Genetic analysis of tuna populations,<Emphasis Type="Italic"> Thunnus thynnus thynnus</Emphasis> and<Emphasis Type="Italic"> T</Emphasis>.<Emphasis Type="Italic"> alalunga</Emphasis>

The genetic population structures of Atlantic northern bluefin tuna ( Thunnus thynnus thynnus) and albacore ( T. alalunga) were examined using allozyme analysis. A total of 822 Atlantic northern bluefin tuna from 18 different samples (16 Mediterranean, 1 East Atlantic, 1 West Atlantic) and 188 albacore from 5 samples (4 Mediterranean, 1 East Atlantic) were surveyed for genetic variation in 37 loci. Polymorphism and heterozygosity reveal a moderate level of genetic variability, with only two highly polymorphic loci in both Atlantic northern bluefin tuna ( FH* and SOD- 1*) and albacore ( GPI- 3* and XDH*). The level of population differentiation found for Atlantic northern bluefin tuna and albacore fits the pattern that has generally been observed in tunas, with genetic differences on a broad rather than a more local scale. For Atlantic northern bluefin tuna, no spatial or temporal genetic heterogeneity was observed within the Mediterranean Sea or between the East Atlantic and Mediterranean, indicating the existence of a single genetic grouping on the eastern side of the Atlantic Ocean. Very limited genetic differentiation was found between West Atlantic and East Atlantic/Mediterranean northern bluefin tuna, mainly due to an inversion of SOD- 1* allele frequencies. Regarding albacore, no genetic heterogeneity was observed within the Mediterranean Sea or between Mediterranean and Azores samples, suggesting the existence of a single gene pool in this area.     

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.     

Phylogeography and evolution of the triplefin <Emphasis Type="Italic">Tripterygion delaisi</Emphasis> (Pisces,Blennioidei)

The genus Tripterygion (Risso 1826) is restricted to the eastern Atlantic and the Mediterranean, and comprises only three species. T. melanuros and T. tripteronotus are essentially endemic to the Mediterranean, while Tripterygion delaisi occurs in the Atlantic and in the Mediterranean. Two subspecies of T. delaisi have been described (T. d. xanthosoma in the Mediterranean and T. d. delaisi in the Atlantic). Several scenarios have been proposed for the evolution of T. delaisi subspecies, but so far its subspeciation process is not clear. In this study we present a population survey of T. delaisi including specimens from the two recognized subspecies. We combined a phylogeographic approach with estimates of the direction of migration (between the Atlantic and the Mediterranean) and of the coalescence time of the two subspecies, using polymorphic mitochondrial and nuclear genes. The results of this study clearly support the existence of two Tripterygion delaisi clades, one in the eastern Atlantic islands and another in the Atlantic coasts of Europe and in the Mediterranean. Historical migration between the islands and Western Europe plus Mediterranean was restricted, and showed a westbound trend, with a higher number of migrants going from the Western Europe plus Mediterranean into the islands. We estimated the time of coalescence of both groups of T. delaisi to be more recent than the onset of Pleistocene glaciations (1.7 Mya). Our results are consistent with previous hypothesis that consider successive dispersal events of a Tripterygion ancestor from the western African coast colonizing the Atlantic islands and the Mediterranean, promoting the evolutionary divergence between these areas.     

Present-day genetic composition suggests contrasting demographic histories of two dominant chaetognaths of the North-East Atlantic, <Emphasis Type="Italic">Sagitta elegans</Emphasis> and <Emphasis Type="Italic">S. setosa</Emphasis>

Sagitta elegans and S. setosa are the two dominant chaetognaths in the North-East (NE) Atlantic. They are closely related and have a similar ecology and life history, but differ in distributional ranges. Sagitta setosa is a typical neritic species occurring exclusively above shelf regions, whereas S. elegans is a more oceanic species with a widespread distribution. We hypothesised that neritic species, because of smaller and more fragmented populations, would have been more vulnerable to population bottlenecks resulting from range contractions during Pleistocene glaciations than oceanic species. To test this hypothesis we compared mitochondrial Cytochrome Oxidase II DNA sequences of S. elegans and S. setosa from sampling locations across the NE Atlantic. Both species displayed very high levels of genetic diversity with unique haplotypes for every sequenced individual and an approximately three times higher level of nucleotide diversity in S. elegans (0.061) compared to S. setosa (0.021). Sagitta setosa mitochondrial DNA (mtDNA) haplotypes produced a star-like phylogeny and a uni-modal mismatch distribution indicative of a bottleneck followed by population expansion. In contrast, S. elegans had a deeper mtDNA phylogeny and a multi-modal mismatch distribution as would be expected from a more stable population. Neutrality tests indicated that assumptions of the standard neutral model were violated for both species and results from the McDonald-Kreitman test suggested that selection played a role in the evolution of their mitochondrial DNA. Congruent with these results, both species had much smaller effective population sizes estimated from genetic data when compared to census population sizes estimated from abundance data, with a factor of ~10⁸–10⁹ difference. Assuming that selective effects are comparable for the two species, we conclude that the difference in genetic signature can only be explained by contrasting demographic histories. Our data are consistent with the hypothesis that in the NE Atlantic, the neritic S. setosa has been more severely affected by population bottlenecks resulting from Pleistocene range shifts than the more oceanic S. elegans.     

