sea anemones (Anthopleura elegantissima) in a symbiotic and non-symbiotic state. The group of genes, whose expression is altered, is diverse, suggesting that the molecular regulation of the symbiosis is governed by changes in multiple cellular processes. In the context of cnidarian – dinoflagellate symbioses, we discuss pivotal host gene expression changes involved in lipid metabolism, cell adhesion, cell proliferation, apoptosis, and oxidative stress. CONCLUSION: Our data do not support the existence of symbiosis-specific genes involved in controlling and regulating the symbiosis. Instead, it appears that the symbiosis is maintained by altering expression of existing genes involved in vital cellular processes. Specifically, the finding of key genes involved in cell cycle progression and apoptosis have led us to hypothesize that a suppression of apoptosis, together with a deregulation of the host cell cycle, create a platform that might be necessary for symbiont and/or symbiont-containing host cell survival. This first comprehensive molecular examination of the cnidarian – dinoflagellate associations provides critical insights into the maintenance and regulation of the symbiosis

Proteomic Basis of Symbiosis: A Heterologous Partner Fails to Duplicate Homologous Colonization in a Novel Cnidarian– Symbiodiniaceae Mutualism

Reef corals and sea anemones form symbioses with unicellular symbiotic dinoflagellates. The molecular circumventions that underlie the successful intracellular colonization of hosts by symbionts are still largely unknown. We conducted proteomic analyses to determine molecular differences of Exaiptasia pallida anemones colonized by physiologically different symbiont species, in comparison with symbiont-free (aposymbiotic) anemones. We compared one homologous species, Symbiodinium linucheae, that is natively associated with the clonal Exaiptasia strain (CC7) to another heterologous species, Durusdinium trenchii, a thermally tolerant species that colonizes numerous coral species. This approach allowed the discovery of a core set of host genes that are differentially regulated as a function of symbiosis regardless of symbiont species. The findings revealed that symbiont colonization at higher densities requires circumvention of the host cellular immunological response, enhancement of ammonium regulation, and suppression of phagocytosis after a host cell in colonized. Furthermore, the heterologous symbionts failed to duplicate the same level of homologous colonization within the host, evidenced by substantially lower symbiont densities. This reduced colonization of D. trenchii correlated with its inability to circumvent key host systems including autophagy-suppressing modulators, cytoskeletal alteration, and isomerase activity. The larger capability of host molecular circumvention by homologous symbionts could be the result of a longer evolutionary history of host/symbiont interactions, which translates into a more finely tuned symbiosis. These findings are of great importance within the context of the response of reef corals to climate change since it has been suggested that coral may acclimatize to ocean warming by changing their dominant symbiont species

Recurring Episodes of Thermal Stress Shift the Balance From a Dominant Host-Specialist to a Background Host-Generalist Zooxanthella in the Threatened Pillar Coral, Dendrogyra cylindrus

Most scleractinian corals form obligate symbioses with photosynthetic dinoflagellates (family Symbiodiniaceae), which provide differential tolerances to their host. Previously, research has focused on the influence of symbiont composition and the dynamic processes of symbiont repopulation during single episodes of hyperthermal events, followed by years of less-stressful conditions. In contrast, this study characterized for the first time, the role of Symbiodiniaceae species changes in response to annually recurring hyperthermal events, a scenario soon expected to become the norm. Consecutive hyperthermal events during summer 2014 and 2015 along the Florida Reef Tract offered a unique opportunity to study bleaching susceptibility and recovery under recurrent annual hyperthermal scenarios. We utilized Illumina amplicon sequencing of the chloroplast 23S DNA region to assess with fine resolution the Symbiodiniaceae diversity associated with pillar coral, Dendrogyra cylindrus. Our findings show diverse assemblages of Symbiodiniaceae species and that some cryptic members are not transient associates but persistent and ecologically relevant, especially during recurrent annual warming events. This was evidenced by changes in relative abundance from the typically dominant host-specialist endosymbiont, Breviolum dendrogyrum, to B. meandrinium a host-generalist species common to corals in the family Meandrinidae but occurs at background densities in most coral colonies of D. cylindrus. The rise in abundance of B. meandrinium associated strongly with bleaching resistance in the coral host during two consecutive hyperthermal events. In some cases, host-compatible background symbionts can rapidly increase in abundance during episodes of stress and may impart physiological resilience to rapid environmental change and thus, represents a potentially important ecological process by which symbiotic corals acclimatize to changing ocean conditions

