Choosing the Right Life Partner: Ecological Drivers of Lichen Symbiosis

Lichens are an iconic example of symbiotic systems whose ecology is shaped by the requirements of the symbionts. Previous studies suggest that fungal (mycobionts) as well as photosynthesizing (phycobionts or cyanobionts) partners have a specific range of acceptable symbionts that can be chosen according to specific environmental conditions. This study aimed to investigate the effects of climatic conditions and mycobiont identity on phycobiont distribution within the lichen genera Stereocaulon, Cladonia, and Lepraria. The study area comprised the Canary Islands, Madeira, Sicily, and the Aeolian Islands, spanning a wide range of climatic conditions. These islands are known for their unique and diverse fauna and flora; however, lichen phycobionts have remained unstudied in most of these areas. In total, we genetically analyzed 339 lichen samples. The phycobiont pool differed significantly from that outside the studied area. Asterochloris mediterranea was identified as the most abundant phycobiont. However, its distribution was limited by climatic constraints. Other species of Asterochloris and representatives of the genera Chloroidium, Vulcanochloris, and Myrmecia were also recovered as phycobionts. The selection of symbiotic partners from the local phycobiont pool was driven by mycobiont specificity (i.e., the taxonomic range of acceptable partners) and the environmental conditions, mainly temperature. Interestingly, the dominant fungal species responded differently in their selection of algal symbionts along the environmental gradients. Cladonia rangiformis associated with its phycobiont A. mediterranea in a broader range of temperatures than Stereocaulon azoreum, which favors other Asterochloris species along most of the temperature gradient. Stereocaulon vesuvianum associated with Chloroidium spp., which also differed in their temperature optima. Finally, we described Stereocaulon canariense as a new endemic species ecologically distinct from the other Stereocaulon species on the Canary Islands.

Heat stress reduces the contribution of diazotrophs to coral holobiont nitrogen cycling

Efficient nutrient cycling in the coral-algal symbiosis requires constant but limited nitrogen availability. Coral-associated diazotrophs, i.e., prokaryotes capable of fixing dinitrogen, may thus support productivity in a stable coral-algal symbiosis but could contribute to its breakdown when overstimulated. However, the effects of environmental conditions on diazotroph communities and their interaction with other members of the coral holobiont remain poorly understood. Here we assessed the effects of heat stress on diazotroph diversity and their contribution to holobiont nutrient cycling in the reef-building coral Stylophora pistillata from the central Red Sea. In a stable symbiotic state, we found that nitrogen fixation by coral-associated diazotrophs constitutes a source of nitrogen to the algal symbionts. Heat stress caused an increase in nitrogen fixation concomitant with a change in diazotroph communities. Yet, this additional fixed nitrogen was not assimilated by the coral tissue or the algal symbionts. We conclude that although diazotrophs may support coral holobiont functioning under low nitrogen availability, altered nutrient cycling during heat stress abates the dependence of the coral host and its algal symbionts on diazotroph-derived nitrogen. Consequently, the role of nitrogen fixation in the coral holobiont is strongly dependent on its nutritional status and varies dynamically with environmental conditions.

Factors Limiting the Range Extension of Corals into High-Latitude Reef Regions

Reef-building corals show a marked decrease in total species richness from the tropics to high latitude regions. Several hypotheses have been proposed to account for this pattern in the context of abiotic and biotic factors, including temperature thresholds, light limitation, aragonite saturation, nutrient or sediment loads, larval dispersal constraints, competition with macro-algae or other invertebrates, and availability of suitable settlement cues or micro-algal symbionts. Surprisingly, there is a paucity of data supporting several of these hypotheses. Given the immense pressures faced by corals in the Anthropocene, it is critical to understand the factors limiting their distribution in order to predict potential range expansions and the role that high latitude reefs can play as refuges from climate change. This review examines these factors and outlines critical research areas to address knowledge gaps in our understanding of light/temperature interactions, coral-Symbiodiniaceae associations, settlement cues, and competition in high latitude reefs.

Effect of species, size, and chimerism on the susceptibility of Caribbean brain coral recruits to stony coral tissue loss disease (SCTLD)

Stony coral tissue loss disease (SCTLD) has devastated coral populations along Florida′s Coral Reef and beyond. Although widespread infection and mortality of adult colonies have been documented, no studies have yet investigated the susceptibility of recruits to this disease. Here, we exposed eight-month-old Diploria labyrinthiformis recruits and four-month-old Colpophyllia natans recruits to two sequential doses of SCTLD in the laboratory to track infection and assess potential resilience. Both species began to develop lesions as early as 48 h after exposure began. During the first dose, 59.0% of C. natans recruits lost all tissue (died) within two to eight days of developing lesions, whereas D. labyrinthiformis recruits experienced significantly slower rates of tissue loss and minimal eventual mortality. In C. natans, larger recruits and those fused into groups of multiple genets (chimeras) exhibited the highest survivorship. In contrast, smaller and/or single (ungrouped) recruits had the lowest survivorship (9.9 - 26.5%). After 20 days, a second SCTLD dose was delivered to further test resistance in remaining recruits, and all recruits of both species succumbed within 6 days. Although no recruits showed absolute resistance to SCTLD following repeated exposures, our results provide evidence that interactions between species, size, and chimerism can impact relative resistance. This study represents the first report of SCTLD in Caribbean coral recruits and carries implications for natural species recovery and reef restoration efforts. Additional research on the susceptibility of coral juveniles to SCTLD is urgently needed, to include different species, locations, parents, and algal symbionts, with the goal of assessing relative susceptibility and identifying potential sources of resilience for this critical life history stage.

