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Injury response is key to successful regeneration. Yet, transcriptome analyses of injury response were performed only on a handful of regenerative organisms. Here, we studied the injury response of the solitary ascidian Polycarpa mytiligera, an emerging model system, capable of regenerating any body part. We used the siphon as a model for studying transcriptional changes following injury, and identified genes that were activated in the initial 24 hours post amputation (hpa).Highly conserved genes, such as bone morphogenetic protein-1 (BMP1), growth hormone secretagogue receptor (GHSR) and IL-17, were upregulated by 12 hpa, yet their expression was sustained only in non-regenerating tissue fragments. We optimized fluorescent in situ hybridization, and found that the majority of BMP1+ cells were localized to the rigid tunic that covers the animal. This highlights the importance of this tissue, particularly during injury response. BMP1 was overexpressed following injuries to other body regions, suggesting that it was a part of a common injury-induced program.Our study suggests that, initially, specific injury-induced genes were upregulated in P. mytiligera organs, yet, later, a unique transcriptional profile was observed only in regenerating tissues. These findings highlight the importance of studying diverse regenerating and non-regenerating organisms for complete understanding of regeneration.

Understanding how neurons regenerate following injury remains a central challenge in regenerative medicine. Adult mammals have a very limited ability to regenerate new neurons in the central nervous system (CNS). In contrast, the basal chordate Polycarpa mytiligera can regenerate its entire CNS within seven days of complete removal. Transcriptome sequencing, cellular labeling, and proliferation in vivo essays revealed that CNS regeneration is mediated by a newly formed neural progeny and the activation of neurodevelopmental pathways that are associated with enhanced stem-cell activity. Analyzing the expression of 239 activated pathways enabled a quantitative understanding of gene-set enrichment patterns at key regeneration stages. The molecular and cellular mechanisms controlling the regenerative ability that this study reveals can be used to develop innovative approaches to enhancing neurogenesis in closely-related chordate species, including humans.

Plastic additives (PAs) are chemical compounds incorporated into the plastic during the manufacturing process. Phthalate acid esters, bisphenols, and nonylphenols are all PAs found in marine environments and associated with endocrine-disrupting processes. However, our knowledge regarding the impact of endocrine-disrupting PAs on coral-reef organisms is limited. As reef population structure is directly linked to reproduction and larval settlement processes, interference with hormonal systems can impact coral-reef community structure, particularly if the effects of PAs differ among species. In the current study we exposed the reproductive products of four tropical coral-reef invertebrates to environmentally-relevant concentrations of four prevalent PAs in seawater: dibutyl phthalate (DBP), dimethyl phthalate, (DMP), 4-nonylphenol (4-NP), and bisphenol A (BPA), as well as to 10

The shipping industry is considered the main vector of introduction of marine non-indigenous species (NIS). NIS distributions are often a consequence of frequent trade activities that are affected by economic trends. A dominant trend in the shipping industry is the operation of Ultra Large Container Vessels (ULCV), which are over 395 m long and sail mostly on the East-Asia - northern-Europe route. Understanding the risk of NIS introduction by this emerging shipping category is needed for devising strategies for sustainable shipping. Here, we conducted a controlled simulation of key abiotic factors that determine marine bioinvasion success: temperature, salinity, and food availability along selected routes, under two treatments: ULCV and intermediate-size vessels. We tested the effect of each treatment and the varying environmental conditions on the survival of two invasive ascidians (Chordata, Ascidiacea). We used survival analysis methods to locate predictors of ascidian mortality; Environmental conditions at ports with high mortality were used to identify similar major ports on a global scale as potential abiotic barriers. The key factors in ascidian mortality varied between the two species, but for both species, the treatment and salinity were dominant predictors for survival. We identified Port Klang, Rotterdam, and Dammam as ports with high mortality and located several globally distributed major ports that present similar environmental conditions. Our results highlight the potential role of selected major ports as abiotic barriers to fouling organisms during ocean voyages. The tolerance of the tropical-origin Microcosmus exasperatus to the northern-Europe conditions, and of the temperate/sub-tropical origin Styela plicata, to high temperature conditions, point out the urgent need to modify international fouling regulations in view of global change. Further studies on the survival of fouling organisms during a cascade of changing environmental conditions will contribute to the advancement of science-based regulations to reduce the adverse effects of NIS.

