Historical navigation routes in European waters leave their footprint on the contemporary seascape genetics of a colonial urochordate.

Humans have intensively sailed the Mediterranean and European Atlantic waters throughout history, from the upper Paleolithic until today and centuries of human seafaring have established complex coastal and cross-seas navigation networks. Historical literature revealed three major long-lasting maritime routes (eastern, western, northern) with four commencing locations (Alexandria, Venice, Genoa, Gibraltar) and a fourth route (circum-Italian) that connected between them. Due to oceangoing and technological constraints, most voyages were coastal, lasted weeks to months, with extended resting periods, allowing the development of fouling organisms on ship hulls. One of the abiding travellers in maritime routes is the colonial ascidian Botryllus schlosseri already known since the eighteenth century in European and Mediterranean ports. This species, was almost certainly one of the common hull fouling travellers in all trade routes for centuries. Employing COI haplotypes (1008 samples) and microsatellite alleles (995 samples) on colonies sampled from 64 pan-European sites, present-day Botryllus populations in the Mediterranean Sea/European Atlantic revealed significant segregation between all four maritime routes with a conspicuous partition of the northern route. These results reveal that past anthropogenic transports of sedentary marine species throughout millennia long seafaring have left their footprint on contemporary seascape genetics of marine organisms.

Ambiguities in the taxonomic assignment and species delineation of botryllid ascidians from the Israeli Mediterranean and other coastlines.

Based on mtCOI sequences comparisons, recent studies reassigned the 'dwarf Botrylloides leachii' from the Levant as Botrylloides nigrum. Here we conducted a survey of the literature and of deposited mtCOI sequences of botryllid ascidians, elucidating ambiguities in their taxonomy. We found that the species, dwarf morph of Botrylloides leachii, Botrylloides nigrum, Botryllus aster and Botryllus arenaceus are grouped together on a single molecular taxon. Then, results of three additional markers (18S, 28S, H3) contradicted literature suggestions, revealing minute distances between Botrylloides leachii and the 'dwarf Botrylloides leachii'. Moreover, only Botrylloides leachii and the 'dwarf Botrylloides leachii' develop giant ampullae as an allorecognition response. Our results raise the possibility that inadequate identification, together with faults in molecular assignment, including queries regarding the efficacy of the mtCOI as the exclusive barcoding tool in botryllid ascidians, is the major culprits responsible for the emerged inconsistencies between the mtCOI sequences and traditional taxonomy. Thus, we assign the Levantine dwarf form as Botrylloides aff. leachii.

Phylogenetics, biogeography and population genetics of the ascidian Botryllus schlosseri in the Mediterranean Sea and beyond.

The wide distribution of the ascidian Botryllus schlosseri along the Mediterranean coasts has been documented since the eighteenth century. However, despite copious documentation, analyses of dispersal modes and genetic profiles were limited to local populations or restricted regions. In order to get a pan-Mediterranean overview, 288 specimens from 11 populations of B. schlosseri from the western and eastern Mediterranean basins were sampled and analyzed using five microsatellite loci and COI sequences. Both molecular markers revealed high polymorphisms, with 182 microsatellites alleles and 54 COI haplotypes. Overall, Fst, Dest, and COI Фpt values were 0.146, 0.635 and 0.322, respectively, reflecting a high genetic diversity and a significant genetic structure as compared to other B. schlosseri populations worldwide, reflected by substantially higher values for effective number of alleles (Ne) in the Mediterranean. A phylogenetic analysis of the COI sequences resulted in four distinct clades and two molecular operational taxonomic units (OTUs). We recorded a stronger genetic structure among the populations of the eastern basin compared to the western basin (microsatellites Fst=0.217 versus 0.082; COI Фpt=0.416 versus 0.171), suggesting either a restricted connectivity between the basins or a stronger genetic drift in each basin. The occurrence of two OTUs and different ecological conditions may also contribute to this finding. Mean Nei's genetic distance in the eastern Mediterranean populations was more than three times higher compared to the western basin. No correlation was observed between geographic and genetic distances (Mantel test), suggesting that maritime transport is the main dispersal vector of B. schlosseri colonies. The possibility that the Mediterranean is a center of diversity for B. schlosseri, and probably its site of origin, is further discussed.