Sedimentary Nitrogen Uptake & Assimilation in the Temperate Zooxanthellate Anemone Anthopleura aureoradiata

<p>This study investigated the potential for, and efficiency of, particulate nitrogen uptake from the sediment and subsequent consequences of this on the nutrient status of endosymbiotic dinoflagellates (zooxanthellae) in the temperate zooxanthellate anemone Anthopleura aureoradiata. Sediment was collected from a mudflat and labelled with (15NH4)2SO4 before being provided to A. aureoradiata at low (5 g dry weight) and high sediment (20 g dry weight) loads for 6 hours. While no discernible change in the isotopic content of the sediment could be detected, analysis of the host and algal symbionts revealed that 15N had been taken up. Uptake by the host was similar at both high and low sediment loads, but the algal symbionts acquired more nitrogen at the lower load (1.13 versus 0.93 atom % 15N in the low and high loads, respectively). Evaluation of this particulate nitrogen uptake from the sediment was further examined by measuring the nitrogen status of the zooxanthellae. This was determined by measuring the extent to which ammonium (40 muM NH+4) enhanced the rate of zooxanthellar dark carbon fixation above that seen in filtered seawater (FSW) alone; the enhancement ratio was expressed as [dark NH+4 rate/dark FSW rate]. VD'/VL, a further index of nitrogen status, was also calculated where VD' = [dark NH+4 rate - dark FSW rate] and VL = rate of carbon fixation in the light. When anemones were starved for 2-8 weeks, zooxanthellar nitrogen deficiency became apparent at greater than or equal to 4 weeks, with NH+4/FSW and VD'/VL averaging up to 2.90 and 0.11, respectively. In comparison, when anemones were fed 5 times per week for 8 weeks the addition of ammonium had little effect, indicating nitrogen sufficiency; NH+4/FSW and VD'/VL values were 1.03 and -1.0 x 10-3, respectively. The nitrogen status of zooxanthellae from anemones starved and incubated with and without sediment was examined with no apparent difference between sediment and no sediment treatments; zooxanthellar nitrogen deficiency became apparent at greater than or equal to 4 weeks in both treatments, with NH+4/FSW and VD'/VL averaging up to 3.73 and 0.17 for the sediment treatment and 2.74 and 0.15 for the no sediment treatment, respectively. The nitrogen status of zooxanthellae from anemones found on a mudflat (Pauatahanui Inlet) and a rocky intertidal site (Kau Bay) was different. Zooxanthellae from mudflat anemones were nitrogen sufficient with NH+4/FSW and VD'/VL values averaging up to 1.26 and -6.0 x 10-3, respectively. Nitrogen deficient zooxanthellae were present in anemones from the rocky intertidal. Anemones from tide pools in the upper littoral zone had NH+4/FSW and VD'/VL values of 2.99 and 0.11, respectively, while anemones from the mid littoral zone had NH+4/FSW and VD'/VL of 2.90 and 0.13, respectively; there was no significant difference in nitrogen status between zooxanthellae from high shore tide pool anemones and aerially exposed mid-littoral anemones. These results suggest that while particulate nitrogen can be taken up from the sediment by this species, dissolved inorganic nitrogen such as ammonium in the seawater, and especially the interstitial water surrounding infaunal anemones on mudflats, may be a more important source of nitrogen in the field.</p>

Sedimentary Nitrogen Uptake & Assimilation in the Temperate Zooxanthellate Anemone Anthopleura aureoradiata

