Mitochondrial DNA lineages in the European populations of mussels (<Emphasis Type="Italic">Mytilus</Emphasis> spp.)

Marine mussels (Mytilus spp.) belong to a group of benthic species crucial to coastal ecosystems in Europe and are important for the cultivation industry. In the present study, the nuclear adhesive protein marker (Me15/16) was used for identification of Mytilus species in coastal areas, on a large geographic scale in Europe. Pure M. edulis populations were found in the White Sea and Iceland. M. edulis, M. trossulus and their hybrids were found in the Baltic Sea and the North Sea (Oosterschelde, The Netherlands). M. galloprovincialis, M. edulis and their hybrids occurred in Ireland. M. galloprovincialis populations were observed in the Sea of Azov (Black Sea), the Mediterranean and Portugal. The mitochondrial (mt) DNA coding-region ND2-COIII was studied by PCR (polymerase chain reaction) and RFLP (restriction fragment length polymorphism) assay methods. The mtDNA control region was studied by PCR. Substantial differentiation in the frequency of female haplotypes among the studied populations in Europe was observed. Despite isolation between the Mytilus taxa on a macro-geographic scale, considerable mitochondrial gene flow occurred between populations, with introgression in hybrid zones on a more local geographic scale in Europe. MtDNA of the Atlantic Iberian (Portugal) population of M. galloprovincialis was more similar to mtDNA in populations of M. galloprovincialis and M. edulis from the Atlantic coasts of the Ireland and M. edulis from the North Sea, than to M. galloprovincialis from the Mediterranean. Lower polymorphism of mtDNA in populations of the Baltic and Azov Sea mussels in comparison with other European populations was observed and can be explained by the recent history of both seas after the Pleistocene glaciation. In the M. galloprovincialis population from the Azov Sea, the presence of the male-inherited (M) genome was demonstrated for the first time by sequencing the control region and was observed at high frequency. Possible influence of mussel culture on geographic distribution of the Mytilus taxa in Europe is discussed.     

Phylogeography of the calanoid copepods <Emphasis Type="Italic">Calanus helgolandicus</Emphasis> and <Emphasis Type="Italic">C. euxinus</Emphasis> suggests Pleistocene divergences between Atlantic,Mediterranean, and Black Sea populations

Molecular systematic analyses of marine taxa are crucial for recording ocean biodiversity, so too are elucidation of the history of population divergence and the dynamics of speciation. In this paper we present the joined phylogeography of the calanoid copepod Calanus helgolandicus (Claus 1863) from the North East (NE) Atlantic and the Adriatic Sea and the closely related C. euxinus (Hulsemann 1991) from the Black Sea based on sequences of a mitochondrial Cytochrome Oxidase subunit I (COI) fragment. Coalescent-based Bayesian methods and minimum spanning networks are used to reconstruct the history of population divergence. Our results reveal that copepod populations from all three basins share a great number of haplotypes and demonstrate a close genetic affinity of C. euxinus with C. helgolandicus. The data do not support significant genetic structuring among samples within seas. Coalescent analyses suggest divergences between NE Atlantic, Mediterranean, and Black Sea populations dating back to the middle Pleistocene, with the NE Atlantic–Mediterranean divergence being the earliest and the Mediterranean–Black Sea divergence the most recent. These middle Pleistocene dates are much older than the estimated dates of colonisation of the Mediterranean and Black Seas based on paleoclimatic scenarios. Our results do not rule out that the assumed colonisations took place but they indicate that the populations colonising the Mediterranean and the Black Sea were already, and have since remained, diverged. The chaetognath Sagitta setosa, which has a comparable distribution pattern and feeds upon the copepods, provides a unique opportunity to compare phylogeographic patterns and distinguish among alternative hypotheses. The dates produced in this paper are in agreement with those estimated elsewhere for S. setosa. We propose that a great deal of the genetic make-up of marine planktonic populations comprises divergences that date back to long before the last glacial maximum. We consider questions on the taxonomic status of C. euxinus to remain open. However, its high genetic affinity to the C. helgolandicus calls for further investigation.     

