Symbiotic Algae Research Articles

Microbiome Structuring Within a Coral Colony and Along a Sedimentation Gradient

Reef-building corals form complex relationships with a wide range of microbial partners, including symbiotic algae in the family Symbiodiniaceae and various bacteria. These coral-associated communities can be shaped to varying degrees by environmental context. Sedimentation can structure a coral’s microbial community by altering light availability for symbiotic algae, triggering the coral’s stress response, or serving as a reservoir for both pathogenic and essential bacterial and algal symbionts. To examine the influence of sedimentation on a coral’s microbiome, we used 16S rDNA and ITS-2 amplicon sequencing to characterize the bacterial and algal communities associated with the massive scleractinian coral Porites lobata across pairs of sites along a naturally occurring sedimentation gradient in Fouha Bay, southern Guam. Additionally, we investigate the influence of proximity to sediment on the coral colony scale, by sampling from the edge and center of colonies as well as the nearby sediment. The P. lobata colonies associated with several different genotypes of Cladocopium C15 algal symbionts and often harbored different genotypes within a single colony. However, the different Cladocopium genotypes showed no structuring according to colony position or location along the sedimentation gradient. Bacterial communities were largely consistent across the sedimentation gradient, however, some rarer taxa were differentially abundant across sites. Planococcaceae shows higher abundance closer to the river mouth in coral colonies in both the edge and center of colonies. Peredibacter also shows high abundance near the river mouth but only in sediment and the edges of the colony. We find sediment plays a larger role structuring bacterial communities at the colony scale compared to a coral’s position along the sedimentation gradient. Edge communities look more similar to the sediment compared to the center communities and are also enriched in similar pathways such as those involved in nitrogen fixation. We also find center samples to be dominated by Endozoicomonas compared to the edge, supporting a role for this taxon in structuring bacterial communities and limiting bacterial diversity in coral colonies. Together these results show the differential impact sedimentation can have between sections of the coral colony microhabitat.

Comparison of the bleaching susceptibility of coral species by using minimal samples of live corals.

In massive bleaching events (losing symbiotic algae from corals), more sensitive corals are bleached earlier than other corals. To perform a comparison of bleaching susceptibility within and across coral species, a simple quantitative method is required. Accordingly, we present a laboratory-based method for comparing the bleaching susceptibility of various coral species by using a standardized image analysis protocol. Coral fragments were sampled from the colonies of five species selected from Kenting, southern Taiwan, and maintained in the same aquarium tank with circulating seawater; 2 seawater temperature regimes were used (i.e., fast-heating program (FHP), with a heating rate of 1 °C per day; and slow-heating program (SHP), with a heating rate of 1 °C per 3 days). Each coral fragment was photographed periodically, and the colored images were subsequently converted to grayscale images and then digitally analyzed to determine the standardized grayscale values (G0) by comparing with that of standard color strip. The G0 of a sample at each time of photographing during bleaching was divided by the difference of G0 between the acclimating and the same but completely bleached fragment to derive the relative grayscale (RG%) at a particular stage of bleaching; this is done for each coral fragment of a colony. The smaller the RG% of a coral fragment the closer it is approaching completely bleached condition. The level of decrease in RG% within a time series of images in each heating regime was used to establish a bleaching time index (BTI). The lower the BTI, the sooner to reach a defined bleaching level (e.g., 30%), this indicates the coral is more sensitive to thermal bleaching. In the experiment, we compared the bleaching susceptibility of the five species. Based on the proposed BTI, the five species were ranked in terms of bleaching susceptibility, and the rankings were identical between the two temperature regimes; three species in Pocilloporidae had lower BTI, whereas the hydrocoral Millepora species had the highest BTI. Within each heating regime, the BTI of different species were ranked and used to indicate susceptibility. In the FHP, the three Pocilloporidae species could be divided into two groups in terms of bleaching susceptibility. FHP not only displayed a higher differentiating capability on coal bleaching susceptibility than SHP, but also had a faster completion time, thus reducing the likelihood of unforeseen complications during the tank experiments. Our color-based method is easier and less effort-intensive than methods involving the assessment of zooxanthellae densities. Moreover, it requires much fewer replicates and all samples in one large tank (e.g., 300 L) for the studies considering multiple species comparisons. This method opens opportunities for studying the effects of species types, acclimatization (e.g., seasons), and environmental factors other than temperature on coral bleaching.