Phylogeographic differentiation of storm petrels (<Emphasis Type="Italic">Hydrobates pelagicus</Emphasis>) based on cytochrome<Emphasis Type="Italic"> b</Emphasis> mitochondrial DNA variation

We evaluated mitochondrial DNA (mtDNA) sequence variation in a 910 bp region of the cytochrome b gene of the storm petrel, Hydrobates pelagicus. Samples from birds collected from five populations in the North Atlantic Ocean and the Mediterranean Sea were investigated. Genetic differentiation within the Mediterranean basin was low but high in the Atlantic. Strong differences were noted between the Atlantic and the Mediterranean populations, confirming the distinction of the subspecies H. p. pelagicus and H. p. melitensis for the Atlantic and Mediterranean seabirds, respectively. Divergence between the two subspecies probably resulted from paleogeographic changes in the Strait of Gibraltar, which was likely the route used by H. pelagicus to invade the Mediterranean Sea. Current and past demography and ecology of the storm petrel is regarded as an explanation for the level of differentiation observed within each oceanic basin. We compare the phylogeographic pattern of the storm petrel to other seabirds that breed in the same regions.Communicated by S. A. Poulet, RoscoffC.C. and L.B. contributed equally to this paper.     

Genetic evidence fails to discriminate between <Emphasis Type="Italic">Macroramphosus gracilis</Emphasis> Lowe 1839 and <Emphasis Type="Italic">Macroramphosus</Emphasis><Emphasis Type="Italic">scolopax</Emphasis> Linnaeus 1758 in Portuguese waters

Fish belonging to the genus Macroramphosus are distributed throughout the Atlantic, Indian and Pacific oceans. Some authors consider this genus monotypic, Macroramphosus scolopax being the only valid species. Other authors consider (based on several morphological and ecological characters) that another species (Macroramphosus gracilis) exists and occurs frequently in sympatry with the first one. Intermediate forms are also reported in literature. In this paper, using the mitochondrial control region and the nuclear first S7 intron markers, we failed to find genetic differences between individuals considered to belong to both species as well as the intermediate forms. Our results suggest that in the northeastern Atlantic, Macroramphosus is represented by a single species, M. scolopax, with different morphotypes interbreeding in the sampling areas.     

<Emphasis Type="Italic">Patella aspera</Emphasis> and<Emphasis Type="Italic"> P. ulyssiponensis</Emphasis>: genetic evidence of speciation in the North-east Atlantic

The group of subspecies of Patella ulyssiponensis, described by Christiaens, was widely known as Patella aspera until recently. The group extends throughout the Mediterranean, on all Macaronesian islands, along the North African coast, and in Europe, as far north as southern Norway. Throughout its range it displays great variation in shell sculpture and colour. The aim of this work was to re-examine the various subspecies proposed by Christiaens and, genetically, to test the hypothesis that European continental populations belong to a different species than the one composed by Macaronesian (north-west African) populations. In the present work, this group was studied by allozyme electrophoresis of 21 loci and by six morphological variables. The monomorphic locus of malate dehydrogenase (Mdh-1) was found to be diagnostic for distinguishing European continental populations from those of north-west African archipelagos, confirming the specific status of both groups. The allele observed at this locus in the Macaronesian populations was novel, while European continental populations showed a plesiomorphic one, shared with all other north-west Atlantic patellids with the exception of Patina pellucida, suggesting a more recent origin of the Macaronesian species from the continental forms. Both species showed a genetic identity of 0.730±0.061, which allowed rough estimations of 6.5–3 Mya since speciation. The subspecies Patella ulyssiponensis deserta described by Christiaens was not confirmed by our genetic data. Other loci (glyceraldehyde-3-phosphate dehydrogenase and the second locus of lactate dehydrogenase) were partially diagnostic, in which both species showed different most common alleles. Morphologically, both species are easily recognised by shell characters and the results agree with previous findings, that continental populations are more homogeneous in shell morphology and radula characters than populations from the Macaronesia. This work supports retention of the earliest valid name, Patella ulyssiponensis Gmelin, with Lisbon, Portugal as type locality, for the European continental species, and Patella aspera Röding for the Macaronesian populations. Population subdivision within species was measured by theta, the estimator of F_st, showing in both P. aspera and P. ulyssiponensis a high degree of genetic structuring (=0.226 and 0.182, respectively) mostly explained by the large distances separating the populations within species.Communicated by J.P.Thorpe, Port Erin     