The increasing use of pharmaceuticals in human and veterinary medicine, along with their poor removal rates at wastewater treatment facilities is resulting in the chronic release of pharmaceutically-active compounds (PhACs) into the marine environment, where they pose a threat to non-target organisms. A useful approach, as applied in the current study for assessing the effects of PhACs on non-target organisms, is the proteomic approach: the large-scale study of an organism's proteins. Using ‘shotgun’ proteomics, we identified differentially-expressed proteins based on peptide fragments in the solitary ascidian, Herdmania momus, following a 14-day laboratory experimental exposure to the PhAC carbamazepine (CBZ), an anticonvulsant and antidepressant medication, frequently detected in the aquatic environment. Individuals were exposed to environmentally relevant concentrations: 5 or 10 µg/L of CBZ, in addition to a control treatment. Out of 199 identified proteins, 24 were differentially expressed (12%) between the treatment groups, and thus can potentially be developed as biomarkers for CBZ contamination. Ascidians’ phylogenetic position within the closest sister group to vertebrates presents an advantage in examining the pathological effects of PhACs on vertebrate-related organs and systems. Together with the world-wide distribution of some model ascidian species, and their ability to flourish in pristine and polluted sites, they provide a promising tool through which to study the extent and effects of PhAC contamination on marine organisms.

Significance The mechanisms by which organisms form mineralized skeletons have been a major research focus for the last 50 y and remain so today. Among the most surprising discoveries is the recent observation that different animals use the same mechanisms, and precisely the same amorphous precursors, to form biomineralized structures as diverse as coral skeletons, molluscan shells, and sea urchin spines. In living animals, skeletal biomineralization from amorphous precursors correlates with a distinctive nanoparticulate texture that can be preserved in fossils, enabling us to probe mechanisms of skeletal formation in early animals. We document nanoparticulate texture in some of the oldest known carbonate skeletons, which strongly suggests that skeletons formed from amorphous precursors throughout the recorded history of animals., Crystallization by particle attachment (CPA) of amorphous precursors has been demonstrated in modern biomineralized skeletons across a broad phylogenetic range of animals. Precisely the same precursors, hydrated (ACC-H2O) and anhydrous calcium carbonate (ACC), have been observed spectromicroscopically in echinoderms, mollusks, and cnidarians, phyla drawn from the 3 major clades of eumetazoans. Scanning electron microscopy (SEM) here also shows evidence of CPA in tunicate chordates. This is surprising, as species in these clades have no common ancestor that formed a mineralized skeleton and appear to have evolved carbonate biomineralization independently millions of years after their late Neoproterozoic divergence. Here we correlate the occurrence of CPA from ACC precursor particles with nanoparticulate fabric and then use the latter to investigate the antiquity of the former. SEM images of early biominerals from Ediacaran and Cambrian shelly fossils show that these early calcifiers used attachment of ACC particles to form their biominerals. The convergent evolution of biomineral CPA may have been dictated by the same thermodynamics and kinetics as we observe today.

Global increase in the use of pharmaceutically-active compounds (PhACs), and their insufficient removal in wastewater treatment plants, have resulted in their continuous release into the marine environment. We investigated the use of the solitary ascidians Herdmania momus, Microcosmus exasperatus, and Styela plicata as bioindicators of three common PhACs in the Israeli coastal waters: Bezafibrate, carbamazepine and diclofenac. Both the Mediterranean and the Red-Sea coasts were found contaminated with PhACs, detected at all 11 sampling sites, with four sites contaminated with all three studied PhACs. Diclofenac was most frequent, present in nine of the 11 sites with concentrations reaching 51.9 ng/g of dry weight sample (dw). Bezafibrate and carbamazepine reached concentrations of 47.8 ng/g dw and 14.3 ng/g dw, respectively. The alarming detection of such high concentrations of PhACs in ascidians along Israel's coasts demonstrates both the extent of PhACs contamination in the region, and the potential of ascidians as bioindicators, and emphasizes the urgent need for additional research into PhAC contamination sources and effects.

