Application of random amplified polymorphic DNA (RAPD) markers to evaluate intraspecific genetic variation in red mullet (Mullus barbatus)

The random amplified polymorphic DNA (RAPD) technique was used to evaluate genetic affinities among eight red mullet (Mullus barbatus L., 1758) samples from the Mediterranean Sea. Twenty-nine random primers were used. Despite the variability which was found within samples, no specific RAPD marker for the discrimination of the populations was detected. The data analysis revealed that the genetic diversity among populations is positively related to their geographic distances. The results of this study indicate that the estimated intraspecific variation may be more␣pronounced with RAPD markers than with allozymes when the two approaches are applied on the same populations. Received: 7 November 1997 / Accepted: 30 May 1998     

Allozyme variation in global populations of striped mullet, Mugil cephalus (Pisces: Mugilidae)

The striped mullet, Mugil cephalus Linnaeus, 1758, is one of the few species of marine shore fish with a worldwide circumtropical distribution. Because of this distribution and the dependency of M. cephalus on coastal waters during various phases of its life cycle, as well as nearshore living habits, questions have been raised regarding levels of genetic divergence and gene flow among transoceanic populations. To cast more light on this, allozyme variation at 27 presumptive gene loci was investigated in ten globally diverse populations. The observed heterozygosity ranged from 0.018 (Hawaii) to 0.081 (Florida), averaging 0.050. The proportion of polymorphic loci showed a similar trend. Several populations were characterised by fixed allelic differences. Estimated gene diversities were very high, the allele frequency variation among populations was found to be 68%; genetic distances reached 0.242, with an average of 0.117. Estimated rates of gene flow were high among Mediterranean populations (Nm = 7.26), and between Mediterranean and East Atlantic populations (Nm= 2.86), but extremely low between non-contiguous populations within the Indian, Pacific and Atlantic Oceans, where Nm ranged from 0.03 to 0.05. Received: 7 April 1997 / Accepted: 4 February 1998     

Polycyclic aromatic hydrocarbons in red mullet (Mullus bavbatus) and sediments from the Eastern Aegean Sea

The distribution of polycyclic aromatic hydrocarbons (PAHs) was investigated in red mullet (Mullus barbatus) and sediment samples in the eastern Aegean Sea. The data were collected during cruises in July and November 1994, in the framework of National Marine Measurement and Monitoring Programme in the Aegean Sea. PAHs were detected in the tissue of Mullus barbatus of different sex and size from nine sampling sites throughout the study area ranging from 0.03 to 0.46 ug/g (fresh weight). The observations show the high levels of PAHs were found in tissues of females. The average concentration of PAHs in Aegean Sea sediments ranged from 0.20 to 1.8 ug/g (dry weight). No significant correlation was observed between PAH and sedimentary organic carbon.     

Genetic structure and allozyme variation of sea bass (Dicentrarchus labrax and D. punctatus) in the Mediterranean Sea

This paper reports data on 28 allozyme loci in wild and artificially reared sea bass (Dicentrarchus labrax) samples, originating from either coastal lagoon or marine sites in the Mediterranean Sea. F _ST analysis (θ estimator) indicated strong genetic structuring among populations; around 34% of the overall genetic variation is due to interpopulation variation. Pairwise θ estimates showed that, on average, the degree of genetic structuring was much higher between marine populations than between samples from lagoons. Six polymorphic loci showed differences in allele frequencies between marine and lagoon samples. Multivariate analyses of individual allozymic profiles and of allele frequencies suggested that different arrays of genotypes prevail in lagoons compared to marine samples, particularly at those loci that, on the basis of previous acclimation experiments, had been implicated in adaptation to freshwater. On the other hand, variation at “neutral” allozyme loci reflects to a greater extent the geographic location of populations. Allozyme differentiation was also studied in a D. labrax population from the Portuguese coast. Average genetic distance between this population and the Mediterranean populations was quite high (Nei's D = 0.236) and calls into question the taxonomic status of the Portuguese population. Finally, genetic relationships between D. labrax and D. punctatus were evaluated. Average Nei's D was 0.648, revealing high genetic differentiation between the two species, even for two sympatric populations of these species in Egypt; thus gene flow was not indicated between species. Received: 24 October 1996 / Accepted: 27 November 1996     

Genetic divergence and phylogenetic inferences in five species of Mugilidae (Pisces: Perciformes)

Allozyme electrophoresis was used to compare the genetic divergence of global populations of Mugil cephalus Linnaeus, 1758 and two congeneric [M.␣curema Cuvier & Valenciennes, 1836; M. gyrans (Jordan & Gilbert, 1885)] and two more distantly related [Liza ramada (Risso, 1826); Xenomugil thoburni (Jordan & Starks, 1896)] species on the basis of 18 enzyme loci. The amount of genetic divergence among the species examined is in agreement with their present systematic status, the differences being larger among non-congeneric species than among species of the same genus. Intraspecific genetic distances in M. cephalus (average Nei's D= 0.154), although higher than those reported for conspecific populations of fish, appear to be small when compared to the interspecific values among mugilid species (0.821 ≤ Nei's D ≤ 1.744). Phylogenetic trees obtained by genetic distance methods and discrete character parsimony analysis were of similar topology, except for the relationships within the genus Mugil and for the arrangement of M. cephalus populations. Received: 7 April 1997 / Accepted: 4 February 1998     

Genetic variation in seven goby species (Perciformes: Gobiidae) assessed by electrophoresis and taxonomic inference

