The Genus Cerion (Gastropoda: Cerionidae) in the Florida Keys.

The systematic relationships and phylogeography of Cerion incanum, the only species of Cerion native to the Florida Keys, are reviewed based on partial sequences of the mitochondrial COI and 16S genes derived from 18 populations spanning the range of this species and including the type localities of all four described subspecies. Our samples included specimens of Cerion casablancae, a species introduced to Indian Key in 1912, and a population of C. incanum x C. casablancae hybrids descended from a population of C. casablancae introduced onto Bahia Honda Key in the same year. Molecular data did not support the partition of C. incanum into subspecies, nor could populations be apportioned reliably into subspecies based on morphological features used to define the subspecies. Phylogenetic analyses affirmed the derived relationship of C. incanum relative to other cerionids, and indicated a Bahamian origin for the Cerion fauna of southern Florida. Relationships among the populations throughout the Keys indicate that the northernmost populations, closest to the Tomeu paleoislands that had been inhabited by Cerion petuchi during the Calabrian Pleistocene, are the oldest. The range of Cerion incanum expanded as the archipelago that is the Florida Keys was formed since the lower Tarantian Pleistocene by extension from the northeast to the southwest, with new islands populated as they were formed. The faunas of the High Coral Keys in the northeast and the Oölite Keys in the southwest, both with large islands that host multiple discontinuous populations of Cerion, are each composed of well supported clades that are characterized by distinctive haplotypes. In contrast, the fauna of the intervening Low Coral Keys consist of a heterogeneous series of populations, some with haplotypes derived from the High Coral Keys, others from the Oölite Keys. Individuals from the C. incanum x C. casablancae hybrid population inhabiting the southeastern coast of Bahia Honda Key were readily segregated based on their mitogenome lineage, grouping either with C. incanum or with C. casablancae from Indian Key. Hybrids with C. casablancae mitogenomes had haplotypes that were more divergent from their parent mitogenome than were hybrids with C. incanum mitogenomes

Biodiversity estimates and ecological interpretations of meiofaunal communities are biased by the taxonomic approach

Accurate assessments of biodiversity are crucial to advising ecosystem-monitoring programs and understanding ecosystem function. Nevertheless, a standard operating procedure to assess biodiversity accurately and consistently has not been established. This is especially true for meiofauna, a diverse community (>20 phyla) of small benthic invertebrates that have fundamental ecological roles. Recent studies show that metabarcoding is a cost-effective and timeeffective method to estimate meiofauna biodiversity, in contrast to morphological-based taxonomy. Here, we compare biodiversity assessments of a diverse meiofaunal community derived by applying multiple taxonomic methods based on comparative morphology, molecular phylogenetic analysis, DNA barcoding of individual specimens, and metabarcoding of environmental DNA. We show that biodiversity estimates are strongly biased across taxonomic methods and phyla. Such biases affect understanding of community structures and ecological interpretations. This study supports the urgency of improving aspects of environmental high-throughput sequencing and the value of taxonomists in correctly understanding biodiversity estimates

Rapid Assessment of Octocoral Diversity and Habitat on Saba Bank, Netherlands Antilles

Saba Bank is a large submerged platform (∼2200 km2), average depth 30 m, located 4 km southwest of Saba Island in Netherlands Antilles, Caribbean Sea. Ships traveling to and from oil terminals on nearby St. Eustatius routinely anchor on the Bank, damaging benthic megafauna. Gorgonian octocorals are vulnerable to anchor damage, and they are common and conspicuous in shallow water (15–50 m) around the banks. This prompted a rapid assessment of octocoral habitat and diversity. The primary objectives were to estimate total species richness and to characterize habitats vis a vis gorgonians. Landsat imagery and multibeam bathymetry were employed to identify random sites for quantitative transects. A Seabotix LBV200L remotely operated vehicle (ROV) and SCUBA were used to collect and survey to 130 m. A total of 14 scuba dives and 3 ROV dives were completed in 10 days. During that time, 48 octocoral species were collected, including two likely undescribed species in the genera Pterogorgia and Lytreia. Gorgonian richness was exceptional, but not all species were collected, because the species accumulation curve remained steeply inclined after all surveys. Two shallow-water gorgonian habitat types were identified using multidimensional scaling and hierarchical cluster analyses: 1) a high diversity, high density fore-reef environment characterized by Eunicea spp., Gorgonia spp., and Pseudopterogorgia spp. and 2) a low diversity, low density plateau environment characterized by Pseudopterogorgia acerosa, Pterogorgia guadalupensis, and Gorgonia mariae. The analyses support hypotheses of broad (∼15 km) habitat homogeneity (ANOSIM, P>0.05), but a significant difference between fore-reef and plateau environments (ANOSIM, P<0.05). However, there was some indication of habitat heterogeneity along the 15 km study section of the 50 km platform edge along the southeast rim. Our results highlight the complexity and biodiversity of the Saba Bank, and emphasize the need for more scientific exploration