Lipid profiling of coral symbiosomes in response to copper-induced carbon limitation: A metabolic effect of algal symbionts on the host immune status

Copper micropollutants are known to constrain coral's assimilation of carbonate, affecting the carbon available to algal symbionts and thus inducing a light stress. However, little is known regarding the physiological relevance of lipid metabolism in coral symbiotic algae in a carbon-limited state. Membrane lipids exhibit multiple physicochemical properties that are collectively responsible for the dynamic structure of cells depending on the physiological demands of the circumstances. To gain insight into lipid metabolism's importance in this regard, glycerophosphocholine (GPC) profiling of symbiosomes in coral (Seriatopora caliendrum) exposed to environmentally relevant copper levels (2.2–7.5 μg/L) for 4 days was performed in this study. Notably, reducing the number of 22:6-processing GPCs and increasing that of lyso-GPCs likely addressed the demands of metabolizing excess light energy, such as affecting the membrane dynamics to promote mitochondrial uncoupling. The decrease in 22:6-processing GPCs additionally protected cellular membranes from elevated oxidative stress, reducing their susceptibility to peroxidation and offsetting oxidized lipid-induced effects on membrane dynamics. The change in plasmanylcholines specifically localized within the symbiosome membrane also met the membrane requirements for responding to oxidative stress conditions. Moreover, increasing the 20:4-possessing plasmanylcholines and lysoplasmanylcholines and reducing the 22:6-possessing plasmanylcholines likely resulted in an imbalance of the immune reaction, influencing the coral–algae symbiosis given the role of such plasmanylcholines in cell signaling. In summary, carbon limitations induced by copper enrichment lead to a shift in the membrane lipid profile of coral symbiosomes, accommodating themselves to light stress conditions while compromising the symbiosis's stability.

Algal treatment of membrane rejects: a unique approach towards zero liquid discharge

A major concern in membrane-based water purification system is generation of huge concentrate stream and wastage of water. A typical Reverse osmosis (RO) or Nanofiltration (NF) system generates 20–25% reject containing high amount of dissolved salts and other contaminants. Contrary to popular belief, this reject water cannot be used without removing the contaminants or cannot be discharged anywhere. Main goal of this project is to find a cheapest and green way for treatment of RO/NF reject. Algal evaporation technique was explored in laboratory scale, to find its suitability for treatment of chloride-rich membrane reject in actual scenario and based on the results obtained, a pilot plant of 48KL was established on Hooghly Met Coke division (HMC), Tata Steel. Particular species of microalgae was selected, to take up minerals from reject water. There are several types of bacteria and symbiotic algae associated with selected micro algae survive in high TDS. A unique slope roof system, connected with algae growth tank, helps in efficient evaporation of water ensuring a Zero discharge. A markedly improved performance was achieved when algal evaporation followed solar evaporation. A total evaporation of 11 L/m2/day was observed, which was almost five times faster than Solar evaporation.

V-type H+-ATPase in the symbiosome membrane is a conserved mechanism for host control of photosynthesis in anthozoan photosymbioses.

In reef-building corals (order Scleractinia) and giant clams (phylum Molluca), V-type H+-ATPase (VHA) in host cells is part of a carbon concentrating mechanism (CCM) that regulates photosynthetic rates of their symbiotic algae. Here, we show that VHA plays a similar role in the sea anemone Anemonia majano, a member of the order Actinaria and sister group to the Scleractinia, which in contrast to their colonial calcifying coral relatives is a solitary, soft-bodied taxa. Western blotting and immunofluorescence revealed that VHA was abundantly present in the host-derived symbiosome membrane surrounding the photosymbionts. Pharmacological inhibition of VHA activity in individual anemones resulted in an approximately 80% decrease of photosynthetic O2 production. These results extend the presence of a host-controlled VHA-dependent CCM to non-calcifying cnidarians of the order Actiniaria, suggesting it is widespread among photosymbiosis between aquatic invertebrates and Symbiodiniaceae algae.

