Temperate Anemone Research Articles

Genetic structure in the endosymbiont Breviolum 'muscatinei' is correlated with geographical location, environment and host species.

Corals and cnidarians form symbioses with dinoflagellates across a wide range of habitats from the tropics to temperate zones. Notably, these partnerships create the foundation of coral reef ecosystems and are at risk of breaking down due to climate change. This symbiosis couples the fitness of the partners, where adaptations in one species can benefit the holobiont. However, the scales over which each partner can match their current—and future—environment are largely unknown. We investigated population genetic patterns of temperate anemones (Anthopleura spp.) and their endosymbiont Breviolum ‘muscatinei’, across an extensive geographical range to identify the spatial scales over which local adaptation is possible. Similar to previously published results, two solitary host species exhibited isolation by distance across hundreds of kilometres. However, symbionts exhibited genetic structure across multiple spatial scales, from geographical location to depth in the intertidal zone, and host species, suggesting that symbiont populations are more likely than their hosts to adaptively mitigate the impact of increasing temperatures.

Microbiome Composition and Diversity of the Ice-Dwelling Sea Anemone, Edwardsiella andrillae.

Edwardsiella andrillae is a sea anemone (Cnidaria: Anthozoa: Actiniaria) only known to live embedded in the ice at the seawater interface on the underside of the Ross Ice Shelf, Antarctica. Although the anatomy and morphological characteristics of E. andrillae have been described, the adaptations of this species to the under-ice ecosystem have yet to be examined. One feature that may be important to the physiology and ecology of E. andrillae is its microbiome, which may play a role in health and survival, as has been deduced in other metazoans, including anthozoans. Here we describe the microbiome of five specimens of E. andrillae, compare the diversity we recovered to that known for temperate anemones and another Antarctic cnidarian, and consider the phylogenetic and functional implications of microbial diversity for these animals. The E. andrillae microbiome was relatively low in diversity, with seven phyla detected, yet included substantial phylogenetic novelty. Among the five anemones investigated, the distribution of microbial taxa varied; this trait appears to be shared by many anthozoans. Most importantly, specimens either appeared to be dominated by Proteobacteria-affiliated members or by deeply branching Tenericute sequences. There were few closely related sequence types that were common to temperate and Antarctic sea anemone microbiomes, the exception being an Acinetobacter-related representative. Similar observations were made between microbes associated with E. andrillae and an Antarctic soft coral; however, there were several closely-related, low abundance Gammaproteobacteria in both Antarctic microbiomes, particularly from the soft coral, that are also commonly detected in Southern Ocean seawater. Although this preliminary study leaves open many questions concerning microbiome diversity and its role in host ecology, we identify major lineages of microbes (e.g., diverse deep-branching Alphaproteobacteria, Epsilonproteobacteria, and divergent Tenericutes affiliates) that may play critical roles, and we highlight the current understanding and the need for future studies of sea anemone-microbiome relationships.

De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes.

