Class Anthozoa Research Articles

First description of algal mutualistic endosymbiosis in a black coral (Anthozoa: Antipatharia)

The class Anthozoa is the largest meta- zoan group forming mutualistic symbioses with mi- croalgae. These algal symbionts (most commonly dino - flagellates of the genus Symbiodinium) are distributed across most anthozoan orders. Records of algal cells in antipatharian (black coral) tissues have been reported, but no detailed descriptions of a mutualistic endosym- biosis exist. Here we report on zooxanthellate speci- mens of an unidentified black coral species within the genus Cirrhipathes that were collected from reef slopes at depths of 15 to 38 m in the Indonesian Archi- pelago. Symbionts were abundant (~10 7 symbionts cm -2 ) and ultrastructural analysis revealed the presence of a distinct symbiosome surrounding the algae, as well as algal reproduction inside the gastrodermal layer. Mol- ecular analysis revealed the algae to be closely related to the symbionts (Symbiodinium clade G) of clionid sponges. There was also evidence for additional sym- bionts in clade C at low abundance. Taken together, these findings (high abundance, taxonomic identity, presence of symbiosome, in situ reproduction, and depth distribution) strongly suggest that these algae are functioning as mutualists. This study confirms and describes the symbiosis between Symbiodinium and a black coral species of the genus Cirrhipathes, supports the pervasiveness of mutualisms among anthozoan taxa, and highlights the diversity and flexibility of these symbiotic associations in a poorly studied group.

Multi-colored homologs of the green fluorescent protein from hydromedusa Obelia sp.

The presence of green fluorescent protein (GFP) within the bioluminescent system of Obelia (Cnidaria, Hydrozoa, Campanulariidae) was inferred shortly after the discovery of GFP in Aequorea. Despite the enormous success of Aequorea GFP as a genetically encoded fluorescent label, Obelia GFP thus far has been defeating attempts to clone it from the hydroid life cycle stage. Here, we report cloning of three GFP-like fluorescent proteins (FPs) from Obelia medusa, representing cyan, green, and yellow spectral types. Such color diversity has never been detected outside class Anthozoa, suggesting a more general function for multi-colored fluorescence in cnidarians than has been previously hypothesized. An unusual property of the new FPs is the formation of large soluble complexes of well-defined sizes and molecular weights, corresponding to up to 128 individual polypeptides. This aligns well with the earlier observation that luminescence in Obelia, unlike in Aequorea, is localized within subcellular granules, which prompts further inquiry into the self-assembly properties of the new FPs and their interactions with the photoprotein. The discovery of Obelia FPs fills the four-decade-old gap in the knowledge of cnidarian bioluminescence and provides experimental material to further investigate the details of its molecular mechanism.

Antifouling activity by sea anemone (Heteractis magnifica and H. aurora) extracts against marine biofilm bacteria

Sea anemones (Actiniaria) are solitary, ocean-dwelling members of the phylum Cnidaria and the class Anthozoa. In this study, we screened antibacterial activity of two benthic sea anemones (Heteractis magnifica and H. aurora) collected from the Mandapam coast of southeast India. Crude extracts of the sea anemone were assayed against seven bacterial biofilms isolated from three different test panels. The crude ex- tract of H. magnifica showed a maximum inhibition zone of 18 mm against Pseudomonas sp. and Escherichia coli and a minimum inhibition zone of 3 mm against Pseudomonas aeruginosa, Micrococcus sp., and Bacillus cerens for methanol, acetone, and DCM extracts, respectively. The butanol extract of H. aurora showed a maximum inhibition zone of 23 mm against Vibrio parahaemolyticus, whereas the methanol extract revealed a minimum inhibition zone of 1 mm against V. parahaemolyticus. The present study revealed that the H. aurora extracts were more effective than those of H. magnifica and that the active compounds from the sea anemone can be used as antifouling compounds.