Black corals (Antipatharia) of the northern Red Sea: ancient predators of the mesophotic reef

Black corals (Antipatharia) are among the most common and diverse taxa in mesophotic and aphotic rocky habitats. Studies of their ecology and biology are limited mostly because of the technical challenges involved in deep-water work. Here, we describe taxonomic traits, habitat, and nutrition of 2 species from the upper mesophotic zone of the northern Red Sea, Gulf of Aqaba, Israel. Our study describes a broader distribution of the branched Antipathes griggi, which to date has been primarily found in the Hawaiian Archipelago. Additionally, a new record is reported regarding the coiled Stichopathes spp. discovered at the upper mesophotic zone. The branched A. griggi were more abundant than the coiled Stichopathes spp. in all explored sites, and differences in light and water flow conditions significantly affected their distribution. Both species lacked symbiotic algae (Symbiodiniaceae). Analysis of the amino acid compound-specific stable isotope signature (AA-CSIA) revealed that both antipatharian species show a high trophic position relative to known mixotrophic and heterotrophic corals. Although both species inhabit the same area and are planktivorous, their carbon signature suggests they have different carbon sources. Our findings imply that antipatharians have a vital role in the trophic dynamics of mesophotic coral ecosystems, a unique habitat whose ecology remains largely unexplored.

Polystyrene nanoplastics impair the photosynthetic capacities of Symbiodiniaceae and promote coral bleaching

Reef-building corals are increasingly threatened by global and regional stresses, which affect the stability of the coral-Symbiodiniaceae association. Among them, plastic pollution has been an ongoing and growing concern. Whereas several studies have highlighted the detrimental impact of microplastics (0.1 μm–5 mm) on corals and their symbiotic dinoflagellate algae, the physiological changes induced by nanoplastic (NP, <0.1 μm) pollution are still poorly known. Long-term experiments (4 weeks) were conducted to investigate the effects of ecologically relevant NP concentrations (0 to 0.5 mg/L of 20 nm polystyrene NPs) on two Symbiodiniaceae in culture [CCMP2467 or Clade A1 and pd44b or Clade F1]. The effects of 0.5 mg/L NPs were also evaluated on Clade A1 living in symbiosis with the coral Stylophora pistillata, to assess the in hospite effects of NPs on coral symbionts. The photosynthetic efficiency of photosystem II, the oxidative status of the Symbiodiniaceae and the coral host, as well as the host-symbiont stability were evaluated at the end of the experiment. Symbiodiniaceae in culture exhibited a significant decrease in the maximal electron transport rate (ETRmax) at NP concentrations as low as 0.005 mg/L, highlighting an impairment of the photosynthetic capacities of the dinoflagellates in presence of nanoplastics. Also, Clade A1 exhibited a significant decrease in its Total Antioxidant Capacity (TAC) and an increase in Lipid Peroxidation (LPO), which evidence oxidative stress and cellular damage. Interestingly, Clade A1 in hospite did not show any signs of oxidative stress, however, the coral host exhibited increased TAC and LPO. Additionally, exposure of S. pistillata to 0.5 mg/L NPs induced significant bleaching (loss of symbionts and photosynthetic pigments). Overall, NPs were detrimental for both the Symbiodiniaceae in culture and the host-symbiont association. In the future, the persistence of reef corals may be severely impacted by the cumulative effects of nanoplastic pollution along with global warming.