Many nonmodel species exemplify important biological questions but lack the sequence resources required to study the genes and genomic regions underlying traits of interest. Reef-building corals are famously sensitive to rising seawater temperatures, motivating ongoing research into their stress responses and long-term prospects in a changing climate. A comprehensive understanding of these processes will require extending beyond the sequenced coral genome (Acropora digitifera) to encompass diverse coral species and related anthozoans. Toward that end, we have assembled and annotated reference transcriptomes to develop catalogs of gene sequences for three scleractinian corals (Fungia scutaria, Montastraea cavernosa, Seriatopora hystrix) and a temperate anemone (Anthopleura elegantissima). High-throughput sequencing of cDNA libraries produced ∼20–30 million reads per sample, and de novo assembly of these reads produced ∼75,000–110,000 transcripts from each sample with size distributions (mean ∼1.4 kb, N50 ∼2 kb), comparable to the distribution of gene models from the coral genome (mean ∼1.7 kb, N50 ∼2.2 kb). Each assembly includes matches for more than half the gene models from A. digitifera (54–67%) and many reasonably complete transcripts (∼5300–6700) spanning nearly the entire gene (ortholog hit ratios ≥0.75). The catalogs of gene sequences developed in this study made it possible to identify hundreds to thousands of orthologs across diverse scleractinian species and related taxa. We used these sequences for phylogenetic inference, recovering known relationships and demonstrating superior performance over phylogenetic trees constructed using single mitochondrial loci. The resources developed in this study provide gene sequences and genetic markers for several anthozoan species. To enhance the utility of these resources for the research community, we developed searchable databases enabling researchers to rapidly recover sequences for genes of interest. Our analysis of de novo assembly quality highlights metrics that we expect will be useful for evaluating the relative quality of other de novo transcriptome assemblies. The identification of orthologous sequences and phylogenetic reconstruction demonstrates the feasibility of these methods for clarifying the substantial uncertainties in the existing scleractinian phylogeny.

Distribution Patterns of Zoochlorellae and Zooxanthellae Hosted by Two Pacific Northeast Anemones, Anthopleura elegantissima and A. xanthogrammica

This study investigated patterns in the relative abundance of two photosynthetic algal symbionts, zoochlorellae (ZC) and zooxanthellae (ZX), hosted by two temperate anemones, Anthopleura elegantissima and A. xanthogrammica. Previous studies have documented varying proportions of each symbiont along environmental gradients, presumably determined by their respective physiological capabilities. To test for differences in the algal type between the two host species, we sampled anemone tissues (tentacle or tentacle and body column) of similarly sized polyps that were located close together in multiple habitats: tidepools, crevices, underneath rock ledges, and along natural light gradients in caves. The ZC-A. elegantissima symbiosis was rare on the west coast of Vancouver Island, British Columbia, Canada. Even in low-irradiance habitats, ZC were the dominant algae hosted by A. xanthogrammica, while nearby A. elegantissima hosted ZX or was algae-free. As a first step in determining whether symbiont growth rates differed between the two host species, we quantified mitotic index (MI), the percentage of cells with division furrows, under artificial light and in the field by simultaneously sampling tentacles from both species. MI was more stable in A. elegantissima: the MI of ZX isolated from the tentacles of A. xanthogrammica was slightly higher at a light level of 80 micromol quanta m(-2) s(-1) than it was for ZX from A. elegantissima (respectively, 7.3 vs. 6.2) and relatively lower at 40 micromol quanta m(-2) s(-1) (3.9 vs. 5.6). Our data indicate host-specific differences in symbiont distributions and MI when extrinsic physical parameters were similar.

Is dimethylsulfoniopropionate (DMSP) produced by the symbionts or the host in an anemone–zooxanthella symbiosis?

Many groups of tropical cnidarians including scleractinian corals, octocorals, corallimorphs, and anemones contain the tertiary sulfonium compound dimethylsulfoniopropionate (DMSP). It is not known if the compound is synthesized by the animals, their microalgal symbionts, or derived through their diet. We determined the source of the DMSP in several species of tropical and temperate anemones using three approaches: (1) conducting comparative measurements of DMSP in aposymbiotic and zooxanthellate anemones of three species that harbor zooxanthellae, and similar measurements in one species that can harbor both zooxanthellae and zoochlorellae, (2) manipulating the presence or absence of zooxanthellae by inoculating juvenile aposymbiotic anemones (Aiptasia pallida) with their symbiont, Symbiodinium bermudense, and (3) manipulating the numbers of S. bermudense by growing aposymbiotic and zooxanthellate A. pallida in the light and the dark. DMSP was present in zooxanthellate anemones in concentrations of 3.4–15 μmol g−1 fresh mass (FM). In aposymbiotic Aiptasia spp. and Anthopleura elegantissima that lacked large numbers of zooxanthellae, concentrations ranged from being undetectable to 0.43 μmol g−1 FM. When aposymbiotic A. pallida were inoculated with zooxanthellae, concentrations of DMSP were an average of 4.24 μmol g−1 FM after 5 weeks; DMSP was undetectable in uninoculated control animals. Aposymbiotic anemones maintained in the light or the dark for 6 weeks contained no DMSP or zooxanthellae. Zooxanthellate anemones in the light contained five times as many zooxanthellae and approximately 7.5 times as much DMSP as zooxanthellate anemones maintained in the dark. Taken together, these data show that the zooxanthellae are the sole source of DMSP in A. pallida. The trends in DMSP concentrations in other species of zooxanthellate anemones suggest that this phenomenon is not limited to A. pallida but may be more generally true for other anemones or even other cnidarians hosting species of Symbiodinium.