Mytilus trossulus in Northern Europe

From data on allozyme, nuclear DNA and mitochondrial DNA markers, we show that the originally North Pacific/Northwest Atlantic mussel Mytilus trossulus is widespread on North European coasts, earliM. trossuluser thought to be inhabited only by Mytilus edulis. Several local occurrences of , interspersed with a dominant M. edulis, were recorded on the North Sea, Norwegian Sea and Barents Sea coasts of Norway and the Barents and White Sea coasts of Kola Peninsula in Russia. The proportion of M. trossulus genetic background observed at any one site varied from 0 to 95%. These new occurrences are not related to the previously known, introgressed M. trossulus population that occupies the Baltic Sea. The new northern occurrences retain both the F and M M. trossulus mitochondria, which have been lost from the Baltic stock. While hybridization takes place wherever M. trossulus and M. edulis meet, the extent of hybrization varies between the different contact areas. Hybrids are rare, and the hybrid zones are bimodal in the northern areas; more interbreeding has taken place further south in Norway, but even there genotypic disequilibria are higher than those in the steep transition zone between the Baltic mussel and M. edulis: there is no evidence of a collapse toward a hybrid swarm unlike in the Baltic. The Barents and White Sea M. trossulus are genetically slightly closer to the NW Atlantic than NE Pacific populations, while the Baltic mussel has unique features distinguishing it from the others. We postulate that the presence of M. trossulus in Northern Europe is a result of repeated independent inter- or transoceanic cryptic invasions of various ages, up to recent times.     

Colonization of the northwest Atlantic by the blue mussel, <Emphasis Type="Italic">Mytilus trossulus</Emphasis> postdates the last glacial maximum

Blue mussels in the genus Mytilus first arrived in the Atlantic Ocean from the Pacific during the Pliocene, following the opening of the Bering Strait. Repeated periods of glaciation throughout the Pleistocene led to re-isolation of the two ocean basins and the allopatric divergence of Mytilus edulis in the Atlantic and M. trossulus in the Pacific. Mytilus trossulus has subsequently colonized the northwest Atlantic (NW Atlantic) so that the two species are presently sympatric and hybridize throughout much of the Canadian Maritimes and the Gulf of Maine. To estimate when M. trossulus arrived in the NW Atlantic, we have examined sequence variation within a portion of the female mtDNA lineage large untranslated region (F-LUR) for 156 mussels sampled from three Pacific and eleven Atlantic populations of M. trossulus. Although we found no evidence of reciprocal monophyly for Pacific and NW Atlantic M. trossulus, limited gene flow between ocean basins has led to the divergence of unique sequence clades within each ocean basin. In contrast, relative genetic homogeneity indicates high levels of gene flow within each basin. Coalescence-based analysis of the F-LUR sequences suggests that M. trossulus recolonized the NW Atlantic from the northeast Pacific subsequent to a demographic expansion in the Pacific that occurred ~96,000 years before present (ybp). Estimates of timing of divergence for Pacific and NW Atlantic populations and the time since expansion among NW Atlantic sequence clades indicate that M. trossulus arrived in the NW Atlantic more recently, between 20,000 and 46,000 ybp. Given that these estimates overlap with the dates of peak ice in the NW Atlantic during the last glacial maximum (LGM, ~18,000–21,000 ybp), we suggest that colonization of the NW Atlantic by M. trossulus occurred during, but more likely just subsequent to, the LGM and was followed by rapid temporal and spatial expansion in the region.     