Background Adaptive evolution is one of the crucial mechanisms for organisms to survive and thrive in new environments. Recent studies suggest that adaptive evolution could rapidly occur in species to respond to novel environments or environmental challenges during range expansion. However, for environmental adaptation, many studies successfully detected phenotypic features associated with local environments, but did not provide ample genetic evidence on microevolutionary dynamics. It is therefore crucial to thoroughly investigate the genetic basis of rapid microevolution in response to environmental changes, in particular on what genes and associated variation are responsible for environmental challenges. Here, we genotyped genome-wide gene-associated microsatellites to detect genetic signatures of rapid microevolution of a marine tunicate invader, Ciona robusta, during recent range expansion to the harsh environment in the Red Sea. Results The Red Sea population was significantly differentiated from the other global populations. The genome-wide scan, as well as multiple analytical methods, successfully identified a set of adaptive genes. Interestingly, the allele frequency largely varied at several adaptive loci in the Red Sea population, and we found significant correlations between allele frequency and local environmental factors at these adaptive loci. Furthermore, a set of genes were annotated to get involved in local temperature and salinity adaptation, and the identified adaptive genes may largely contribute to the invasion success to harsh environments. Conclusions All the evidence obtained in this study clearly showed that environment-driven selection had left detectable signatures in the genome of Ciona robusta within a few generations. Such a rapid microevolutionary process is largely responsible for the harsh environmental adaptation and therefore contributes to invasion success in different aquatic ecosystems with largely varied environmental factors. Electronic supplementary material The online version of this article (10.1186/s12862-018-1311-1) contains supplementary material, which is available to authorized users.

The interaction of microplastic (MP) with marine organisms is crucial for understanding the significant effect that MP and its additives may have on marine environments. However, knowledge regarding the magnitude of these pollutants in the Eastern Mediterranean Sea and the tropical Red Sea is still scarce. Here we examined the levels of phthalate acid esters (PAEs) and MP in Herdmania momus and Microcosmus exasperatus sampled along the Mediterranean and Red Sea coasts of Israel. High levels of dibutyl phthalate (DBP) and bis (2‑ethylhexyl) phthalate (DEHP) were found in ascidians at the majority of sampling sites, and MP particles were found in ascidians at all sites. As efficient filter-feeders and being widely-distributed, invasive ascidians present fundamental opportunities for the environmental monitoring of microplastic and its additives worldwide. The high levels of pollutants revealed emphasize the need for further research into the magnitude and effects of MP and PAEs in these regions.

Regeneration, the process of replacing lost or damaged body parts, has long captured human imagination and is a key feature among all animal phyla. Due to their close phylogenetic relationship to vertebrates and their high regenerative abilities, ascidians (Chordata, Ascidiacea) are often used as models to shed light on the cellular and genetic process involved in tissue regeneration. Surprisingly, ascidian regeneration studies are based on only a few model species. In this chapter, we point out the important potential of solitary ascidians in regenerative and stem cell studies. We review recent studies of regeneration among solitary ascidians and discuss the cellular mechanism of tissue regeneration and the possible involvement of circulating cells in these processes. New data regarding the relationship between age and regeneration abilities of the solitary ascidian Polycarpa mytiligera (Stolidobranchia, Styelidae) are presented. The unique regeneration abilities found in P. mytiligera following evisceration of its digestive system and following amputation of its neural complex and siphon-associated structures and nerves imply on its potential to serve as a novel model system for understanding tissue regeneration.