Metal accumulation induces oxidative stress and alters carbonic anhydrase activity in corals and symbionts from the largest reef complex in the South Atlantic ocean

We evaluated the influence of metal accumulation on the oxidative status [lipid peroxidation (LPO) and total antioxidant capacity (TAC)] and carbonic anhydrase (CA) activity in host and symbionts of the coral Mussismilia harttii and the hydrocoral Millepora alcicornis collected in Abrolhos Reef Banks (Northeast Brazil), potentially impacted by a major mine dam rupture. Considering metal levels measured in reefs worldwide, Abrolhos corals had higher Fe and Mn levels than expected for preserved offshore reefs. Increasing concentrations of arsenic (As), chromium (Cr) and manganese (Mn) drove inhibition of CA and increased oxidative damage in the hydrocoral M. alcicornis. The impairment of enzymatic activity in the symbiotic algae of M. alcicornis may be related to the oxidative stress condition. The hydrocoral M. alcicornis was more affected by metals than the coral M. harttii, which did not show the expected CA inhibition after metal exposure. Our results suggest that CA activity can be applied as a complementary biomarker to evaluate the physiological impacts of environmental metal contamination in reefs. Also, the metal levels and biochemical biomarkers reported in the present study may provide reference data to monitor the health of reefs impacted by a relevant dam rupture.

Establishment of Host-Algal Endosymbioses: Genetic Response to Symbiont Versus Prey in a Sponge Host.

The freshwater sponge Ephydatia muelleri and its Chlorella-like algal partner is an emerging model for studying animal: algal endosymbiosis. The sponge host is a tractable laboratory organism, and the symbiotic algae are easily cultured. We took advantage of these traits to interrogate questions about mechanisms that govern the establishment of durable intracellular partnerships between hosts and symbionts in facultative symbioses. We modified a classical experimental approach to discern the phagocytotic mechanisms that might be co-opted to permit persistent infections, and identified genes differentially expressed in sponges early in the establishment of endosymbiosis. We exposed algal-free E. muelleri to live native algal symbionts and potential food items (bacteria and native heat-killed algae), and performed RNA-Seq to compare patterns of gene expression among treatments. We found a relatively small but interesting suite of genes that are differentially expressed in the host exposed to live algal symbionts, and a larger number of genes triggered by host exposure to heat-killed algae. The upregulated genes in sponges exposed to live algal symbionts were mostly involved in endocytosis, ion transport, metabolic processes, vesicle-mediated transport, and oxidation–reduction. One of the host genes, an ATP-Binding Cassette transporter that is downregulated in response to live algal symbionts, was further evaluated for its possible role in the establishment of the symbiosis. We discuss the gene expression profiles associated with host responses to living algal cells in the context of conditions necessary for long-term residency within host cells by phototrophic symbionts as well as the genetic responses to sponge phagocytosis and immune-driven pathways.

Feeding of a Scleractinian Coral, Goniopora columna, on Microalgae, Yeast, and Artificial Feed in Captivity.

Simple SummaryCoral aquaculture is an innovative and sustainable aquaculture industry. Coral husbandry can address ecological environment conservation needs and industrial demand for corals. Many previous studies have confirmed that corals also belong to heterotrophic organisms. Heterotrophic feeding is essential for overcoming nutrient deficiency. The preliminary results of this study indicate that Goniopora columna have high levels of proteases, and artificial feeds containing high protein can be used for feeding during aquaculture, which can increase the growth rate. In conclusion, we have initially explored that Goniopora columna will have better growth by feeding artificial PUFA rich in animal protein. In addition, the best feeding time is 6:00–12:00 in the morning, when there is better digestion and absorption. It is hoped that this research will be helpful to the development of coral aquaculture in the future.Nutritional requirements are critical in the process of coral aquaculture. In addition to energy from symbiotic algae, corals obtain sufficient nutrition through heterotrophic feeding. Microalgae and yeast are commonly used as nutritional supplements for many aquaculture organisms. In addition, if artificial feed can match or improve upon the nutritional supplementation provided by microalgae and yeast in the case of G. columna, then feeding this coral would be markedly easier. Hence, this article preliminarily discusses feeds suitable for G. columna. In this study, artificial PUFA rich in animal protein (R), Saccharomyces cerevisiae, Isochrysis galbana tml, and Nannochloropsis oculate were fed to G. columna at quantities of 5% and 10% of body weight. Growth, survival, body composition, and digestive enzymes were assessed. Regarding body composition, the coral’s protein content is higher than that of carbohydrate or fat; thus, evaluating the heterotrophic nutrition of G. columna by using protein absorption is appropriate. The protease content is also high in digestive enzymes. Protein content, protease activity, and specific growth rate were significantly higher in the R group than in other groups. The number of polyps in the groups fed R at 5% and 10% of body weight increased by 40.00 ± 2.43 and 47.33 ± 0.89 number, respectively, significantly greater increases than those achieved in the other groups (p < 0.05). Changes in body composition and digestive enzymes over a 24-h period were compared to determine the optimal feeding time. Protein content and protease activity increased markedly between 6:00 and 12:00. The experimental results suggest that R can improve the activity of G. columna digestive enzymes and their protein and lipid content in body tissue, shorten the cultivation time, and enhance the profitability of coral aquaculture.