Inter-oceanic genetic differentiation among albacore (<Emphasis Type="Italic">Thunnus alalunga</Emphasis>) populations

Albacore (Thunnus alalunga) is a highly migratory pelagic species distributed in all tropical and temperate oceans. Recent analyses using both mitochondrial DNA (mtDNA) and nuclear DNA markers have demonstrated genetic subdivision within and between Atlantic and Pacific populations. However, although numerous biological differences have been reported for Atlantic and Mediterranean albacore, the genetic differentiation for these basins has not been demonstrated. We characterized 373 base pairs of nucleotide sequence from the mitochondrial DNA control region of 134 individuals collected in the Pacific (n=30), the northeast (NE) Atlantic (n=54) and the Mediterranean (n=50). Analysis of molecular variance (AMOVA) revealed a small, but highly significant, proportion of genetic variation separating these three regions (_st =0.041; P=0.009), a pattern also supported by pairwise comparisons. These results demonstrate for the first time the genetic distinctiveness of the Mediterranean albacore from the NE Atlantic population giving support to the current management practices based on separate units. This outcome is concordant with reported migratory movements related to reproductive behavior between the NE Atlantic and the Mediterranean Sea. Additionally, the phylogenetic analyses of DNA sequences revealed the presence of a shallow genetic discontinuity with no geographic association. These two phylogroups are more likely the result of the demographic history of this species (i.e. long demographic stable history) as opposed to historical vicariance as has been proposed for other highly migratory fishes.Communicated by P.W. Sammarco, Chauvin     

Phylogeography of surfclams,<Emphasis Type="Italic"> Spisula solidissima</Emphasis>, in the western North Atlantic based on mitochondrial and nuclear DNA sequences

The Atlantic surfclam, Spisula solidissima (Dillwyn), is broadly distributed in sandy sediments of the western North Atlantic between the Gulf of St. Lawrence and the Gulf of Mexico. In the United States, a substantial commercial fishery between Long Island and Cape Hatteras harvests offshore populations of one subspecies, S. s. solidissima. A smaller coastal form, S. s. similis Say (also known as S. s. raveneli Conrad), has a partially sympatric geographic distribution, but differs in several life-history characteristics. DNA sequence variation in mitochondrial cytochrome oxidase I (COI) and in introns at two nuclear calmodulin loci was examined to measure genetic divergence between the two subspecies and to test for population structure among populations of S. s. solidissima. Surfclams were collected from seven localities between 1994 and 2001. Based on both mitochondrial and nuclear DNA variation, the two subspecies of S. solidissima are reciprocally monophyletic, with a net COI divergence of 13.9%, indicating long-term reproductive isolation. The only significant differentiation among populations of S. s. solidissima (based on an AMOVA analysis of COI sequences) was between the Gulf of St. Lawrence and more southerly populations. A long internal branch in the S. s. solidissima genealogy coupled with low haplotype diversity in the northern-most population suggests that populations north and south of Nova Scotia have been isolated from each other in the past, with gene exchange more recently. Populations of S. s. similis from Atlantic and Gulf of Mexico coasts had a net COI divergence of 9.2%. Thus, diversification of Spisula spp. clams in the western North Atlantic involved an early adaptive divergence between coastal and offshore forms, with later barriers to dispersal emerging in the offshore form from north to south and in the coastal form between Atlantic and Gulf of Mexico populations.     

Active pelagic migrations of the bivalve Macoma balthica are dangerous

The bivalve Macoma balthica migrates twice during the benthic part of its life cycle. During the spring migration (May-June), the newly settled spat (0-group) migrates to the nurseries in the high intertidal. Seven to nine months later, the bivalves migrate back to the low tidal flats and the subtidal (winter migration, 1-group). Both 0- and 1-group M. balthica use byssus threads for active pelagic migrations. As many M. balthica disappear during these migrations, we examined experimentally the importance of predation on 0- and 1-group M. balthica. Laboratory experiments using a circular aquarium determined predation rates on buried (no current) and drifting (current) 0- and 1-group M. balthica by several fish species (plaice, flounder, goby and whiting) and the shore crab. Under illuminated conditions, more M. balthica were consumed when migrating than when buried, whereas there was no difference between experiments in conditions of darkness. For the 0-group, predation rates on migrating and buried M. balthica in the dark were lower than in the light. The stomachs of pelagic fish in the Wadden Sea and Oosterschelde estuary did not contain M. balthica during winter migration. In the Wadden Sea, 1-group M. balthica primarily migrated at night. In conclusion, enhanced predation on drifting, as compared to buried, M. balthica may be the mechanism that explains enhanced mortality during migration in light, and may explain why M. balthica mainly migrates at night in the field. As we found no M. balthica in stomachs of pelagic fish, we do not know whether predation on byssus drifting M. balthica exists in the field. There are, however, some indications for fish predation on infaunal polychaetes during pelagic migrations.     