Ascidians (Chordata, Ascidiacea) are considered to be prominent marine invaders, able to tolerate highly polluted environments and fluctuations in salinity and temperature. Here, we examined the seasonal and spatial dynamics of the microbial communities in the inner-tunic of two invasive ascidians, Styela plicata (Lesueur 1823) and Herdmania momus (Savigny 1816), in order to investigate the changes that occur in the microbiome of non-indigenous ascidians in different environments. Microbial communities were characterized using next-generation sequencing of partial (V4) 16S rRNA gene sequences. A clear differentiation between the ascidian-associated microbiome and bacterioplankton was observed, and two distinct sets of operational taxonomic units (OTUs), one core and the other dynamic, were recovered from both species. The relative abundance of the dynamic OTUs in H. momus was higher than in S. plicata, for which core OTU structure was maintained independently of location. Ten and seventeen core OTUs were identified in S. plicata and H. momus, respectively, including taxa with reported capabilities of carbon fixing, ammonia oxidization, denitrification, and heavy-metal processing. The ascidian-sourced dynamic OTUs clustered in response to site and season but significantly differed from the bacterioplankton community structure. These findings suggest that the associations between invasive ascidians and their symbionts may enhance host functionality while maintaining host adaptability to changing environmental conditions.

Harbor systems represent passive gateways for the introduction of nonnative ascidians that compete with the surrounding benthos and may spread through localized dispersal, even populating adjacent natural reefs. To investigate the potential role of microbial symbionts in the success of ascidian introductions and spread, we evaluated the host-specificity of prokaryotic communities within two ascidian species commonly found off the North Carolina coast. Replicate samples of the native ascidian Eudistoma capsulatum, the nonnative ascidian Distaplia bermudensis and seawater were collected from artificial (harbor) and natural reef substrates. Prokaryotic communities in seawater samples and ascidian tunics were characterized via next-generation sequencing of partial 16S rRNA gene sequences. Ascidian microbiomes clustered strongly in response to host species, with significant differences in community structure between the two species and seawater. Further, symbiont community structure differed significantly between E. capsulatumindividuals collected from artificial and natural habitats, though this was not the case for D. bermudensis. These findings suggested that some ascidian species possess stable microbial symbiont communities that allow them to thrive in a wide range of habitats, while other species rely on the restructuring of their microbial communities with specific symbionts (e.g. Chelativorans) to survive under particular environmental conditions such as increased pollution.

Many ascidian species have experienced worldwide introductions, exhibiting remarkable success in crossing geographic borders and adapting to local environmental conditions. To investigate the potential role of microbial symbionts in these introductions, we examined the microbial communities of three ascidian species common in North Carolina harbors. Replicate samples of the globally introduced species Distaplia bermudensis, Polyandrocarpa anguinea, and P. zorritensis (n = 5), and ambient seawater (n = 4), were collected in Wrightsville Beach, NC. Microbial communities were characterized by next-generation (Illumina) sequencing of partial (V4) 16S rRNA gene sequences. Ascidians hosted diverse symbiont communities, consisting of 5,696 unique microbial OTUs (at 97% sequenced identity) from 44 bacterial and three archaeal phyla. Permutational multivariate analyses of variance revealed clear differentiation of ascidian symbionts compared to seawater bacterioplankton, and distinct microbial communities inhabiting each ascidian species. 103 universal core OTUs (present in all ascidian replicates) were identified, including taxa previously described in marine invertebrate microbiomes with possible links to ammonia-oxidization, denitrification, pathogenesis, and heavy-metal processing. These results suggest ascidian microbial symbionts exhibit a high degree of host-specificity, forming intimate associations that may contribute to host adaptation to new environments via expanded tolerance thresholds and enhanced holobiont function.

Invasive ascidians (Chordata, Tunicata) are dominant nuisance organisms. The current study investigated the role of marine vessels in their dispersal and introduction. An examination of 45 dry-docked marine vessels, comprising recreational, commercial, and military craft, in five Israeli shipyards along the Mediterranean coast, revealed non-indigenous ascidians (NIA) on every second vessel investigated. Military vessels featured the highest ascidian abundance and richness, potentially related to their maintenance routine. Niche areas on the vessels such as sea chests and the propeller exhibited the highest occurrence of ascidians. Overall, these findings provide strong evidence that marine vessels play an acute role in NIA introduction and dispersal, with military vessels and niche areas on all the vessels being more susceptible to serving as vectors. A discovery of a new introduced species during the surveys suggests that the monitoring of marine vessels can serve as an effective tool for the early detection of NIA.