A comparative analysis of the photobiology of zooxanthellae and zoochlorellae symbiotic with the temperate clonal anemone Anthopleura elegantissima (Brandt)

The temperate anemone Anthopleura elegantissima hosts two phylogenetically different symbiotic microalgae, a dinoflagellate Symbiodinium (zooxanthellae, ZX) and a chlorophyte (zoochlorellae, ZC), throughout certain regions of its latitudinal range. Because of the broad intertidal and geographic range of this anemone, we examined the role of irradiance to ascertain which specific symbiotic parameters are affected and whether light intensity governs the observed distributions of natural populations of ZX and ZC. Irradiance appears to be a key factor in regulating both the photophysiology and metabolism of this alga–cnidarian association. Regardless of light intensity, algal densities remained stable for anemones harboring ZX or ZC, whereas the mitotic indices of ZX and ZC both varied directly with light intensity. The chlorophyll content of ZX remained fairly constant regardless of irradiance; in contrast, ZC chlorophyll content was inversely proportional to light intensity. Regardless of irradiance, the carotenoid content of both symbionts was constant; however, ZX carotenoid levels were higher than those of ZC. Net photosynthesis was directly related to light intensity for both algal symbionts and ZX photosynthetic rates were consistently higher than those of ZC. Similarly, the potential carbon contribution of ZX and ZC to animal respiration (CZAR) displayed a direct relationship with light intensity, peaking at 800 µmol·m–2·s–1, then subsequently declined. Lower ZX growth rates, coupled with higher photosynthetic rates and higher CZAR estimates, compared to ZC, suggest that the ZX should be the dominant symbiont as light intensity increases; this may explain the high densities of anemones in the field containing ZX where the levels of irradiance are naturally high. These results support the interpretation that irradiance is a significant environmental parameter that dictates the microhabitat and latitudinal distribution of the two symbiotic algal taxa. This is the second in a series of papers examining the physical parameters that influence the distribution of ZX- and ZC-bearing A. elegantissima.

Temperate and tropical algal‐sea anemone symbioses

Abstract. In this review, we seek to develop new insights about the nature of algal‐sea anemone symbioses by comparing such associations in temperate and tropical seas. Temperate seas undergo pronounced seasonal cycles in irradiance, temperature, and nutrients, while high irradiance, high temperature, and low nutrients are seasonally far less variable in tropical seas. We compare the nature of symbiosis between sea anemones (= actinians) and zooxanthellae (Symbiodinium spp.) in both regions to test tropical paradigms against temperate examples and to identify directions for future research. Although fewer anemone species are symbiotic in temperate regions, they are locally dominant and ecologically important members of the benthic community compared to the tropics.Zooxanthella densities tend to be lower in temperate anemones, but data are limited to a few species in both temperate and tropical seas. Zooxanthella densities are far more stable over time in temperate anemones than in tropical anemones, suggesting that temperate symbioses are more resistant to fluctuations in environmental parameters such as irradiance and temperature. Light‐saturated photosynthetic rates of temperate and tropical zooxanthellae are similar, but temperate anemone hosts receive severely reduced carbon supplies from zooxanthellae during winter months when light is reduced. Symbiont transmission modes and specificity do not show any trends among anemones in tropical vs. temperate seas.Our review indicates the need for the following: (1) Investigations of other temperate and tropical symbiotic anemone species to assess the generality of trends seen in a few “model’ anemones. (2) Attention to the field ecology of temperate and tropical algal‐anemone symbioses, for example, how symbioses function under seasonally variable environmental factors and how zooxanthellae persist at high densities in darkness and winter. The greater stability of zooxanthella populations in temperate hosts may be useful to understanding tropical symbioses in which bleaching (loss of zooxanthellae) is of major concern. (3) Study of the evolutionary history of symbiosis in both temperate and tropical seas. Continued exploration of the phylogenetic relationships between host anemones and zooxanthella strains may show how and why zooxanthellae differ in anemone hosts in both environments.