Physiological Response of Shallow-Water Hard Coral Acropora digitifera to Heat Stress via Fatty Acid Composition

Heat stress disturbs the mutualistic relationship between the hard corals and the symbiotic algae, which cause coral bleaching. A wide array of biochemical parameters is used to demonstrate the phenomenon. This study exposed a shallow-water hard coral, Acropora digitifera, to a series of elevated temperatures over time while the interaction between Symbiodiniaceae (SD) density, antioxidants activities, fatty acid (FA) composition, and putative FA health indicators was evaluated. Heat stress caused a substantial loss in SD densities, consequently regulated the antioxidant activities and caused significant changes in FA composition. There was a lack of evidence showing A. digitifera experienced oxidative stress; nonetheless, a significant decrease of monounsaturated fatty acid as (MUFA) and polyunsaturated fatty acid (PUFA) during the thermally induced experiment demonstrated that corals utilize their unsaturated FA as a final barrier or as a repair system against oxidative damage once the antioxidant enzyme cannot cope with stress condition. The lower ratio of putative FA health indicators [i.e., n-3 LC:n-6 LC, eicosapentaenoic acid (EPA):arachidonic acid (ARA), and docosahexaenoic acid (DHA):ARA] characterized an unhealthy coral. The loss of SD density was significantly correlated with certain PUFA markers [i.e., linolenic acid (18:3n6), 20:5n3, and 22:6n3] and putative FA health indicator (i.e., n-3 LC:n-6 LC, EPA:ARA, and DHA:ARA). These notably imply that the FA linked with the symbiont can be a potential health indicator for assessing the effect of the environmental stressor on coral. This study also revealed the regulation of FAs during stress conditions, especially when heterotrophic feeding is limited. Future studies on FA profiles toward antagonistic or synergistic effects will offer a better understanding of the nature of this relationship under a harsh climate.

Novel shortcut biological nitrogen removal using activated sludge-biofilm coupled with symbiotic algae

This study evaluated a novel shortcut biological nitrogen removal (BNR) method using activated sludge-biofilm coupled with microalgae in a photo-sequencing batch biofilm reactor (PSBBR) for treating high ammonia nitrogen (NH4+-N) wastewater. Results showed that the symbiotic microalgae supplied oxygen and an additional organic carbon source for bacteria. The consortium achieved high nitrogen removal efficiency (> 90%) and had the potential to lower the operating cost when used for treating biogas slurry (BS). It saved more than 50% of the external carbon source and reduced about 78% of oxygen demand compared with the conventional BNR process. Nitrogen was removed mainly through shortcut nitrification-denitrification (SCND) (> 80%) and biological assimilation (6.8%) in the PSBBR. The algal-bacterial symbiosis improved the species richness but reduced the microbial diversity of the biofilm. Nitrosomonas with an abundance of 19.81% was the dominant ammonia-oxidizing bacteria (AOB) in the biofilm, while Thauera with an abundance of 45.03% played a leading role in denitrification. This study concurrently indicated that algae and activated sludge coupling increased the nitrous oxide (N2O) emission in the shortcut nitrogen removal process.

Effect of Cryopreservation on Proteins from the Ubiquitous Marine Dinoflagellate Breviolum sp. (Family Symbiodiniaceae).