Complete mitochondrial genomes of two Japanese precious corals, Paracorallium japonicum and Corallium konojoi (Cnidaria, Octocorallia, Coralliidae): Notable differences in gene arrangement

Precious coral are taxonomically a group of corals that belong to the family Coralliidae within the order Alcyonacea, subclass Octocorallia, and class Anthozoa, whose skeletal axes are used for jewelry. They are distributed in the Mediterranean Sea and in waters adjacent to Japan, Taiwan, Midway Island and the Hawaiian Islands. The genus Corallium of the family Coralliidae was recently divided into two genera, Corallium and Paracorallium, based on morphological observations, but insufficient molecular evidence to support this classification has been presented to date. We determined for the first time the complete mitochondrial genome sequence of two precious corals P. japonicum and C. konojoi, in order to clarify their systematic positions. The circular mitochondrial genomes of P. japonicum and C. konojoi are 18,913 bp and 18,969 bp in length, respectively, and encode 13 typical energy pathway protein coding genes ( nad1– 6, nad4L, cox1– 3, cob, atp6 and atp8), two ribosomal RNA genes ( rns and rnl), a transfer RNA ( trnM) and a mismatch repair gene homologue msh1. The two genomes have an overall nucleotide sequence identity of 97.5%, which is comparable to that between Acanella eburnea and Keratoisidinae sp. belonging to Octocorallia. Surprisingly, however, their gene arrangements were not identical. Phylogenetic analyses using seven complete mitochondrial genome sequences belonging to species in the subclass Octocorallia indicated that within the subclass, at least three gene order rearrangement events occurred during evolution. Our results support the validity of the morphological classification that separated the family Coralliidae into two genera, Corallium and Paracorallium.

Decadal-scale changes in abundance of non-scleractinian invertebrates on a Caribbean coral reef

The benthic community structure of the shallow reefs of St. John, US Virgin Islands, was studied from 1992 to 2007 to test the hypotheses that the abundances of non-scleractinian invertebrates have changed, and further, that the changes are associated with variation in the percentage cover of scleractinian coral and macroalgae. The study utilized photoquadrats (0.25 m 2) from fringing reefs (7–9 m depth) characterized by igneous boulders and low (< 5%) coral cover, and results from six sites were pooled to describe these reefs as a single habitat. Photoquadrats were analyzed biennially for the abundance of invertebrates, particularly those belonging to four well represented classes (Anthozoa, Demospongiae, Echinoidea, and Polychaeta), as well as the cover of scleractinians and macroalgae. Overall, the combined (multivariate) abundance of 30 invertebrate taxa changed over time, the combined (multivariate) abundance of the four invertebrate classes changed over time, and the individual (univariate) abundance of anthozoans, sponges, and echinoids changed over time. Throughout the study, coral cover remained < 5%, and while it varied significantly, it did not display a consistent trajectory of change; in contrast, the cover of macroalgae increased throughout the study. While it is unsurprising that the abundances of invertebrates changed over 15 y, notably they varied even though coral cover remained stable, in only a few cases were they related positively to macroalgal cover, and in most cases it was members of the suspension feeding guild that became more abundant. These outcomes suggest that: (1) benthic invertebrates on the shallow reefs of St. John may be more strongly influenced by regional (e.g., larva supply) than local (e.g., coral cover) conditions, (2) Caribbean reefs have changed more since the early 1990s than can be inferred from variation in cover of coral and macroalgae, and (3) suspension feeding invertebrates have become more common on shallow fringing reefs in at least one location.

Remarks on the classification system of the Heliolitida

The concept of the sum of common features has been used in order to compare the Heliolitida with the Tabulata and Rugosa and determine the position of Heliolitida in the classification system of Palaeozoic corals. Such morphological characters as the number of septa, septa of the second order, epitheca, coenenchyme and multiplicity by four have been considered. Coenenchyme is present in many taxa and there are no more compelling reasons for the integration of Heliolitida and Halysitida than for the integration of Heliolitida with other taxa containing coenenchyme. The dynamics of some features shows that the differences between three main groups of Palaeozoic corals are within the limits of variation of relative organisms. A high degree of similarity testifies to the close relationship of the Heliolitida and the different qualitative and quantitative expression of similar features testifies to their early divergence from a common ancestor and subsequent independent evolution. Asto-phylogeny and morphological homogeny of all Heliolitida confirm their evolution as a separate, unitary group. Heliolitida is a monophyletic group of corals that is equal to the Tabulata and Rugosa in rank. A common origin requires that the orders should be united into one subclass Paleosclerocoralla of the class Anthozoa.