Evolution of coral reef fish<Emphasis Type="Italic"> Thalassoma</Emphasis> spp. (Labridae). 2. Evolution of the eastern Atlantic species

The genetic relationships within and among congeneric species of marine fish from the Atlantic and the Mediterranean are poorly known. Relationships among all five species of the wrasse genus Thalassoma present in the Atlantic and the Mediterranean were examined using sequence data from the mitochondrial control region. Sampling was focused on the mid-Atlantic T. sanctaehelenae (Valenciennes, 1839) and T. ascensionis (Quoy & Gaimard, 1834), the eastern Atlantic T. newtoni (Osório, 1891) from Sao Tome, and the eastern Atlantic/Mediterranean T. pavo (Linnaeus, 1758). Two western Atlantic species T. bifasciatum (Bloch, 1791) from the Caribbean and T. noronhanum (Boulenger, 1890) from Brazil served as outgroups. Tissues from a total of 132 individuals were sequenced. T. newtoni from Sao Tome preferentially grouped with the central Atlantic T. sanctaehelenae and T. ascensionis. T. pavo exhibits two distinct coloration patterns, one in the Cape Verde Islands and one in the eastern Atlantic Islands and Mediterranean. However, no genetic discontinuities between the Cape Verde Islands and the remaining samples or between Atlantic and Mediterranean individuals were found. Within Mediterranean populations of T. pavo, our data suggested the presence of a genetic break between eastern and western regions.Communicated by J.P. Grassle, New Brunswick     

Mitochondrial evolution and phylogeography in the hydrozoan<Emphasis Type="Italic"> Obelia geniculata</Emphasis> (Cnidaria)

The distribution and genetic structure of many marine invertebrates in the North Atlantic have been influenced by the Pleistocene glaciation, which caused local extinctions followed by recolonization in warmer periods. Mitochondrial DNA markers are typically used to reconstruct species histories. Here, two mitochondrial markers [16S rDNA and cytochrome c oxidase I (COI)] were used to study the evolution of the widely distributed hydrozoan Obelia geniculata (Linnaeus, 1758) from the North Atlantic and the Pacific and, more specifically, in the context of North Atlantic phylogeography. Samples were collected from six geographic localities between 1998 and 2002. Hydroids from the North Atlantic, North Pacific (Japan), and South Pacific (New Zealand) are reciprocally monophyletic and may represent cryptic species. Using portions of the 16S rDNA and COI genes and the date of the last trans-Arctic interchange (3.1–4.1 million years ago), the first calibrated rate of nucleotide substitutions in hydrozoans is presented. Whereas extremely low substitution rates have been reported in other cnidarians, mainly based on anthozoans, substitution rates in O. geniculata are comparable to other invertebrates. Despite a life history that ostensibly permits substantial dispersal, there is apparently considerable genetic differentiation in O. geniculata. Divergence estimates and the presence of unique haplotypes provide evidence for glacial refugia in Iceland and New Brunswick, Canada. A population in Massachusetts, USA, appears to represent a relatively recent colonization event.Communicated by J.P. Grassle, New Brunswick     

Population differentiation in megrim (<Emphasis Type="Italic">Lepidorhombus whiffiagonis</Emphasis>) and four spotted megrim (<Emphasis Type="Italic">Lepidorhombus boscii</Emphasis>) across Atlantic and Mediterranean waters and implications for wild stock management

Megrim, Lepidorhombus whiffiagonis, and four spot megrim, Lepidorhombus boscii, are two marine fish species of high commercial interest. Despite their quite heavy exploitation little is known on the genetic structure of their populations. The present work aimed at characterizing the first seven microsatellites markers available for the two megrim species. These new markers were in a second step employed to describe the population structure of the two species among their almost entire habitat range (Atlantic and Mediterranean samples). Our study confirmed the existence of a strong genetic difference between Atlantic and Mediterranean megrim species already described in the literature for L. whiffiagonis on the basis of variations at ribosomal genes. Additionally our analysis gave the first evidences of a strong genetic differentiation among Atlantic populations in both megrim species (within Atlantic global F_ST in L. whiffiagonis and L. boscii were respectively 0.158 and 0.145). When describing megrim population structure, the comparison between allele-frequency-based tests (F_ST comparisons) and genotype-based inferences (Bayesian approach) gave evidences of a hierarchical structure of the populations. In conclusion, our work enlighten the existence of two different stocks within the Atlantic Ocean and one in the Mediterranean Sea that will clearly need to be managed separately. As the present results do not fully support the current megrim stock boundaries they will surely help to rethink megrim management policies in the future.     