Genetic structure of the flounders Platichthys flesus and P. stellatus at different geographic scales

The genetic structure of the flounders Platichthys flesus L. and P. stellatus Pallas was investigated on different spatial scales through analysis of allozyme variation at 7 to 24 polymorphic loci in samples collected from different regions (Baltic Sea, North Sea, Brittany, Portugal, western Mediterranean, Adriatic Sea, Aegean Sea and Japan) in 1984 to 1987. No geographic variation was evident within a region. Some pattern of differentiation by distance was inferred within the Atlantic, while the Mediterranean comprised three geographically isolated populations and was itself geographically isolated from the Atlantic (fixed allele differences at up to three loci were found among P. flesus populations from the Atlantic, the western Mediterranean, the Adriatic Sea, the Aegean Sea and also P. stellatus from the coast of Japan). Sea temperature during the reproductive period probably acts as a barrier to gene flow between populations. Genetic distances among European flounder populations (P. flesus) were higher than, or of the same magnitude as, the genetic distance between Pacific (P. stellatus) and European flounder populations, suggesting that P. flesus is paraphyletic and/or there is no phylogenetic basis to recognising P. stellatus as a different species. The divergence between P. flesus and P. stellatus was thus inferred to be more recent than the divergence between the present P. flesus populations from the NE Atlantic and eastern Mediterranean. The eastern Mediterranean populations are thought to originate from the colonisation of the Mediterranean by a proto-P. flesus/P. stellatus ancestor, whereas the present western Mediterranean population has undergone a more recent colonisation event by P. flesus. Patterns of mitochondrial DNA variation, established on a smaller array of P. flesus samples, were in accordance with the geographic patterns inferred from the allozyme survey. In addition, they supported the hypothesis of a two-step colonisation of the western Mediterranean. These results contribute to our understanding of the biogeography of the Mediterranean marine fauna, especially the group of boreal remnants to which P. flesus belongs. Received: 7 February 1997 / Accepted: 26 March 1997     

Genetic heterogeneity among <Emphasis Type="Italic">Eurytemora affinis</Emphasis> populations in Western Europe

Evolutionary diversification of the broadly distributed copepod sibling species complex Eurytemora affinis has been documented in the northern hemisphere. However, the fine scale geographic distribution, levels of genetic subdivision, evolutionary, and demographic histories of European populations have been less explored. To gain information on genetic subdivision and to evaluate heterogeneity among European populations, we analyzed samples from 8 locations from 58° to 45°N and 0° to 23°E, using 549 base pairs of the mitochondrial cytochrome oxidase subunit I (COI) gene. We discovered three distinct lineages of E. affinis in Western Europe, namely the East Atlantic lineage, the North Sea/English Channel (NSEC) lineage, and the Baltic lineage. These geographically separated lineages showed sequences divergence of 1.7–2.1%, dating back 1.9 million years (CI: 0.9–3.0 My) with no indication of isolation by distance. Genetic divergence in Europe was much lower than among North American lineages. Interestingly, genetic structure varied distinctively among the three lineages: the East Atlantic lineage was divided between the Gironde and the Loire populations, the NSEC lineage comprised one single population unit spanning the Seine, Scheldt and Elbe rivers and the third lineage was restricted to the Baltic Proper (Sweden). We revealed high haplotype diversity in the East Atlantic and the Baltic lineages, whereas in the NSEC lineage haplotype diversity was comparatively low. All three lineages showed signs of at least one demographic expansion event during Pleistocene glaciations that marked their genetic structure. These results provide a preliminary overview of the genetic structure of E. affinis in Europe.     