Evolution of Calcium-carbonate Skeletons in the Hydractiniidae

Biomineralization has mostly been studied in the class Anthozoa (Phylum Cnidaria), but very little is known about the evolution of the calcified skeleton in the class Hydrozoa or about the processes leading to its formation. The evolution of the calcified skeleton is here investigated in the hydrozoan family Hydractiniidae. A phylogenetic analysis of ribosomal, mitochondrial, and nuclear-protein-coding DNA sequences supported two independent origins of the calcified skeleton within the Hydractiniidae and indicates a case of parallel evolution, as suspected in the Anthozoa. Neither of the two origins of skeleton in the Hydractiniidae has led to either speciose or numerically abundant species, in contrast with other skeletonized hydrozoan families. Finally, we show that the origin of calcified skeletons in the Hydractiniidae is significantly correlated with the distribution of species with calcium carbonate granules within a polyp's gastrodermal cells. This suggests that the presence of these granules precedes the origin of a full skeleton.

Correlated evolution of sex and reproductive mode in corals (Anthozoa: Scleractinia).

Sexuality and reproductive mode are two fundamental life-history traits that exhibit largely unexplained macroevolutionary patterns among the major groups of multicellular organisms. For example, the cnidarian class Anthozoa (corals and anemones) is mainly comprised of gonochoric (separate sex) brooders or spawners, while one order, Scleractinia (skeleton-forming corals), appears to be mostly hermaphroditic spawners. Here, using the most complete phylogeny of scleractinians, we reconstruct how evolutionary transitions between sexual systems (gonochorism versus hermaphrodism) and reproductive modes (brooding versus spawning) have generated large-scale taxonomic patterns in these characters. Hermaphrodites have independently evolved in three large, distantly related lineages consisting of mostly reef-building species. Reproductive mode in corals has evolved at twice the rate of sexuality, while the evolution of sexuality has been heavily biased: gonochorism is over 100 times more likely to be lost than gained, and can only be acquired by brooders. This circuitous evolutionary pathway accounts for the prevalence of hermaphroditic spawners among reef-forming scleractinians, despite their ancient gonochoric heritage.

Coral immunology and resistance to disease

Scleractinian corals (phylum Cnidaria, class Anthozoa) have innate immunological responses against infections. Research has recently suggested that corals also possess an adaptive-like immunological repertoire that recognizes specific pathogens and allografts. While evolutionarily distinct, the corals' innate and adaptive-like immunity systems are not mutually exclusive because the phagocytic cells of the non-specific, innate immune system may activate specific adaptive immunological responses. Warming oceans may immunocompromise coral hosts, making them more susceptible to tropical marine diseases, independent of the virulence of the pathogen. The ability of corals to ward off both primary and opportunistic infections, through adaptive-like mechanisms, may play a critical role in the corals' ability to fight future disease infection. Here we show evidence that corals possess immunological repertoires that extend well beyond simple innate defenses. The extent to which corals have developed such an adaptive-like immune repertoire will determine whether corals will survive climate change and other anthropogenic disturbances.

Phylogenetic analysis of the Cnidaria

The recent members of the phylum Cnidaria were analyzed with phylogenetic methodology and the help of the PAUP Computer program. The Cnidaria are established as a monophylum by their cnidocysts, planula larva, and a polyp stage. The Ctenophora were seen as the most probable sister group of the Cnidaria. Arguments for the monophyly of the cnidarian classes Anthozoa, Scyphozoa, Cubozoa, and Hydrozoa were providea. For the ground plan of the Cnidaria the following characters were postulated: triphasic life cycle consisting of a planula larva, a benthic polyp stage, and a sexually propagating medusa like stage. For the polyp a radial symmetry, lack of septae, and hollow tentacles were assumed. The original medusa probably was tetraradial and developed from the polyp stage by a total metamorphosis. Twelve polarized characters were used to generate cladograms. The most parsimonious one showed the Anthozoa as the first offshoot of the tree with the united Scyphozoa, Cubozoa and Hydrozoa forming its sister group. Within this sister group the Scyphozoa and Cubozoa were seen as sistergroups to each other. Both groups united are then the sistergroup of the Hydrozoa. A bootstrap analysis yielded the same tree with high probabilities for the internal nodes. Despite assuming a planktonic origin of the Cnidaria in this investigation, the resulting cladogram is also compatible with an evolution of the medusa stage within the Cnidaria after the splitting-off of the Anthozoa. The possible loss of the medusa stage in the Anthozoa is discussed.