The intertidal distribution of two algal symbionts hosted by Anthopleura xanthogrammica (Brandt 1835).

This paper quantifies the spatial distribution of zooxanthellae (ZX) and zoochlorellae (ZC), two algal symbionts common to the temperate anemone, Anthopleura xanthogrammica, in relation to shore height. Anemones in tidepools and crevices had varying algal proportions: >0.90 ZC (green anemones), 0.10 to 0.90 ZX (mixed), and >0.90 ZX (brown). Brown anemones are primarily found in the high intertidal and the upper region of tidepools. Mixed anemones are most common at intermediate shore heights and green anemones are exclusive to the low shore and at increasing depth in tidepools. Microhabitat was also important to algal proportion, as anemones in crevices had greater proportions of ZC than anemones in tidepools at the same shore height. In a reciprocal transplant experiment, A. xanthogrammica were moved between high and low shallow tidepools. All anemones moved from a low to a high tidepool exhibited a shift from ZC to ZX populations, while the anemones transplanted from high to low tidepools maintained ZX dominance. This is the first documentation that field algal populations can shift from ZC to ZX in Anthopleura. The field survey and transplant study results support the hypothesis that the relative abundance of ZX and ZC in A. xanthogrammica is influenced by the environmental gradient associated with shore height and microhabitat.

Carbonic anhydrase expression and synthesis in the sea anemone Anthopleura elegantissima are enhanced by the presence of dinoflagellate symbionts.

Endosymbiotic dinoflagellates resident within cnidarian hosts are extremely productive primary producers. This high productivity may be due in part to an inorganic carbon transport system, present in host tissue, that accelerates carbon delivery to the algae. The enzyme carbonic anhydrase (CA; EC 4.2.1.1) has been shown to be important in this transport system in a variety of tropical symbiotic cnidarians. This study extends the examination of CA to a temperate anemone, Anthopleura elegantissima, and documents symbiosis-enhanced production of CA at the biochemical and molecular level. Depending on light availability, A. elegantissima can occur naturally with (symbiotic) or without (aposymbiotic) dinoflagellates, making it an ideal study organism for symbiosis-enhanced gene expression. We compared (1) CA activities, (2) quantities of CA using an anti-human CA immunoprobe, and (3) quantities of transcript using a semiquantitative PCR in symbiotic versus aposymbiotic A. elegantissima host tissue. Amounts of activity, enzyme, and transcript were greatly enhanced in symbiotic animals compared with aposymbiotic animals. This is the first direct evidence that the presence of symbionts affects the expression of a host cnidarian gene. In addition, we report a full-length A. elegantissima CA cDNA sequence, obtained from subcloned reverse transcriptase-PCR products, and its relatedness to alpha-CAs from a variety of other metazoa, including higher vertebrates.

Mitotic indices of zooxanthellae: a comparison of techniques based on nuclear and cell division frequencies

Mitotic indices of zooxanthellae: a comparison of techniques based on nuclear and cell division frequencies