Genetic affinities of the bivalve Macoma balthica from the Pacific coast of North America: evidence for recent introduction and historical distribution

The tellinid bivalve Macoma balthica (L.) has an extensive geographic range that reaches from temperate to arctic coastal waters in the North Atlantic and North Pacific oceans. Recent studies have indicated that eastern and western North Atlantic populations are morphologically and genetically different from one another, and that they may have diverged as sibling species. To determine the genetic relationship between M. balthica from the Pacific and Atlantic coasts of North America, populations from each coast were examined at 11 enzyme loci using standard starch gel electrophoresis. Allele frequency data indicate that M. balthica populations from San Francisco Bay, California appear more closely related to western North Atlantic populations than to populations from Oregon. We suggest that San Francisco Bay populations were introduced relatively recently from western North Atlantic populations. The Oregon populations are probably a natural extension of northern populations that occur along Northern Asia and in the eastern North Atlantic.     

Discrimination among and variation within species of Pseudocalanus based on the GPI locus

Patterns of allozyme variation at the GPI (glucose phosphate isomerase) locus showed that the recently described or clarified Pseudocalanus newmani, P. acuspes, P. moultoni and P. minutus are genetically isolated from each other even though morphologically they are very similar, and in places sympatric. This genetic isolation was inferred from the presence of private alleles in some species, and from differences in their degrees of observed heterozygosity and in the observed frequencies of shared alleles in all the species examined. Temporal variations were not detected in P. newmani, P. acuspes and P. minutus. Populations of P. newmani and P. moultoni from the Atlantic and Pacific coasts of North America have apparently diverged. No such divergence was detected among samples of P. acuspes from Nova Scotia, the Canadian Arctic and the Baltic sea, and of P. minutus from Nova Scotia and Hudson Bay. P. elongatus collected in the English Channel differed from P. acuspes from the Baltic Sea.     

Genetic variability and level of differentiation in North Sea and Baltic Sea populations of the green alga<Emphasis Type="Italic"> Cladophora rupestris</Emphasis>

Cladophora rupestris is a perennial filamentous macroalga belonging to the Chlorophyta. It is widely distributed on both sides of the northern Atlantic Ocean and penetrates into the brackish Baltic Sea down to ca. 4 psu salinity. In this paper we present evidence for genetic differentiation of a Baltic form of this marine alga. We assessed genetic structure within and among 11 populations ranging along a salinity gradient from the Norwegian coast to the northern Baltic Sea proper. Samples of 328 individuals were studied using starch-gel protein electrophoresis to evaluate genetic variability and interpopulation differentiation based on allozymes. Of 11 loci examined, only one was polymorphic. For this locus, encoding superoxide dismutase (SOD-3), a total of seven alleles were distinguished. We found two genetically differentiated groups of populations of C. rupestris, one Baltic Sea group and one North Sea group, with a distinct border in the southern Kattegat near the entrance to the Baltic Sea. The genetic differentiation for SOD-3, expressed as pairwise F_ST values between the populations, was generally higher within the Baltic Sea group (0.10-0.43) than within the North Sea group (0.05-0.10); in the latter group also fewer pairs of populations differed significantly. Pairs of populations from different groups had the highest F_ST values (0.20-0.60). Hierarchical analysis of variance showed that 29.6% of the total variation in the SOD-3 locus was explained by variation between the two groups, while only 4.2% was explained by variation among the populations within the groups. The remaining variation (66.2%) was found within the populations.     