Phylogenetic Relationships in the Gorgonian Family Plexauridae (Anthozoa: Octocorallia: Holaxonia) Based on Two Mitochondrial Genes: Evidence for Multiple Lineages

Holaxonian octocorals are diverse and abundant on many marine hard substrates and, within this group, members of the Plexauridae are an important component of tropical reef assemblages. In the most recent morphological revision of octocorals, Bayer (1981) included the Paramuriceidae (as Stenogorgiinae) within the plexaurids based on a lack of distinguishing characters. As a result, the Plexauridae now comprises 31 genera and occurs throughout the tropics as well as at higher latitudes to depths of at least 900 m. To begin to understand historical relationships within this now large and diverse assemblage, and to test the monophyly of the family and some of its component genera, DNA sequences of two mitochondrial loci (msh1 and ND2, ~1212 bp) from 40 species in 21 genera from deep and shallow waters in the tropical western Atlantic, West and East Pacific (plus 5 taxa in the closely related Gorgoniidae and two outgroups) were analyzed. Results recover three strongly supported clades that correspond roughly to the Plexauridae, Paramuriceidae and Gorgoniidae, though their mutual relationships remain unclear. Representatives of several genera appear to be scattered among the 3 families ; e.g., Hypnogorgia sp. (Paramuriceidae) falls within a clade consisting of both Pacific and Atlantic Muricea spp. (Plexauridae), while Swiftia sp., Scleracis sp. and an Atlantic Thesea sp. (all Paramuriceidae) group with the gorgoniids. Some Atlantic and Pacific species of several plexaurid and paramuriceid genera were monophyletic (Muricea spp., Bebryce spp.), while others were not (Echinomuricea spp., Thesea spp., Villogorgia spp.). These molecular results indicate that current octocoral taxonomy needs revision

Molecular Ecology and Evolution of Reef Corals and their Algal Symbionts

Molecular approaches to the fields of taxonomy, systematics and population genetics are imperative in order to better understand the evolutionary history and ecological interactions among organisms. This dissertation utilized genetic techniques to address evolutionary and ecological questions in corals and octocorals at different levels of resolution, from ancient phylogenetic relationships to the relatively recent population dynamics of individuals within species in both the coral host and its algal endosymbionts (genus Symbiodinium). Collectively, this work: (1) demonstrates a novel use of genes involved in the calcification process to test hypotheses of morphological convergence in scleractinian corals; (2) helps reconcile contrasting morphological and molecular phylogenetic perspectives among closely related species of a gorgonian octocoral; and (3) elucidates the genetic connectivity, dispersal potential, host acquisition, and specificity between a reef coral and its algal symbionts. Results show that the phylogenetic history of two genes involved in calcification in scleractinian corals is complex and consists of species-specific gene duplications and losses. However, in at least some cases, these calcification gene phylogenies agreed with taxonomic hypotheses based on morphology, in contrast to standard molecular phylogenies of neutral genes not involved in calcification. Second, molecular data support calyx morphology as a valid systematic character among species within the gorgonian octocoral genus Pterogorgia, but do not corroborate branch morphology as a useful systematic character. In addition, hybridization and/or morphological plasticity likely play important roles in shaping the morphological species described within this genus. Finally, populations of the gorgonian octocoral Plexaura flexuosa and its algal symbiont, Symbiodinium B1, showed different genetic structure across the Florida reef tract and Caribbean. P. flexuosa displayed high genetic connectivity over distances up to ~1900 km, while Symbiodinium B1 showed clear population subdivisions among sites separated by \u3c100 km, as well as within sites (\u3c100 m). Despite this structure, local environmental pools of Symbiodinium B1 are genetically diverse, and there is evidence for limited dispersal of Symbiodinium vectored by the larval stage of their coral hosts over ecological timescales. Together, data from the host (P. flexuosa) and its symbiont suggest high resilience potential, mediated by a passive process of stochastic host acquisition and dispersal. Additionally, in contrast to the very high specificity previously reported for gorgonian octocorals and Symbiodinium, genotypic clones (and genetically similar genotypes) of P. flexuosa did not show specificity to particular genotypes of Symbiodinium B1, and vice versa. Future work should continue to further integrate molecular techniques to establish a more comprehensive understanding of the ecology and evolution of these important organisms and the ecosystems they build