Ascidians or sea squirts form a diverse group within chordates, which includes a few thousand members of marine sessile filter-feeding animals. Their mitochondrial genomes are characterized by particularly high evolutionary rates and rampant gene rearrangements. This extreme variability complicates standard polymerase chain reaction (PCR) based techniques for molecular characterization studies, and consequently only a few complete Ascidian mitochondrial genome sequences are available. Using the standard PCR and Sanger sequencing approach, we produced the mitochondrial genome of Ascidiella aspersa only after a great effort. In contrast, we produced five additional mitogenomes (Botrylloides aff. leachii, Halocynthia spinosa, Polycarpa mytiligera, Pyura gangelion, and Rhodosoma turcicum) with a novel strategy, consisting in sequencing the pooled total DNA samples of these five species using one Illumina HiSeq 2000 flow cell lane. Each mitogenome was efficiently assembled in a single contig using de novo transcriptome assembly, as de novo genome assembly generally performed poorly for this task. Each of the new six mitogenomes presents a different and novel gene order, showing that no syntenic block has been conserved at the ordinal level (in Stolidobranchia and in Phlebobranchia). Phylogenetic analyses support the paraphyly of both Ascidiacea and Phlebobranchia, with Thaliacea nested inside Phlebobranchia, although the deepest nodes of the Phlebobranchia-Thaliacea clade are not well resolved. The strategy described here thus provides a cost-effective approach to obtain complete mitogenomes characterized by a highly plastic gene order and a fast nucleotide/amino acid substitution rate.

Phylum Hemichordata, composed of worm-like Enteropneusta and colonial Pterobranchia, has been reported to only contain about 100 species. However, recent studies of hemichordate phylogeny and taxonomy suggest the species number has been largely underestimated. One issue is that species must be described by experts, and historically few taxonomists have studied this group of marine invertebrates. Despite this previous lack of coverage, interest in hemichordates has piqued in the past couple of decades, as they are critical to understanding the evolution of chordates–as acorn worms likely resemble the deuterostome ancestor more closely than any other extant animal. This review provides an overview of our current knowledge of hemichordates, focusing specifically on their global biodiversity, geographic distribution, and taxonomy. Using information available in the World Register of Marine Species and published literature, we assembled a list of 130 described, extant species. The majority (83%) of these species are enteropneusts, and more taxonomic descriptions are forthcoming. Ptychoderidae contained the greatest number of species (41 species), closely followed by Harrimaniidae (40 species), of the recognized hemichordate families. Hemichordates are found throughout the world’s oceans, with the highest reported numbers by regions with marine labs and diligent taxonomic efforts (e.g. North Pacific and North Atlantic). Pterobranchs are abundant in Antarctica, but have also been found at lower latitudes. We consider this a baseline report and expect new species of Hemichordata will continue to be discovered and described as new marine habitats are characterized and explored.

Appeltans, Ward ... et al., Background: The question of how many marine species exist is important because it provides a metric for how much we do and do not know about life in the oceans. We have compiled the first register of the marine species of the world and used this baseline to estimate how many more species, partitioned among all major eukaryotic groups, may be discovered. Results: There are ∿226,000 eukaryotic marine species described. More species were described in the past decade (∿20,000) than in any previous one. The number of authors describing new species has been increasing at a faster rate than the number of new species described in the past six decades. We report that there are ∿170,000 synonyms, that 58,000-72,000 species are collected but not yet described, and that 482,000-741,000 more species have yet to be sampled. Molecular methods may add tens of thousands of cryptic species. Thus, there may be 0.7-1.0 million marine species. Past rates of description of new species indicate there may be 0.5 ± 0.2 million marine species. On average 37% (median 31%) of species in over 100 recent field studies around the world might be new to science. Conclusions: Currently, between one-third and two-thirds of marine species may be undescribed, and previous estimates of there being well over one million marine species appear highly unlikely. More species than ever before are being described annually by an increasing number of authors. If the current trend continues, most species will be discovered this century. © 2012 Elsevier Ltd., WoRMS has benefited from funding as part of several EU projects: Network of Excellence in Marine Biodiversity and Ecosystem Functioning (MarBEF), Pan-European Species directories Infrastructure (PESI), Distributed Dynamic Diversity Databases for Life (4D4Life), the Global Biodiversity Information Facility (GBIF), and the Census of Marine Life (CoML). It originated in the European Register of Marine Species (ERMS) that was funded by the EU Marine Science and Technology (MAST) research program