New insights on population genetic structure of <Emphasis Type="Italic">Delphinus delphis</Emphasis> from the northeast Atlantic and phylogenetic relationships within the genus inferred from two mitochondrial markers

The taxonomic status of common dolphins (Delphinus sp.) remains controversial despite the increased number of studies focusing on its populations. Two species are presently recognized, Delphinus delphis and D. capensis. Apart from a phylogeographic study of the genus Delphinus, genetic studies focusing specifically in the northeast (NE) Atlantic remain scarce. Following ecological and morphological evidence for the existence of different common dolphin morphotypes in the Portuguese coast, we examined the population structure of D. delphis from the NE Atlantic by comparing DNA sequences from two mitochondrial regions (control region and cytochrome b gene). Additionally, we compared the sequences obtained with existing sequences of D. delphis from the Azores, Black Sea, Canary Islands, Pacific Ocean, D. capensis and also two closely related delphinid species (Stenella coeruleoalba and Tursiops truncatus). In the analysis of the NE Atlantic populations, we found evidence for the existence of some level of genetic differentiation. In the broader phylogenetic analysis, D. delphis and D. capensis did not show reciprocal monophyly and we found a group of highly divergent individuals. We discuss the possibility for the existence of two divergent lineages that have evolved independently, a separate subspecies or events of introgressive hybridization. These findings could have important implications on a taxonomic level, although further investigation based on a larger geographical scale and on nuclear loci information will certainly elucidate the origin of these highly divergent individuals.     

Patterns of organic osmolytes in two marine bivalves, Macoma balthica, and Mytilus spp., along their European distribution

Patterns of nine intracellular free amino acids (FAA), which are utilized as organic osmolytes for salinity-induced cell volume regulation in marine osmoconformers, were compared in five Macoma balthica populations and seven Mytilus spp. populations along their European distribution. Three types of FAA patterns were classified within both taxa: a northern Baltic type, a southern Baltic type and an Atlantic/Mediterranean type which mainly differ regarding the share of alanine and taurine. Differences are discussed in relation to habitat salinity and population genetics. Along a salinity gradient, the total size of the intracellular FAA pool did not differ between sympatric M. balthica and Mytilus spp., and was significantly correlated with habitat osmolality in a range from 70 to 600 mmol kg⁻¹ H₂O (oligohaline to mesohaline) in both bivalves. In M. balthica, this correlation was mainly based on significant correlations of alanine (15–100 mmol kg⁻¹ DW), glycine (30–100 mmol kg⁻¹ DW) and taurine (0–70 mmol kg⁻¹ DW) with habitat osmolality. In Mytilus spp., only glycine (25–100 mmol kg⁻¹ DW) and taurine (4–180 mmol kg⁻¹ DW) were significantly correlated with habitat osmolality. The concentration of alanine was three times lower in Mytilus spp. than in M. balthica and did not correlate with habitat osmolality. Within a habitat osmolality range from 600 to 1,100 mmol kg⁻¹ H₂O (mesohaline to marine) the concentration of FAA remained constant in both taxa. It is suggested that under marine conditions additional organic osmolytes must become more important for cell volume regulation in Macoma and Mytilus.     

Population genetic structure of escolar (<Emphasis Type="Italic">Lepidocybium flavobrunneum</Emphasis>)

Escolar (Lepidocybium flavobrunneum) is a large, mesopelagic fish that inhabits tropical and temperate seas throughout the world, and is a common bycatch in pelagic longline fisheries that target tuna and swordfish. Few studies have explored the biology and natural history of escolar, and little is known regarding its population structure. To evaluate the genetic basis of population structure of escolar throughout their range, we surveyed genetic variation over an 806 base pair fragment of the mitochondrial control region. In total, 225 individuals from six geographically distant locations throughout the Atlantic (Gulf of Mexico, Brazil, South Africa) and Pacific (Ecuador, Hawaii, Australia) were analyzed. A neighbor-joining tree of haplotypes based on maximum likelihood distances revealed two highly divergent clades (δ = 4.85%) that were predominantly restricted to the Atlantic and Indo-Pacific ocean basins. All Atlantic clade individuals occurred in the Atlantic Ocean and all but four Pacific clade individuals were found in the Pacific Ocean. The four Atlantic escolar with Pacific clade haplotypes were found in the South Africa collection. The nuclear ITS-1 gene region of these four individuals was subsequently analyzed and compared to the ITS-1 gene region of four individuals from the South Africa collection with Atlantic clade haplotypes as well as four representative individuals each from the Atlantic and Pacific collections. The four South Africa escolar with Pacific mitochondrial control region haplotypes all had ITS-1 gene region sequences that clustered with the Pacific escolar, suggesting that they were recent migrants from the Indo-Pacific. Due to the high divergence and geographic separation of the Atlantic and Pacific clades, as well as reported morphological differences between Atlantic and Indo-Pacific specimens, consideration of the Atlantic and Indo-Pacific populations as separate species or subspecies may be warranted, though further study is necessary.     