<p>This study investigated the potential for, and efficiency of, particulate nitrogen uptake from the sediment and subsequent consequences of this on the nutrient status of endosymbiotic dinoflagellates (zooxanthellae) in the temperate zooxanthellate anemone Anthopleura aureoradiata. Sediment was collected from a mudflat and labelled with (15NH4)2SO4 before being provided to A. aureoradiata at low (5 g dry weight) and high sediment (20 g dry weight) loads for 6 hours. While no discernible change in the isotopic content of the sediment could be detected, analysis of the host and algal symbionts revealed that 15N had been taken up. Uptake by the host was similar at both high and low sediment loads, but the algal symbionts acquired more nitrogen at the lower load (1.13 versus 0.93 atom % 15N in the low and high loads, respectively). Evaluation of this particulate nitrogen uptake from the sediment was further examined by measuring the nitrogen status of the zooxanthellae. This was determined by measuring the extent to which ammonium (40 muM NH+4) enhanced the rate of zooxanthellar dark carbon fixation above that seen in filtered seawater (FSW) alone; the enhancement ratio was expressed as [dark NH+4 rate/dark FSW rate]. VD'/VL, a further index of nitrogen status, was also calculated where VD' = [dark NH+4 rate - dark FSW rate] and VL = rate of carbon fixation in the light. When anemones were starved for 2-8 weeks, zooxanthellar nitrogen deficiency became apparent at greater than or equal to 4 weeks, with NH+4/FSW and VD'/VL averaging up to 2.90 and 0.11, respectively. In comparison, when anemones were fed 5 times per week for 8 weeks the addition of ammonium had little effect, indicating nitrogen sufficiency; NH+4/FSW and VD'/VL values were 1.03 and -1.0 x 10-3, respectively. The nitrogen status of zooxanthellae from anemones starved and incubated with and without sediment was examined with no apparent difference between sediment and no sediment treatments; zooxanthellar nitrogen deficiency became apparent at greater than or equal to 4 weeks in both treatments, with NH+4/FSW and VD'/VL averaging up to 3.73 and 0.17 for the sediment treatment and 2.74 and 0.15 for the no sediment treatment, respectively. The nitrogen status of zooxanthellae from anemones found on a mudflat (Pauatahanui Inlet) and a rocky intertidal site (Kau Bay) was different. Zooxanthellae from mudflat anemones were nitrogen sufficient with NH+4/FSW and VD'/VL values averaging up to 1.26 and -6.0 x 10-3, respectively. Nitrogen deficient zooxanthellae were present in anemones from the rocky intertidal. Anemones from tide pools in the upper littoral zone had NH+4/FSW and VD'/VL values of 2.99 and 0.11, respectively, while anemones from the mid littoral zone had NH+4/FSW and VD'/VL of 2.90 and 0.13, respectively; there was no significant difference in nitrogen status between zooxanthellae from high shore tide pool anemones and aerially exposed mid-littoral anemones. These results suggest that while particulate nitrogen can be taken up from the sediment by this species, dissolved inorganic nitrogen such as ammonium in the seawater, and especially the interstitial water surrounding infaunal anemones on mudflats, may be a more important source of nitrogen in the field.</p>

Insights into early stages of the establishment of host:algal endosymbioses: Genetic responses to live versus heat-killed algae and bacterial prey in a sponge host

The freshwater sponge Ephydatia muelleri and its Chlorella-like green algal partner is an emerging model for studying animal:algal endosymbiosis. The sponge host is a highly tractable laboratory organism, and the symbiotic algae are easily cultured. We took advantage of these traits to experimentally interrogate fundamental questions about cellular 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 E. muelleri sponges early in the establishment of endosymbiosis. We exposed algal-free E. muelleri to live native algal symbionts, native heat-killed algae, and bacteria, and then performed RNASeq so we could compare patterns of gene expression in each treatment. We contrasted differential gene expression patterns between potential food items (bacteria and heat-killed algae) and the live native Chlorella-like symbiont. We found a relatively small but interesting suite of genes that are uniquely and differentially expressed in the host exposed to live algal symbionts, and a larger number of genes uniquely triggered by host exposure to heat-killed algae. One of the host genes, an ABC transporter that is downregulated in response to live algal symbionts, was further evaluated for its possible role in establishment of the algal 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.

Differential Symbiodiniaceae Association With Coral and Coral-Eroding Sponge in a Bleaching Impacted Marginal Coral Reef Environment

Marginal reefs are known for severe stress-inducible perturbations such as high sedimentation, eutrophication, ocean warming, and acidification from anthropogenic climate change. The corals striving in such stressful environments develop physiological adaptations induced by differential genomic expressions or association with thermal stress-tolerant algal symbionts (Symbiodiniaceae). Despite such adaptations, corals are threatened by other space competitors such as algae and sponges. Coral-eroding sponges belonging to the Cliona viridis complex are one such space competitors that also associate with Symbiodiniaceae algal photosymbiont. The diversity of Symbiodiniaceae associates with the coral and sponge from the same ecosystems is scarcely known. In the present study, Symbiodiniaceae community structure in the coral Turbinaria mesenterina, a newly described coral-eroding sponge Cliona thomasi, and their surrounding seawater was determined from the nearshore marginal reef along the central west coast of India. The results revealed a significantly higher relative abundance of Durusdinium and Gerakladium than Symbiodinium and Cladocopium in the seawater. Interestingly, both investigated host species showed differential Symbiodiniaceae association with significantly higher abundance of Durusdinium in coral and Gerakladium in sponge. The beta diversity analysis by Permutational multivariate analysis of variance (PERMANOVA) confirmed significant differences in Symbiodiniaceae profiles between sponge and coral. Durusdinium and Gerakladium are thermotolerant genera known to associate with different hosts in suboptimal conditions. Our field surveys suggested the bleaching resistance of the coral T. mesenterina despite the fact that the sea surface temperature reached the coral thermal threshold of 31°C during different periods of the years 2015, 2016, 2017, 2018, and 2019. Therefore, the thermal tolerance of the investigated coral and sponge species may be attributed to their respective thermotolerant photosymbiont associations. Furthermore, the results also indicated the host-specific photosymbiont selection from the local environment. Although these observations provide valuable biological insight, more research is needed to understand the tripartite association of sponge-coral-symbiont together to evaluate the competitive fitness of holobionts.