Symbiotic relationships between shrimps and other invertebrates are a very common phenomenon in tropical environments. Although the caridean shrimp-ascidian association has been known for many years, the nature of this relationship is still unclear. The current study investigated the association between the caridean shrimp Odontonia sibogae (Bruce, 1972) and solitary ascidians. A combination of field work conducted along the Red Sea coast of Israel and laboratory experiments, conducted during 2015-2016, revealed a clear preference of the shrimps for the ascidian species Herdmania momus (Savigny, 1816), with a low survival ability of the shrimp outside their host's body. The shrimps usually inhabit their host as pairs of male and female or pair of females, but never as pairs of males. Out of the 53 studied females, 51% were observed to bear between 156-1,146 embryos, throughout the course of the year. As these ascidian hosts are known to create large aggregates, we suggest that males may possibly wander among the ascidians occupied by females in order to increase their reproductive success. To date, this is the first study to record the shrimp Dactylonia ascidicola (Borradaile, 1898) inhabiting the ascidian H. momus; and the first study to investigate in depth the ascidian-shrimp association in the Red Sea. It thus provides a platform for future research into the physiological and behavioral adaptations required for such a unique association.

Although ascidians are conspicuous members of the fouling community not much is known regarding their recruitment patterns in coral reefs. A 1-year study was carried out along the Red Sea coast of Israel to examine the effects of season and spatial distribution on ascidian recruitment to artificial marine structures. In general, autumn and spring were characterized by higher coverage with a significantly higher percentage of cover of Didemnum granulatum in autumn and higher numbers of Herdmania momus in spring. These species contributed the most to similarity between treatments consequently setting the pattern for each group (colonial and solitary). Halocynthia spinosa had significantly higher numbers during winter and Phallusia nigra was absent in spring and winter. H. momus showed a preference for horizontal surfaces. P. nigra and Ascidia cannelata showed a preference for floating units. It is concluded that the ascidian recruitment patterns are species-specific and vary between seasons, orientation and position on the substrata and in the water column.

The order Aplousobranchia (Chordata, Ascidiacea) contains approximately 1500 species distributed worldwide. Their phylogeny, however, remains unclear, with unresolved family relationships. While most Aplousobranchia are colonial, debates exist concerning the phylogenetic position of families such as the Diazonidae and Cionidae, which exhibit a solitary lifestyle and share morphological characteristics with both Aplousobranchia and Phlebobranchia orders. To clarify the phylogenetic position of the Diazonidae and Cionidae, we determined the complete mitochondrial sequence of the solitary diazonid Rhopalaea idoneta. The phylogenetic reconstruction based on the 13 mitochondrial protein coding genes strongly supports a positioning of Diazonidae well-nested within the Aplousobranchia rather than a positioning as a sister clade of the Aplousobranchia. In addition, we examined the regenerative ability of R. idoneta. Similar to colonial Aplousobranchia, R. idoneta was found to be able to completely regenerate its thorax. Ciona, also known to possess high regenerative abilities, is the Aplousobranchia sister clade rather than a member of the Phlebobranchia. Our results thus indicate that the colonial lifestyle was acquired in the Aplousobranchia, starting from a Ciona-like solitary ancestor and secondarily lost in Diazonidae representatives such as Rhopalaea. The solitary lifestyle of Rhopalaea is thus a derived characteristic rather than an ancestral trait.