Phylogeography of Cerastoderma glaucum (Bivalvia: Cardiidae) across Europe: a major break in the Eastern Mediterranean

Molecular variation of the lagoon cockle Cerastoderma glaucum (Poiret, 1789) was examined across the species range along European coasts, from the northern Baltic to the Black Sea and the Caspian Sea. A major phylogeographic break in mitochondrial COI gene sequences (divergence 6.2%) separated a group of Ponto-Caspian and Aegean Sea haplotypes from those to the west of the Peloponnese peninsula in the Mediterranean and in the Atlantic-Baltic sector. A similar subdivision, yet not entirely geographically coincident, was found at 16 allozyme loci (Nei's distance D=0.15). The results imply a long-term isolation of populations in parts of the Eastern Mediterranean or Black Sea basins through the Pleistocene. The subdivision does not concur with previous views on the systematics of C. glaucum complex, but the pattern is notably similar to that described earlier from some fish species. Marked phylogeographic structuring was also found at lower level within the major Mediterranean-Atlantic phylogroup of C. glaucum, which was divided into six regional or local haplotype subgroups. Divergence of these groups may date back one or several major Pleistocene climatic cycles. Local mtDNA diversity was particularly high in a sample from the Ionian Sea, whose mitochondrial identity was of the Mediterranean-Atlantic type, while its nuclear characters were more strongly associated with the Ponto-Caspian type. Patterns of shallower, star-phylogeny type diversity within the Ponto-Caspian phylogroup and in the Baltic Sea area may represent more recent, post-glacial generation of variation.     

Icy heritage: ecological evolution of the postglacial Baltic Sea reflected in the allozymes of a living fossil,the priapulid Halicryptus spinulosus

Genetic variation of 16 allozyme loci in 397 Halicryptus spinulosus (Priapulida) revealed overall polymorphism of P=0.438 and Hardy-Weinberg expectations for heterozygosity of H _e=0.060 for Baltic Sea stocks, H _e=0.143 for the White Sea and H _e=0.121 for Iceland. Maximal unbiased standard distances of D=0.0693 separated Baltic and White Sea populations. Nordic and Baltic populations could be distinguished by allozymes, but Baltic subsamples proved cohesive. Gene flow amounted to effective exchange values per generation of N _m=2.94 over 650 km of continuous habitat, N _m=10.65 over 175 km, and N _m=13.85 over 20 km. Gene flow started to decrease with geographic distance beyond a dispersal threshold of 20 km, but hierarchical G _ST-statistics indicated light isolation by distance beyond a minimum of 8 km. Gene flow is high for a benthic worm assumed to lack dispersal by pelagic larva, a paradox which cannot be resolved now. Baltic populations are characterized by lower heterozygosity than Nordic stocks. In the Baltic Basin, temporally continuous brackish-water conditions have only existed for the past 7000 years. The two possible colonization routes of H. spinulosus to the geologically young Baltic Sea imply genetic drift, whether by founder effect (sweepstake colonization from Iceland) or by refugial bottlenecking during the Ancylus phase of the Baltic Basin after a direct connection to the White Sea had been sequestered. Continued genetic drift is emphasized by lower heterozygosity in the ecologically unstable Belt Sea compared to the central Baltic. Allozymes falsify the reduced-mutability hypothesis to explain bradytelic evolution of Priapulida. Regional genetic homogeneity, ample polymorphism, and preference for anoxic black mud qualify H. spinulosus populations as indicators of microevolutionary responses to water circulation regimes or pollution in the Baltic Sea.     