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.     

Regional differentiation and post-glacial expansion of the Atlantic silverside, <Emphasis Type="Italic">Menidia menidia,</Emphasis> an annual fish with high dispersal potential

The coastal marine environment of the Northwest Atlantic contains strong environmental gradients that create distinct marine biogeographic provinces by limiting dispersal, recruitment, and survival. This region has also been subjected to numerous Pleistocene glacial cycles, resulting in repeated extirpations and recolonizations in northern populations of marine organisms. In this study, we examined patterns of genetic structure and historical demography in the Atlantic silverside, Menidia menidia, an annual marine fish with high dispersal potential but with well-documented patterns of clinal phenotypic adaptation along the environmental gradients of the Northwest Atlantic. Contrary to previous studies indicating genetic homogeneity that should preclude regional adaptation, results demonstrate subtle but significant (F _ST = 0.07; P < 0.0001) genetic structure among three phylogeographic regions that partially correspond with biogeographic provinces, suggesting regional limits to gene flow. Tests for non-equilibrium population dynamics and latitudinal patterns in genetic diversity indicate northward population expansion from a single southern refugium following the last glacial maximum, suggesting that phylogeographic and phenotypic patterns have relatively recent origins. The recovery of phylogeographic structure and the partial correspondence of these regions to recognized biogeographic provinces suggest that the environmental gradients that shape biogeographic patterns in the Northwest Atlantic may also limit gene flow in M. menidia, creating phylogeographic structure and contributing to the creation of latitudinal phenotypic clines in this species.     

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 two species of <Emphasis Type="Italic">Lysidice</Emphasis> (Polychaeta,Eunicidae) associated to the seagrass <Emphasis Type="Italic">Posidonia oceanica</Emphasis> in the Mediterranean Sea

The aim of this work was to define the phylogeographic patterns of the two species of polychaete Eunicidae, Lysidice ninetta Audouin and Milne Edwards and Lysidice collaris Grube, both associated as sheath borers to the Mediterranean seagrass Posidonia oceanica, and with reference to their different origin and their actual geographic distribution. L. ninetta is distributed in the Atlantic and in the Mediterranean Sea while L. collaris is a tropical species, whose introduction into the Mediterranean Sea through the Suez Canal (lessepsian migrant) has been hypothesized in recent years. The two species have been often confused in the past, although they appear morphologically distinct. They share the same microhabitat (Posidonia sheaths) and they co-exist along a broad bathymetric range (1–30 m). Several populations for both taxa were sampled all along the coast of the Mediterranean basin. A variable no coding region of nuclear DNA (rDNA, ITS1) and a portion of a more conservative coding region of mitochondrial DNA (sub-unit one of citochrome oxidase, COI) were used as molecular markers. Both markers confirmed the separation between the two species. Low intraspecific polymorphism was present in L. collaris, together with absence of phylogeographic structure. In L. ninetta, instead, the presence of intraspecific cryptic lineages, sympatric in some sites, was recorded. Clustering of single populations in the two main clades was not always consistent between markers. The mitochondrial COI region showed more resolution at the given spatial scale. Our results suggest that Lysidice collaris could be recently introduced into the Mediterranean Sea from one or more separate events. On the other hand, for L. ninetta one could presume a re-colonization of the Mediterranean basin from the Atlantics, after the Messinian crisis (dry-out of the Mediterranean Sea, 5.5 my) with the subsequent separation of intraspecific lineages. The phylogeographic patterns of both Lysidice spp. are disjoined with respect to that of the host plant, P. oceanica. The obtained results suggest that environmental constraints and evolutionary history of these polychaetes and their host plant act in different ways to determine their actual genetic spatial structure.     

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.     