Sox genes in the coral Acropora millepora: divergent expression patterns reflect differences in developmental mechanisms within the Anthozoa

BackgroundSox genes encode transcription factors that function in a wide range of developmental processes across the animal kingdom. To better understand both the evolution of the Sox family and the roles of these genes in cnidarians, we are studying the Sox gene complement of the coral, Acropora millepora (Class Anthozoa).ResultsBased on overall domain structures and HMG box sequences, the Acropora Sox genes considered here clearly fall into four of the five major Sox classes. AmSoxC is expressed in the ectoderm during development, in cells whose morphology is consistent with their assignment as sensory neurons. The expression pattern of the Nematostella ortholog of this gene is broadly similar to that of AmSoxC, but there are subtle differences – for example, expression begins significantly earlier in Acropora than in Nematostella. During gastrulation, AmSoxBb and AmSoxB1 transcripts are detected only in the presumptive ectoderm while AmSoxE1 transcription is restricted to the presumptive endoderm, suggesting that these Sox genes might play roles in germ layer specification. A third type B Sox gene, AmSoxBa, and a Sox F gene AmSoxF also have complex and specific expression patterns during early development. Each of these genes has a clear Nematostella ortholog, but in several cases the expression pattern observed in Acropora differs significantly from that reported in Nematostella.ConclusionThese differences in expression patterns between Acropora and Nematostella largely reflect fundamental differences in developmental processes, underscoring the diversity of mechanisms within the anthozoan Sub-Class Hexacorallia (Zoantharia).

Diversity and evolution of coral fluorescent proteins.

GFP-like fluorescent proteins (FPs) are the key color determinants in reef-building corals (class Anthozoa, order Scleractinia) and are of considerable interest as potential genetically encoded fluorescent labels. Here we report 40 additional members of the GFP family from corals. There are three major paralogous lineages of coral FPs. One of them is retained in all sampled coral families and is responsible for the non-fluorescent purple-blue color, while each of the other two evolved a full complement of typical coral fluorescent colors (cyan, green, and red) and underwent sorting between coral groups. Among the newly cloned proteins are a “chromo-red” color type from Echinopora forskaliana (family Faviidae) and pink chromoprotein from Stylophora pistillata (Pocilloporidae), both evolving independently from the rest of coral chromoproteins. There are several cyan FPs that possess a novel kind of excitation spectrum indicating a neutral chromophore ground state, for which the residue E167 is responsible (numeration according to GFP from A. victoria). The chromoprotein from Acropora millepora is an unusual blue instead of purple, which is due to two mutations: S64C and S183T. We applied a novel probabilistic sampling approach to recreate the common ancestor of all coral FPs as well as the more derived common ancestor of three main fluorescent colors of the Faviina suborder. Both proteins were green such as found elsewhere outside class Anthozoa. Interestingly, a substantial fraction of the all-coral ancestral protein had a chromohore apparently locked in a non-fluorescent neutral state, which may reflect the transitional stage that enabled rapid color diversification early in the history of coral FPs. Our results highlight the extent of convergent or parallel evolution of the color diversity in corals, provide the foundation for experimental studies of evolutionary processes that led to color diversification, and enable a comparative analysis of structural determinants of different colors.

Genomic organization, gene structure, and developmental expression of three Clustered otx genes in the sea anemone Nematostella vectensis

Otx homeodomain transcription factors have been studied in a variety of eumetazoan animals where they have roles in anterior neural development, endomesoderm formation, and the formation of larval ciliated fields. Here, we describe the gene structure and developmental expression of three Otx loci in the starlet sea anemone, Nematostella vectensis (phylum Cnidaria; class Anthozoa). Nematostella's three Otx genes (OtxA, OtxB, and OtxC) are located in a compact genomic cluster spanning 63.6 kb. The homeodomains of all three Otx genes are highly similar to their bilaterian counterparts, but only OtxB exhibits the conserved WSP motif that is located downstream of the homeodomain in many Otx proteins. The genomic organization, in concert with phylogenetic analyses, indicates that two tandem duplications occurred in the lineage leading to Nematostella some time after the Cnidaria diverged from the Bilateria. In situ hybridization reveals that otx is initially expressed by invaginating mesendodermal cells in the gastrula. Later, each of the three otx paralogs is expressed in three discrete larval body regions: in the endoderm of the foot or physa, in an endodermal ring surrounding the pharynx, and in the ectoderm of the tentacles. These data suggest that a single otx locus had already acquired diverse developmental functions in the cnidarian-bilaterian ancestor. Furthermore, following two gene duplications in the line leading to Nematostella, there have been only minor alterations in the spatiotemporal expression of the three Otx paralogs. However, the absence of a conserved protein domain in OtxA and OtxC suggests functional evolution of the protein itself.