The cockle <Emphasis Type="Italic">Cerastoderma edule</Emphasis> at Northeast Atlantic shores: genetic signatures of glacial refugia

The cockle Cerastoderma (Cardium) edule (L.) ranges from NW Africa to northern Scandinavia. Abundance in shallow coastal sediment is often high, and it attracts commercial harvest. In this study, a complex genetic pattern has been revealed by mitochondrial DNA in 383 individuals from 19 sampling sites. Parsimony network analysis of 79 haplotypes identified two dominant central haplotypes separated by low divergence. One is characteristic for a homogenous southwestern group of populations from Africa to the British Isles. The other is characteristic for a heterogeneous northern group with a deviant Arctic population. At the entrance of the Baltic Sea, a mixture zone of the dominant haplotypes was found. The estimated population expansion time for the northern haplotype group predates that of the southwestern one, suggesting northern glacial refugia and a subsequent southern expansion of C. edule populations.     

Regional patterns of microsatellite variation in <Emphasis Type="Italic">Mytilus galloprovincialis</Emphasis> from the Iberian Peninsula

The Almería-Oran Oceanographic Front (AOOF) has been proposed as an effective marine barrier to gene flow between the NE Atlantic Ocean and the Mediterranean Sea for several species. Previous studies using allozymes and mitochondrial DNA have reported a scenario of secondary intergradation between populations of Mytilus galloprovincialis from those basins, with the allelic frequencies of some loci showing abrupt clinal patterns across the AOOF. In this study, we aimed at testing the congruence between six neutral polymorphic microsatellites versus previous data on allozymes and mtDNA-RFLPs, at depicting the population structure of this species in the Iberian Peninsula. Microsatellite genotyping was scored on 17 samples of mussels collected in the Iberian coast, including some areas not sampled before. Microsatellites exhibited larger intrabasin diversity (F _SC = 1.72%, ), similar interbasin differentiation (F _CT = 2.81%) and fewer allelic clines than allozymes or mtDNA haplotypes. These results fully support the scenario of secondary intergradation with some ongoing gene flow between basins, as proposed in previous analyses. Moreover, this congruence between markers and analyses separated by a 12-year period (1988–2000) confirm the temporal stability of this marine barrier at shaping the Iberian phylogeographic break in M. galloprovincialis. In addition, the genetic continuity between the NE Atlantic (Portugal) and the Alboran Sea seems to be warranted across the Gulf of Cadiz and the Gibraltar strait after the present microsatellite data. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Availability of copper from phytoplankton and water for the bivalveMacoma balthica. II. Uptake and elimination from64Cu-labelled diatoms and water

The amount of copper taken up via algae and water byMacoma balthica from the Oosterschelde sea arm, S.W. Netherlands, was established using the radioisotope⁶⁴Cu. As far as we know, this isotope has never been used before in marine flod chain studies. As a model food source the marine diatomPhaeodactylum tricornutum was allowed to accumulate⁶⁴Cu for 1 d. These labelled algae were fed to the clams in the presence of the complexing agent EDTA (0.27mM). EDTA was added to prevent uptake of dissolved⁶⁴Cu that could be leaking from the labelled diatoms. In control experiments, unlabelled diatoms were fed toM. balthica in the presence of dissolved⁶⁴Cu (with and without EDTA) in order to assure a similar filtration activity. In repeated experiments with varying particulate/dissolved copper ratios, uptake through food always turned out to be at least as efficient as uptake from the water. It was concluded that Cu, associated with food, is well available for uptake byM. balthica.