Coral reefs around the world are exposed to thermal stress from climate change, disrupting the delicate symbiosis between the coral host and its symbionts. Cryopreservation is an indispensable tool for the preservation of species, as well as the establishment of a gene bank. However, the development of cryopreservation techniques for application to symbiotic algae is limited, in addition to the scarceness of related studies on the molecular level impacts post-thawing. Hence, it is essential to set up a suitable freezing protocol for coral symbionts, as well as to analyze its cryo-injury at the molecular level. The objective of this study was to develop a suitable protocol for the coral symbiont Breviolum subjected to two-step freezing. The thawed Breviolum were then cultured for 3, 7, 14, and 28 days before they were analyzed by Western blot for protein expression, light-harvesting protein (LHP), and red fluorescent protein (RFP) and tested by adenosine triphosphate bioassay for cell viability. The results showed the highest cell viability for thawed Breviolum that was treated with 2 M propylene glycol (PG) and 2 M methanol (MeOH) and equilibrated with both cryoprotectants for 30 min and 20 min. Both treatment groups demonstrated a significant increase in cell population after 28 days of culture post-thawing, especially for the MeOH treatment group, whose growth rate was twice of the PG treatment group. Regarding protein expression, the total amounts of each type of protein were significantly affected by cryopreservation. After 28 days of culture, the protein expression for the MeOH treatment group showed no significant difference to that of the control group, whereas the protein expression for the PG treatment group showed a significant difference. Breviolum that were frozen with MeOH recovered faster upon thawing than those frozen with PG. LHP was positively and RFP was negatively correlated with Symbiodiniaceae viability and so could serve as health-informing biomarkers. This work represents the first time to document it in Symbiodiniaceae, and this study established a suitable protocol for the cryopreservation of Breviolum and further refined the current understanding of the impact of low temperature on its protein expression. By gaining further understanding of the use of cryopreservation as a way to conserve Symbiodiniaceae, we hope to make an effort in the remediation and conservation of the coral reef ecosystem and provide additional methods to rescue coral reefs.

Impact of Seawater Temperature on Coral Reefs in the Context of Climate Change. A Case Study of Cu Lao Cham – Hoi An Biosphere Reserve

Coral reefs are a natural habitat for many species, as well as being of high economic and touristic significance. However, they represent an extremely sensitive ecosystem with a narrow ecological limit: prolonged high temperatures can lead to bleaching, in which corals expel their symbiotic algae and eventually corals will degrade and die. Based on climate change projections from the Blue Communities regional model, using linear regression, exponential regression, polynomial regression, we found that by the decades 2041–2050 and 2051–2060, whether with RCP 4.5 or RCP 8.5, the environmental temperature will change beyond the coral capacity threshold. Of particular concern is RCP 8.5, where the number of weeks per decade in which SST exceeds the threshold of coral reef bleaching is up to 55, compared to 0 at the beginning of the century. As well, the El Niño phenomenon often heats up waters to abnormally high temperatures in Cu Lao Cham and, it is projected to rise even further. Consequently, the combination of climate change and El Niño will cause abnormal increases in the seawater environment beyond the coral resistance threshold, leading to degradation of this internationally important site. Decisive and practical action must be taken to deal with climate change in this part of the world.

Contrasting heat stress response patterns of coral holobionts across the Red Sea suggest distinct mechanisms of thermal tolerance.