The ejection of internal organs, i.e., evisceration, is a well-known phenomenon in sea-cucumbers. We report the ability of a member of the Chordate phyla, the tropical ascidian Polycarpa mytiligera, to eviscerate and regenerate its gut within 12 days and to rebuild its branchial sac within 19 days. Evisceration occurred within 4–43 seconds of gentle mechanical pressure exerted on the tunic in 47% of the tested P. mytiligera. Individuals were able to discard up to 3/4 of their digestive tract via the incurrent siphon by rupture of the branchial sac in this area. Although chemical analysis revealed no significant levels of toxic compounds, the eviscerated guts were unpalatable to the triggerfish and pufferfish on which they were tested, suggesting evisceration as a defense mechanism. Given the close affinity of ascidians to vertebrates, the regeneration pathway of the viscera and branchial sac of ascidians suggests its potential beneficial application in soft tissue regeneration research.

During regular surveys and collection of ascidians along the Red Sea coast of Israel, several specimens of an undescribed species of Rhopalaea were collected. Samples were collected by SCUBA from the natural coral reef and man-made structures at depths between 10 to 40m. This is the first species of the genus Rhopalaea described from the Red Sea, which is characterized by a transparent, uncolored gelatinous tunic with elongated attachment extensions, and is distinguished by its eight atrial lobes, thoracic muscle arrangement, and branchial sac structure.

9 páginas, 4 tablas, 4 figuras., Although marine biological invasions via the Suez Canal have been extensively documented, little is known about the introduction of non-indigenous ascidians (Chordata, Ascidiacea), a group containing particularly aggressive invasive species. Here, we used a multidisciplinary approach to study the introduction of the ascidianHerdmania momus into the Mediterranean Sea. We reviewed its taxonomy and global distribution, and analyzed how genetic variation is partitioned between sides of the Suez Canal. The taxonomic revision showed that H. momus currently has a wide Indo-Pacific distribution. Genetic data indicated two well-differentiated colonization histories across the eastern Mediterranean. Our findings suggest that the range expansion of H. momus has been greatly facilitated by the combined effect of human-mediated transport and the species’ ability to adapt to different environments. The integrative approach presented here is critical to attain a holistic understanding of marine biological invasions, especially when studying groups with a poorly resolved taxonomy., The research was funded by the Dan-David Foundation, the Steinhardt National Collections of Natural History (Zoological Museum, Tel Aviv University) the Israeli Taxonomy Initiative fellowship to N.S., and the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement no. PIOF-GA-2009-254634 to M.R.

16 pages, 5 figures, 1 table, 3 additional files., [Background] Tunicates have been recently revealed to be the closest living relatives of vertebrates. Yet, with more than 2500 described species, details of their evolutionary history are still obscure. From a molecular point of view, tunicate phylogenetic relationships have been mostly studied based on analyses of 18S rRNA sequences, which indicate several major clades at odds with the traditional class-level arrangements. Nonetheless, substantial uncertainty remains about the phylogenetic relationships and taxonomic status of key groups such as the Aplousobranchia, Appendicularia, and Thaliacea., [Results] Thirty new complete 18S rRNA sequences were acquired from previously unsampled tunicate species, with special focus on groups presenting high evolutionary rate. The updated 18S rRNA dataset has been aligned with respect to the constraint on homology imposed by the rRNA secondary structure. A probabilistic framework of phylogenetic reconstruction was adopted to accommodate the particular evolutionary dynamics of this ribosomal marker. Detailed Bayesian analyses were conducted under the non-parametric CAT mixture model accounting for site-specific heterogeneity of the evolutionary process, and under RNA-specific doublet models accommodating the occurrence of compensatory substitutions in stem regions. Our results support the division of tunicates into three major clades: 1) Phlebobranchia + Thaliacea + Aplousobranchia, 2) Appendicularia, and 3) Stolidobranchia, but the position of Appendicularia could not be firmly resolved. Our study additionally reveals that most Aplousobranchia evolve at extremely high rates involving changes in secondary structure of their 18S rRNA, with the exception of the family Clavelinidae, which appears to be slowly evolving. This extreme rate heterogeneity precluded resolving with certainty the exact phylogenetic placement of Aplousobranchia. Finally, the best fitting secondary-structure and CAT-mixture models suggest a sister-group relationship between Salpida and Pyrosomatida within Thaliacea., [Conclusion] An updated phylogenetic framework for tunicates is provided based on phylogenetic analyses using the most realistic evolutionary models currently available for ribosomal molecules and an unprecedented taxonomic sampling. Detailed analyses of the 18S rRNA gene allowed a clear definition of the major tunicate groups and revealed contrasting evolutionary dynamics among major lineages. The resolving power of this gene nevertheless appears limited within the clades composed of Phlebobranchia + Thaliacea + Aplousobranchia and Pyuridae + Styelidae, which were delineated as spots of low resolution. These limitations underline the need to develop new nuclear markers in order to further resolve the phylogeny of this keystone group in chordate evolution., This study is part of a Research Networks Program in Bioinformatics funded by the High Council for Scientific and Technological Cooperation between France and Israel (Grant # 3-3449 to DH, EJPD and YL). The present work was carried out as part of the Galathea 3 expedition under the auspices of the Danish Expedition Foundation (Dansk Ekspeditionsfond). This is Galathea 3 contribution no. P42 and also contributes to the Census of Marine Zooplankton. This work benefited from the bioinformatics cluster of the Institut des Sciences de l'Evolution de Montpellier. This is contribution ISEM 2009-88 of the Institut des Sciences de l'Evolution de Montpellier (UMR 5554 – CNRS).