Molecular evolution of calcification genes in morphologically similar but phylogenetically unrelated scleractinian corals

Molecular phylogenies of scleractinian corals often fail to agree with traditional phylogenies derived from morphological characters. These discrepancies are generally attributed to non-homologous or morphologically plastic characters used in taxonomic descriptions. Consequently, morphological convergence of coral skeletons among phylogenetically unrelated groups is considered to be the major evolutionary process confounding molecular and morphological hypotheses. A strategy that may help identify cases of convergence and/or diversification in coral morphology is to compare phylogenies of existing "neutral" genetic markers used to estimate genealogic phylogenetic history with phylogenies generated from non-neutral genes involved in calcification (biomineralization). We tested the hypothesis that differences among calcification gene phylogenies with respect to the "neutral" trees may represent convergent or divergent functional strategies among calcification gene proteins that may correlate to aspects of coral skeletal morphology. Partial sequences of two nuclear genes previously determined to be involved in the calcification process in corals, "Cnidaria-III" membrane-bound/secreted α-carbonic anhydrase (CIII-MBSα-CA) and bone morphogenic protein (BMP) 2/4, were PCR-amplified, cloned and sequenced from 31 scleractinian coral species in 26 genera and 9 families. For comparison, "neutral" gene phylogenies were generated from sequences from two protein-coding "non-calcification" genes, one nuclear (β-tubulin) and one mitochondrial (cytochrome b), from the same individuals. Cloned CIII-MBSα-CA sequences were found to be non-neutral, and phylogenetic analyses revealed CIII-MBSα-CAs to exhibit a complex evolutionary history with clones distributed between at least 2 putative gene copies. However, for several coral taxa only one gene copy was recovered. With CIII-MBSα-CA, several recovered clades grouped taxa that differed from the "non-calcification" loci. In some cases, these taxa shared aspects of their skeletal morphology (i.e., convergence or diversification relative to the "non-calcification" loci), but in other cases they did not. For example, the "non-calcification" loci recovered Atlantic and Pacific mussids as separate evolutionary lineages, whereas with CIII-MBSα-CA, clones of two species of Atlantic mussids (Isophyllia sinuosa and Mycetophyllia sp.) and two species of Pacific mussids (Acanthastrea echinata and Lobophyllia hemprichii) were united in a distinct clade (except for one individual of Mycetophyllia). However, this clade also contained other taxa which were not unambiguously correlated with morphological features. BMP2/4 also contained clones that likely represent different gene copies. However, many of the sequences showed no significant deviation from neutrality, and reconstructed phylogenies were similar to the "non-calcification" tree topologies with a few exceptions. Although individual calcification genes are unlikely to precisely explain the diverse morphological features exhibited by scleractinian corals, this study demonstrates an approach for identifying cases where morphological taxonomy may have been misled by convergent and/or divergent molecular evolutionary processes in corals. Studies such as this may help illuminate our understanding of the likely complex evolution of genes involved in the calcification process, and enhance our knowledge of the natural history and biodiversity within this central ecological group

A phylogenetic analysis of the Primnoidae (Anthozoa: Octocorallia: Calcaxonia) with analyses of character evolution and a key to the genera and subgenera