Transcriptional Activation of c3 and hsp70 as Part of the Immune Response of Acropora millepora to Bacterial Challenges

The impact of disease outbreaks on coral physiology represents an increasing concern for the fitness and resilience of reef ecosystems. Predicting the tolerance of corals to disease relies on an understanding of the coral immune response to pathogenic interactions. This study explored the transcriptional response of two putative immune genes (c3 and c-type lectin) and one stress response gene (hsp70) in the reef building coral, Acropora millepora challenged for 48 hours with bacterial strains, Vibrio coralliilyticus and Alteromonas sp. at concentrations of 106 cells ml-1. Coral fragments challenged with V. coralliilyticus appeared healthy while fragments challenged with Alteromonas sp. showed signs of tissue lesions after 48 hr. Coral-associated bacterial community profiles assessed using denaturing gradient gel electrophoresis changed after challenge by both bacterial strains with the Alteromonas sp. treatment demonstrating the greatest community shift. Transcriptional profiles of c3 and hsp70 increased at 24 hours and correlated with disease signs in the Alteromonas sp. treatment. The expression of hsp70 also showed a significant increase in V. coralliilyticus inoculated corals at 24 h suggesting that even in the absence of disease signs, the microbial inoculum activated a stress response in the coral. C-type lectin did not show a response to any of the bacterial treatments. Increase in gene expression of c3 and hsp70 in corals showing signs of disease indicates their potential involvement in immune and stress response to microbial challenges

Building consensus around the assessment and interpretation of Symbiodiniaceae diversity

Within microeukaryotes, genetic variation and functional variation sometimes accumulate more quickly than morphological differences. To understand the evolutionary history and ecology of such lineages, it is key to examine diversity at multiple levels of organization. In the dinoflagellate family Symbiodiniaceae, which can form endosymbioses with cnidarians (e.g., corals, octocorals, sea anemones, jellyfish), other marine invertebrates (e.g., sponges, molluscs, flatworms), and protists (e.g., foraminifera), molecular data have been used extensively over the past three decades to describe phenotypes and to make evolutionary and ecological inferences. Despite advances in Symbiodiniaceae genomics, a lack of consensus among researchers with respect to interpreting genetic data has slowed progress in the field and acted as a barrier to reconciling observations. Here, we identify key challenges regarding the assessment and interpretation of Symbiodiniaceae genetic diversity across three levels: species, populations, and communities. We summarize areas of agreement and highlight techniques and approaches that are broadly accepted. In areas where debate remains, we identify unresolved issues and discuss technologies and approaches that can help to fill knowledge gaps related to genetic and phenotypic diversity. We also discuss ways to stimulate progress, in particular by fostering a more inclusive and collaborative research community. We hope that this perspective will inspire and accelerate coral reef science by serving as a resource to those designing experiments, publishing research, and applying for funding related to Symbiodiniaceae and their symbiotic partnerships.journal articl

Major Cellular and Physiological Impacts of Ocean Acidification on a Reef Building Coral

As atmospheric levels of CO2 increase, reef-building corals are under greater stress from both increased sea surface temperatures and declining sea water pH. To date, most studies have focused on either coral bleaching due to warming oceans or declining calcification due to decreasing oceanic carbonate ion concentrations. Here, through the use of physiology measurements and cDNA microarrays, we show that changes in pH and ocean chemistry consistent with two scenarios put forward by the Intergovernmental Panel on Climate Change (IPCC) drive major changes in gene expression, respiration, photosynthesis and symbiosis of the coral, Acropora millepora, before affects on biomineralisation are apparent at the phenotype level. Under high CO2 conditions corals at the phenotype level lost over half their Symbiodinium populations, and had a decrease in both photosynthesis and respiration. Changes in gene expression were consistent with metabolic suppression, an increase in oxidative stress, apoptosis and symbiont loss. Other expression patterns demonstrate upregulation of membrane transporters, as well as the regulation of genes involved in membrane cytoskeletal interactions and cytoskeletal remodeling. These widespread changes in gene expression emphasize the need to expand future studies of ocean acidification to include a wider spectrum of cellular processes, many of which may occur before impacts on calcification

Coral nursery water temperature during bleaching events in the Florida Reef Tract from 2014-2015 (EMUCoReS project)

Dataset: Acropora cervicornis nursery water temperaturesCoral nursery ("the bowl") water temperatures during summer bleaching events in 2014 and 2015. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/640227NSF Division of Ocean Sciences (NSF OCE) OCE-1503483, NSF Division of Ocean Sciences (NSF OCE) OCE-150343