Corals from the northern Red Sea, in particular the Gulf of Aqaba (GoA), have exceptionally high bleaching thresholds approaching >5℃ above their maximum monthly mean (MMM) temperatures. These elevated thresholds are thought to be due to historical selection, as corals passed through the warmer Southern Red Sea during recolonization from the Arabian Sea. To test this hypothesis, we determined thermal tolerance thresholds of GoA versus central Red Sea (CRS) Stylophora pistillata corals using multi-temperature acute thermal stress assays to determine thermal thresholds. Relative thermal thresholds of GoA and CRS corals were indeed similar and exceptionally high (~7℃ above MMM). However, absolute thermal thresholds of CRS corals were on average 3℃ above those of GoA corals. To explore the molecular underpinnings, we determined gene expression and microbiome response of the coral holobiont. Transcriptomic responses differed markedly, with a strong response to the thermal stress in GoA corals and their symbiotic algae versus a remarkably muted response in CRS colonies. Concomitant to this, coral and algal genes showed temperature-induced expression in GoA corals, while exhibiting fixed high expression (front-loading) in CRS corals. Bacterial community composition of GoA corals changed dramatically under heat stress, whereas CRS corals displayed stable assemblages. We interpret the response of GoA corals as that of a resilient population approaching a tipping point in contrast to a pattern of consistently elevated thermal resistance in CRS corals that cannot further attune. Such response differences suggest distinct thermal tolerance mechanisms that may affect the response of coral populations to ocean warming.

The skeleton of Balanophyllia coral species suggests adaptive traits linked to the onset of mixotrophy

The diversity in the skeletal features of coral species is an outcome of their evolution, distribution and habitat. Here, we explored, from macro- to nano-scale, the skeletal structural and compositional characteristics of three coral species belonging to the genus Balanophyllia having different trophic strategies. The goal is to address whether the onset of mixotrophy influenced the skeletal features of B. elegans, B. regia, and B. europaea. The macroscale data suggest that the presence of symbiotic algae in B. europaea can lead to a surplus of energy input that increases its growth rate and skeletal bulk density, leading to larger and denser corals compared to the azooxanthellate ones, B. regia and B. elegans. The symbiosis would also explain the higher intra-skeletal organic matrix (OM) content, which is constituted by macromolecules promoting the calcification, in B. europaea compared to the azooxanthellate species. The characterization of the soluble OM also revealed differences between B. europaea and the azooxanthellate species, which may be linked to diverse macromolecular machineries responsible for skeletal biosynthesis and final morphology. Differently, the crystallographic features were homogenous among species, suggesting that the basic building blocks of skeletons remained a conserved trait in these related species, regardless of the trophic strategy. These results show changes in skeletal phenotype that could be triggered by the onset of mixotrophy, as a consequence of the symbiotic association, displaying remarkable plasticity of coral skeletons which repeatedly allowed this coral group to adapt to a range of changing environments throughout its geological history.

The Endosymbiotic Coral Algae Symbiodiniaceae Are Sensitive to a Sensory Pollutant: Artificial Light at Night, ALAN.

Artificial Light at Night, ALAN, is a major emerging issue in biodiversity conservation, which can negatively impact both terrestrial and marine environments. Therefore, it should be taken into serious consideration in strategic planning for urban development. While the lion’s share of research has dealt with terrestrial organisms, only a handful of studies have focused on the marine milieu. To determine if ALAN impacts the coral reef symbiotic algae, that are fundamental for sustainable coral reefs, we conducted a short experiment over a period of one-month by illuminating isolated Symbiodiniaceae cell cultures from the genera Cladocopium (formerly Clade C) and Durusdinium (formerly Clade D) with LED light. Cell cultures were exposed nightly to ALAN levels of 0.15 μmol quanta m–2 s–1 (∼4–5 lux) with three light spectra: blue, yellow and white. Our findings showed that even in very low levels of light at night, the photo-physiology of the algae’s Electron Transport Rate (ETR), Non-Photochemical Quenching, (NPQ), total chlorophyll, and meiotic index presented significantly lower values under ALAN, primarily, but not exclusively, in Cladocopium cell cultures. The findings also showed that diverse Symbiodiniaceae types have different photo-physiology and photosynthesis performances under ALAN. We believe that our results sound an alarm for the probable detrimental effects of an increasing sensory pollutant, ALAN, on the eco-physiology of symbiotic corals. The results of this study point to the potential effects of ALAN on other organisms in marine ecosystem such as fish, zooplankton, and phytoplankton in which their biorhythms is entrained by natural light and dark cycles.

The ciliate Paramecium bursaria allows budding of symbiotic Chlorella variabilis cells singly from the digestive vacuole membrane into the cytoplasm during algal reinfection.