Abstract Background Previous phylogenetic analyses of primnoid octocorals utilizing morphological or molecular data have each recovered evolutionary relationships among genera that are largely incongruent with each other, with some exceptions. In an effort to reconcile molecular-based phylogenies with morphological characters, phylogenetic reconstructions were performed with 33 of 43 primnoid genera using four loci (mtMutS, COI, 28S and 18S), and ancestral state reconstructions were performed using 9 taxonomically relevant characters. In addition, an updated illustrated key to the current 48 genus-level (43 genera, 5 subgenera) primnoids is presented. Results Ancestral state reconstruction recovered the ancestral colony shape of primnoids as dichotomous planar. Convergence was detected among all 9 characters, and reversals to the character state of the common ancestor occurred in 4 characters. However, some characters were found to be informative. For example, the weak ascus scale of Metafannyella is not likely homologous to the ascus scales of Onogorgia and Fannyella, and the monophyly of two subgenera within Thouarella, which contain polyps in either whorls or an isolated arrangement, was supported. Phylogenetic analyses were generally consistent with previous studies, and resulted in the synonymy of one genus and a subgenus, the elevation of two subgenera, and the transfer of two species back to an original genus. For example, body wall ornamentation of Fanellia was re-evaluated, indicating a synonymy with Callogorgia; the utility of polyp arrangement for the subgenus Plumarella (Dicholaphis) was not supported, and is synonymized with the nominate subgenus Plumarella (Plumarella); the subgenera Plumarella (Faxiella) and Plumarella (Verticillata) are raised to generic status; and the two Plumarella species (P. diadema and P. undulata) are transferred back to Thouarella based on the homology of their marginal scales. Conclusions Altogether, and similar to other octocorallian groups, these results indicate that many of the morphological characters examined among primnoids, particularly colony morphology, are labile and exhibit complex evolutionary histories. However, some morphological characters such as coordination of polyps, presence of the ascus body wall scale, number of rows of body wall scales, and number of marginal scales help identify many clades, and are suitable for robust systematic assessments among primnoids

Molecular and Morphological Species Boundaries in the Gorgonian Octocoral Genus Pterogorgia (Octocorallia: Gorgoniidae).

Most gorgonian octocoral species are described using diagnostic characteristics of their sclerites (microscopic skeletal components). Species in the genus Pterogorgia, however, are separated primarily by differences in their calyx and branch morphology. Specimens of a morphologically unusual Pterogorgia collected from Saba Bank in the NE Caribbean Sea were found with calyx morphology similar to P. citrina and branch morphology similar to P. guadalupensis. In order to test morphological species boundaries, and the validity of calyx and branch morphology as systematic characters, a phylogenetic analysis was undertaken utilizing partial gene fragments of three mitochondrial (mtMutS, cytochrome b, and igr4; 726bp total) and two nuclear (ITS2, 166bp; and SRP54 intron, 143bp) loci. The datasets for nuclear and mitochondrial loci contained few phylogenetically informative sites, and tree topologies did not resolve any of the morphological species as monophyletic groups. Instead, the mitochondrial loci and SRP54 each recovered two clades but were slightly incongruent, with a few individuals of P. guadalupensis represented in both clades with SRP54. A concatenated dataset of these loci grouped all P. anceps and P. guadalupensis in a clade, and P. citrina and the Pterogorgia sp. from Saba Bank in a sister clade, but with minimal variation/resolution within each clade. However, in common with other octocoral taxa, the limited genetic variation may not have been able to resolve whether branch variation represents intraspecific variation or separate species. Therefore, these results suggest that there are at least two phylogenetic lineages of Pterogorgia at the species level, and the atypical Pterogorgia sp. may represent an unusual morphotype of P. citrina, possibly endemic to Saba Bank. Branch morphology does not appear to be a reliable morphological character to differentiate Pterogorgia species (e.g., branches "flat" or "3-4 edges" in P. guadalupensis and P. anceps, respectively), and a re-evaluation of species-level characters (e.g., sclerites) is needed

Cladogram depicting hypothesized phylogenetic relationships among <i>Pterogorgia</i> species using morphological characters.

<p><i>P</i>. <i>anceps</i> and <i>P</i>. <i>guadalupensis</i> share the synapomorphic character “calyces within a common groove”. Autapomorphic characters, branches “3–4 edges in cross section” define <i>P</i>. <i>anceps</i> while branches “flat in cross section (7mm+ in width)” separate <i>P</i>. <i>guadalupensis</i>. <i>P</i>. <i>citrina</i> and the morphotype from Saba Bank share the synapomorphic character “distinct calyces”. <i>P</i>. <i>citrina</i> contains the autapomorphic character of branches “flat to slightly oval in cross section”, while the Saba Bank morphotype contains branches “flat in cross section (7mm+ in width)”. Drawings adapted from [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133517#pone.0133517.ref007" target="_blank">7</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133517#pone.0133517.ref023" target="_blank">23</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133517#pone.0133517.ref057" target="_blank">57</a>].</p