The innate immune repertoire in Cnidaria - ancestral complexity and stochastic gene loss

Analysis of genomic resources available for cnidarians revealed that several key components of the vertebrate innate immune repertoire are present in representatives of the basal cnidarian class Anthozoa, but are missing in Hydra, a member of the class Hydrozoa, indicating ancient origins for many components of the innate immune system.

Characterization of a novel EF-hand homologue, CnidEF, in the sea anemone Anthopleura elegantissima

The superfamily of EF-hand proteins is comprised of a large and diverse group of proteins that contain one or more characteristic EF-hand calcium-binding domains. This study describes and characterizes a novel EF-hand cDNA, CnidEF, from the sea anemone Anthopleura elegantissima (Phylum Cnidaria, Class Anthozoa). CnidEF was found to contain two EF-hand motifs near the C-terminus of the deduced amino acid sequence and two regions near the N-terminus that could represent degenerate EF-hand motifs. CnidEF homologues were also identified from two other sea anemone species. A combination of bioinformatic and molecular phylogenetic analyses was used to compare CnidEF to EF-hand proteins in other organisms. The closest homologues identified from these analyses were a luciferin binding protein (LBP) involved in the bioluminescence of the anthozoan Renilla reniformis, and a sarcoplasmic calcium-binding protein (SARC) involved in fluorescence of the annelid worm Nereis diversicolor. Predicted structure and folding analysis revealed a close association with bioluminescent aequorin (AEQ) proteins from the hydrozoan cnidarian Aequorea aequorea. Neighbor-joining analyses grouped CnidEF within the SARC lineage along with AEQ and other cnidarian bioluminescent proteins rather than in the lineage containing calmodulin (CAM) and troponin-C (TNC).

Components of both major axial patterning systems of the Bilateria are differentially expressed along the primary axis of a ‘radiate’ animal, the anthozoan cnidarian Acropora millepora

Cnidarians are animals with a single (oral/aboral) overt body axis and with origins that nominally predate bilaterality. To better understand the evolution of axial patterning mechanisms, we characterized genes from the coral, Acropora millepora (Class Anthozoa) that are considered to be unambiguous markers of the bilaterian anterior/posterior and dorsal/ventral axes. Homologs of Otx/otd and Emx/ems, definitive anterior markers across the Bilateria, are expressed at opposite ends of the Acropora larva; otxA-Am initially around the blastopore and later preferentially toward the oral end in the ectoderm, and emx-Am predominantly in putative neurons in the aboral half of the planula larva, in a domain overlapping that of cnox-2Am, a Gsh/ind gene. The Acropora homologs of Pax-3/7, NKX2.1/vnd and Msx/msh are expressed in axially restricted and largely non-overlapping patterns in larval ectoderm. In Acropora, components of both the D/V and A/P patterning systems of bilateral animals are therefore expressed in regionally restricted patterns along the single overt body axis of the planula larva, and two ‘anterior’ markers are expressed at opposite ends of the axis. Thus, although some specific gene functions appear to be conserved between cnidarians and higher animals, no simple relationship exists between axial patterning systems in the two groups.