The ciliate Paramecium bursaria harbors several hundred symbiotic Chlorella spp. cells in the cytoplasm. Algal re-endosymbiosis can be artificially induced using alga-removed P. bursaria. During algal re-endosymbiosis, algae ingested into the host digestive vacuoles (DVs) avoid digestion by the host lysosomal enzymes and then escape into the cytoplasm by budding off of the DV membrane. The budded alga-enclosing DV membrane then differentiates into the symbiosome or perialgal vacuole (PV) membrane and is localized beneath the host cell cortex. In this study, we determined whether the PV membrane has the ability to recognize the symbiotic alga singly by eliminating other small microspheres in the same DV. To clarify the accuracy of the budding process, we mixed fluorescent-labeled microspheres of diameter 0.20µm with isolated symbiotic algae during algal re-endosymbiosis. No fluorescence was observed from the PV membrane, as expected, and the budding DVs that enclosed both undigested and digested algae. Additionally, the algal re-endosymbiosis rate was significantly reduced in the presence of microspheres. These observations showed that the host P. bursaria allowed budding of the algae singly from the membranes of DVs without microspheres and this process required close contact between the DV membrane and the algal cell wall.

Genetic Diversity of Symbiotic Green Algae of Paramecium bursaria Syngens Originating from Distant Geographical Locations.

Paramecium bursaria (Ehrenberg 1831) is a ciliate species living in a symbiotic relationship with green algae. The aim of the study was to identify green algal symbionts of P. bursaria originating from distant geographical locations and to answer the question of whether the occurrence of endosymbiont taxa was correlated with a specific ciliate syngen (sexually separated sibling group). In a comparative analysis, we investigated 43 P. bursaria symbiont strains based on molecular features. Three DNA fragments were sequenced: two from the nuclear genomes—a fragment of the ITS1-5.8S rDNA-ITS2 region and a fragment of the gene encoding large subunit ribosomal RNA (28S rDNA), as well as a fragment of the plastid genome comprising the 3′rpl36-5′infA genes. The analysis of two ribosomal sequences showed the presence of 29 haplotypes (haplotype diversity Hd = 0.98736 for ITS1-5.8S rDNA-ITS2 and Hd = 0.908 for 28S rDNA) in the former two regions, and 36 haplotypes in the 3′rpl36-5′infA gene fragment (Hd = 0.984). The following symbiotic strains were identified: Chlorella vulgaris, Chlorella variabilis, Chlorella sorokiniana and Micractinium conductrix. We rejected the hypotheses concerning (i) the correlation between P. bursaria syngen and symbiotic species, and (ii) the relationship between symbiotic species and geographic distribution.

Unique environmental Symbiodiniaceae diversity at an isolated island in the northwestern Pacific

Dinoflagellates in the family Symbiodiniaceae are intensively investigated as algal symbionts of corals and other invertebrates, underpinning coral reef ecosystems as primary producers. Diversity, including regional diversification, of free-living communities is less studied. In this study, an environmental Symbiodiniaceae community at an isolated island, Okinotori Island, Japan, was investigated to determine whether the community is endemic or common with other locations near continents and major ocean currents. Symbiotic algae in common corals at the island were the same type as those of the corals from other Japanese waters. In the environmental samples, genera Symbiodinium (formerly clade A), Cladocopium (clade C), Durusdinium (clade D), and clades F (including Freudenthalidium), G, and I, were identified through analysis of internal transcribed spacer region 2 of nuclear ribosomal RNA gene (ITS2) sequences. Interestingly, some sequences found were genetically different from those of previously reported genera/clades. These unknown sequences were genetically included in the Symbiodiniaceae linage, but they were differentiated from the previously known nine clades. The sequences formed a cluster in the phylogenetic tree based on 28S nrDNA. These sequences were thus considered members of a novel clade in the family (clade J). In total, 120 kinds of ITS2 sequences were produced; while 10 were identical to previously reported sequences, the majority were highly divergent. These genetically unique Symbiodiniaceae types, including novel clade J, may have evolved in isolation and reflect the environmental characteristics of the Okinotori Island.