Background Tunicates represent a key metazoan group as the sister-group of vertebrates within chordates. The six complete mitochondrial genomes available so far for tunicates have revealed distinctive features. Extensive gene rearrangements and particularly high evolutionary rates have been evidenced with regard to other chordates. This peculiar evolutionary dynamics has hampered the reconstruction of tunicate phylogenetic relationships within chordates based on mitogenomic data. Results In order to further understand the atypical evolutionary dynamics of the mitochondrial genome of tunicates, we determined the complete sequence of the solitary ascidian Herdmania momus. This genome from a stolidobranch ascidian presents the typical tunicate gene content with 13 protein-coding genes, 2 rRNAs and 24 tRNAs which are all encoded on the same strand. However, it also presents a novel gene arrangement, highlighting the extreme plasticity of gene order observed in tunicate mitochondrial genomes. Probabilistic phylogenetic inferences were conducted on the concatenation of the 13 mitochondrial protein-coding genes from representatives of major metazoan phyla. We show that whereas standard homogeneous amino acid models support an artefactual sister position of tunicates relative to all other bilaterians, the CAT and CAT+BP site- and time-heterogeneous mixture models place tunicates as the sister-group of vertebrates within monophyletic chordates. Moreover, the reference phylogeny indicates that tunicate mitochondrial genomes have experienced a drastic acceleration in their evolutionary rate that equally affects protein-coding and ribosomal-RNA genes. Conclusion This is the first mitogenomic study supporting the new chordate phylogeny revealed by recent phylogenomic analyses. It illustrates the beneficial effects of an increased taxon sampling coupled with the use of more realistic amino acid substitution models for the reconstruction of animal phylogeny.

The class Ascidiacea presents fundamental opportunities for research in the fields of development, evolution, ecology, natural products and more. This review provides a comprehensive overview of the current knowledge regarding the global biodiversity of the class Ascidiacea, focusing in their taxonomy, main regions of biodiversity, and distribution patterns. Based on analysis of the literature and the species registered in the online World Register of Marine Species, we assembled a list of 2815 described species. The highest number of species and families is found in the order Aplousobranchia. Didemnidae and Styelidae families have the highest number of species with more than 500 within each group. Sixty percent of described species are colonial. Species richness is highest in tropical regions, where colonial species predominate. In higher latitudes solitary species gradually contribute more to the total species richness. We emphasize the strong association between species richness and sampling efforts, and discuss the risks of invasive species. Our inventory is certainly incomplete as the ascidian fauna in many areas around the world is relatively poorly known, and many new species continue to be discovered and described each year.