A NEW GENUS OF EARLY CAMBRIAN CORAL IN ESMERALDA COUNTY, SOUTHWESTERN NEVADA

Numerous Early Cambrian corals or “coralomorphs,” as they are often classified, are recorded from North America, Australia, and Siberia. A new Early Cambrian coral, Harklessia yuenglingensis n. gen. and sp., is found in conjunction with archaeocyathan-microbial reefs in Esmeralda County, southwestern Nevada. The coral-bearing reefs are within quartzic, trilobite-rich packstone beds in the upper portion of the Harkless Formation (Bonnia–Olenellus Zone). Coralla are constructed by subpolygonal to polygonal, cerioid, close-packed corallite tubes. Coralla average 12 cm in height by 18 cm in diameter with individual corallite tubes ranging from 1.2 to 3.2 mm in diameter. Corallites are greater than 25 mm in length. Septa and tabulae are not present.Many of the Early Cambrian corals previously described have attributes of the class Anthozoa and subclass Zoantharia, with some specific similarities to tabulate corals. Harklessia yuenglingensis is placed confidently within the class Anthozoa, subclass Zoantharia because its morphological characteristics indicate an affinity to true corals, but whether H. yuenglingensis is a tabulate coral remains uncertain.

Fission in Sea Anemones: Integrative Studies of Life Cycle Evolution

Sea anemones (Phylum Cnidaria; Class Anthozoa, Order Actiniaria) exhibit a diversity of developmental patterns that include cloning by fission. Because natural histories of clonal and aclonal sea anemones are quite different, the gain and loss of fission is an important feature of actiniarian lineages. We have used mitochondrial DNA and nuclear intron DNA phylogenies to investigate the evolution of longitudinal fission in sixteen species in the genus Anthopleura, and reconstructed an aclonal ancestor that has given rise at least four times to clonal descendents. For A. elegantissima from the northeastern Pacific Ocean, a transition to clonality by fission was associated with an up-shore habitat shift, supporting prior hypotheses that clonal growth is an adaptation to the upper shore. Fission in Actiniaria likely precedes its advent in Anthopleura, and its repeated loss and gain is perplexing. Field studies of the acontiate sea anemone Aiptasia californica provided insight to the mechanisms that regulate fission: subtidal Aiptasia responded to experimentally destabilized substrata by increasing rates of pedal laceration. We put forth a general hypothesis for actiniarian fission in which sustained tissue stretch (a consequence of substratum instability or intrinsic behavior) induces tissue degradation, which in turn induces regeneration. The gain and loss of fission in Anthopleura lineages may only require the gain and loss of some form of stretching behavior. In this view, tissue stretch initiates a cascade of developmental events without requiring complex gene regulatory linkages.

Phospholipase A2 in Cnidaria

Phospholipase A2 (PLA2) is an enzyme present in snake and other venoms and body fluids. We measured PLA2 catalytic activity in tissue homogenates of 22 species representing the classes Anthozoa, Hydrozoa, Scyphozoa and Cubozoa of the phylum Cnidaria. High PLA2 levels were found in the hydrozoan fire coral Millepora sp. (median 735 U/g protein) and the stony coral Pocillopora damicornis (693 U/g) that cause skin irritation upon contact. High levels of PLA2 activity were also found in the acontia of the sea anemone Adamsia carciniopados (293 U/g). Acontia are long threads containing nematocysts and are used in defense and aggression by the animal. Tentacles of scyphozoan and cubozoan species had high PLA2 activity levels: those of the multitentacled box jellyfish Chironex fleckeri contained 184 U/g PLA2 activity. The functions of cnidarian PLA2 may include roles in the capture and digestion of prey and defense of the animal. The current observations support the idea that cnidarian PLA2 may participate in the sting site irritation and systemic envenomation syndrome resulting from contact with cnidarians.

Antifilarial activity of Zoanthus species (Phylum Coelenterata, Class Anthzoa) against human lymphatic filaria, Brugia malayi.

The chloroform methanol (1:1) extract of an unidentified green zoanthus (Phylum Coelenterata, Class Anthozoa) showed promising in vitro adulticidal activity with a lethal concentration of 125 microg/ml on Brugia malayi. This extract brought about a 52.2% reduction in circulating microfilariae of B. malayi when administered to infected Mastomys coucha at 250 mg/kg, orally for 5 consecutive days. Further fractionation of the extract led to the recovery of four fractions, which were evaluated simultaneously in both in vitro and in vivo systems against B. malayi. The chloroform fraction at 250 mg/kg orally for 5 days exhibited the highest macrofilaricidal action (42.5%), closely followed by the insoluble n-butanol fraction (34.3%), the soluble hexane fraction (32.4%), and the soluble n-butanol fraction (20.4%). In addition, the hexane soluble fraction caused 44.3% sterilization of the surviving female parasites. Two compounds isolated were found devoid of antifilarial activity.