Life style and genetic variation in teleosts: the case of pelagic (<Emphasis Type="Italic">Aphia minuta</Emphasis>) and benthic (<Emphasis Type="Italic">Gobius niger</Emphasis>) gobies (Perciformes: Gobiidae)

The pattern of genetic variability of two species of Mediterranean gobiids was compared, with reference to their different life history traits (Aphia minuta paedomorphic and pelagic; Gobius niger metamorphosed and benthic). The aim was to evaluate how different life histories can affect the genetic structure in these marine teleosts. The study was carried out on populations of both species sampled in the western Mediterranean and in the Adriatic Sea. Seven restriction endonucleases were used for the RFLP analysis of a mitochondrial DNA segment comprising the NADH dehydrogenase subunits 3, 4L and 4. The results highlighted two different patterns of genetic variation, a weak genetic structure in A. minuta and population subdivision in G. niger. These observations may be explained not only in terms of the different dispersal capabilities of these species, but also considering that A. minuta is an abbreviate iteroparous spawner while G. niger is a protracted iteroparous spawner. Because abbreviate iteroparity is a reproductive strategy selected in stable environments with high resource availability, Pliocene and Pleistocene climate oscillations may have represented factors that negatively influenced the reproductive success of A. minuta, producing demographic fluctuations and bottlenecks, as suggested by the mismatch distribution analysis. The weak genetic structure of A. minuta populations seems to be therefore due to a more recent re-colonization of the Mediterranean basin after a severe population decline, rather than to the high vagility of this pelagic goby.     

Population structure of red porgy, <Emphasis Type="Italic">Pagrus pagrus</Emphasis>, in the Atlantic Ocean

The red porgy, Pagrus pagrus (L.), is a protogynous sparid associated with reefs and hard bottom habitat throughout the warm-temperate Atlantic Ocean. In this study, the degree of geographic population differentiation in Atlantic populations was examined with microsatellite and mitochondrial DNA markers (mtDNA). Six microsatellite loci were amplified and scored in 690 individuals from the eastern North Atlantic (Crete, Madeira, and Azores), western North Atlantic (North Carolina to Florida, and the eastern Gulf of Mexico), and Brazil. At two loci, fixed allelic differences were found among the three major geographic areas, while frequency differences were observed at three other loci. The DNA of 371 individuals was amplified at the mtDNA control region, and 526 bp were sequenced. Tamura–Nei’s D was used as a measure of nucleotide diversity and divergence: diversity averaged 0.011 within samples, while the corrected divergence averaged 0 between samples within the same area and 0.061 between samples from different areas. Transversion haplotype minimum spanning networks, nucleotide divergence, and F _ST values all show that the western Atlantic samples were more closely related to each other than any was to samples from the eastern North Atlantic. Within the western North Atlantic, no significant population differentiation was observed, and within the eastern North Atlantic, only the Azores sample showed detectable differences from Crete and Madeira. These data indicate general homogeneity within large areas, and deep divisions between these areas. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Genetic structure in lagoons: the effects of habitat discontinuity and low dispersal ability on populations of <Emphasis Type="Italic">Atherina boyeri</Emphasis>

Atherina boyeri is a common inhabitant of the Mediterranean brackish and coastal waters. Several recent studies proposed the recognition of three distinct forms/species (one lagoon/freshwater and two marine) within an A. boyeri complex. In this study, we used 10 newly available microsatellite loci to better define the genetic structure of the lagoon form. A total of 428 fishes from 11 lagoons along the Italian and Albanian coasts and three Italian lakes were analyzed. A pronounced genetic spatial structure was confirmed for all considered geographic scales (near, medium and broad). A restricted gene flow was detected, suggesting reduced population connectivity. The comparison of present data and previous studies, based on mtDNA and RAPD markers, allowed a preliminary identification of the roles of recent factors (gene flow, habitat discontinuity and water currents) and past events (e.g., Pleistocene glaciations) in shaping the pattern of genetic differentiation.     

Genetic diversity in two barnacle species, <Emphasis Type="Italic">Chthamalus stellatus</Emphasis> and <Emphasis Type="Italic">Tesseropora atlantica</Emphasis> (Crustacea,Cirripedia), with different larval dispersal modes in the archipelago of the Azores

ISSRs (Inter Simple Sequence Repeats) were employed to compare the genetic structure of Chthamalus stellatus and Tesseropora atlantica in the Azores. The two barnacles differ as regards life cycle: the former conducting pelagic life for about 22 days, the latter for only 24 h. Thirty individuals of each species were analysed from four locations of three Azorean islands. Five ISSR primers produced, respectively, 117 and 79 polymorphic bands in C. stellatus and T. atlantica. Estimates of among-samples genetic diversity suggested extensive connectivity in the former species and isolation in the latter. AMOVA further supported these results by attributing 2% of genetic variance to the among-islands component of C. stellatus and 30% to T. atlantica. Results are consistent with expectations based on species life history and corroborate the importance of pelagic stages in determining the degree of genetic structuring in benthic marine invertebrates.