Data on the electrophoretic patterns of 20 enzyme loci were used to investigate relationships among seven Mediterranean goby species. The study evidenced different levels of genetic variability among the species. On the whole, the degrees of genetic differentiation were consistent with the current taxonomic recognition of species and genera. Phenetic and phylogenetic analyses agreed in clustering the samples in three groups, i.e. the Gobius (including Zosterisessor ophiocephalus), the Lesueurigobius species and Pomatoschiarua minutus. The taxa analysed probably evolved following the complex geological and climatic events that have been affecting the Mediterranean basin ever since the Miocene. Received: 20 June 1998 / Accepted: 5 February 1999     

Genetic structure of populations of two species of Chthamalus (Crustacea: Cirripedia) in the north-east Atlantic and Mediterranean

Protein electrophoresis on starch gels was used to investigate population genetic structure of the barnacles Chthamalus montagui Southward and C. stellatus (Poli) over their north-east Atlantic and Mediterranean ranges. In each species, a single locus exhibited marked differentiation of allele frequencies between Atlantic and Mediterranean localities; in C. stellatus, genetic differentiation between the two basins had not previously been noted. In both species, mean heterozygosity per locus appeared higher in the Mediterranean samples than in the Atlantic, and Mediterranean populations had more alleles at the loci studied. Possible explanations for the differentiation between the Atlantic and the Mediterranean populations are discussed. Received: 30 May 1996 / Accepted: 17 September 1996     

Is Syllis gracilis (Polychaeta: Syllidae) a species complex? An allozyme perspective

The genetic relationships between morphologically indistinguishable marine and brackish populations of Syllis gracilis Grube, 1840 (Polychaeta: Syllidae) were studied by means of allozyme electrophoresis. Samples of S. gracilis from marine coastal and brackish-water habitats were examined for variation at 13 presumptive loci. In addition, a sample of the closely related species S. prolifera (Krohn, 1852) was analysed. Five loci were multiallelic in at least one population of S. gracilis and eight loci in S. prolifera. Low to moderate levels of within-population genetic variability were found, with average expected heterozygosity values ranging from H = 0.068 (±0.043 SE) to 0.187 (±0.069 SE) in the populations of S. gracilis; higher values were found in S. prolifera (H = 0.325 ± 0.076). The presence of various private alleles indicated a marked genetic divergence among populations of S. gracilis, with Nei's genetic distances ranging from D = 0.000 to 0.833 and a highly significant F _ST value. Furthermore, evidence for strong genetic heterogeneity between two sympatric marine populations was found. UPGMA cluster analysis and multidimensional scaling pointed out a clear genetic divergence between brackish and marine populations. At least two genetically divergent entities occurred in marine and brackish habitats. This could be due to local adaptation of individuals coming from marine populations to brackish habitats, but more presumably to the occurrence of a species complex within S. gracilis. Received: 6 June 1999 / Accepted: 7 February 2000     

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     

Population structure of the common sole (<Emphasis Type="Italic">Solea solea</Emphasis>) in the Northeastern Atlantic and the Mediterranean Sea: revisiting the divide with EPIC markers

Spatial and temporal population genetic structures of the common sole, Solea solea, were studied in Northeastern Atlantic and Mediterranean Sea populations, using three polymorphic exon-primed intron-crossing (EPIC) markers. Results demonstrated significant multilocus differentiation among Eastern Mediterranean and a group composed by Western Mediterranean and Atlantic populations (θ = 0.150, P < 0.001), but also suggested unrecorded genetic differentiation of the Adriatic Sea population. No pattern of isolation-by-distance was recorded across the range covered by sampling, from the Kattegat to the Aegean Sea. Conversely to genetically structured Mediterranean populations, Atlantic populations ranging from Denmark to Portugal could be considered as representative of the same panmictic unit (θ = 0.009, not significant). Results further demonstrated stability of multilocus genetic structure among temporarily replicated cohort samples [0+, 1+, subadults] from several coastal and estuarine locations from Bay of Biscay, excepted for the amylase locus Am2B3-2 at one location (Pertuis d’Antioche). Despite coherence of such observed patterns of multilocus differentiation with previous allozymic surveys in sole, and with patterns generally obtained for other marine fish species, single-locus results from EPICs indicated divergent coalescence schemes supporting a complex response to ecology and history of sole’s populations. Results stress the use of nuclear genes such as EPIC markers to investigate population structure, but also historical, demographic, and possibly selective processes in marine fishes.     

Temporal stability and spatial divergence of mitochondrial DNA haplotype frequencies in red drum (Sciaenops ocellatus) from coastal regions of the western Atlantic Ocean and Gulf of Mexico

Restriction-site variation in mitochondrial (mt) DNA was assayed among 1675 red drum (Sciaenops ocellatus Linnaeus) sampled from 20 localities along the southeastern coast of the USA (western Atlantic) and the Gulf of Mexico (Gulf). Up to four consecutive year-classes (cohorts) were sampled at most localities. Nucleotide-sequence divergence among 170 mtDNA haplotypes identified ranged (in percentage) from 0.184 to 1.913, with a mean (±SD) of 0.887 ± 0.300. Comparisons of mtDNA haplotype frequencies across year-classes within localities were non-significant, indicating temporal stability of breeding components within localities. Significant heterogeneity in mtDNA haplotype frequencies was found across all localities, between (pooled) samples from the western Atlantic and the Gulf, and among geographically spaced, regional groupings in the Gulf. Genetic divergence between subpopulations of red drum in the western Atlantic and Gulf follows a pattern exhibited in other marine fishes, and probably stems from physical (historical environmental heterogeneity, absence of suitable habitat, and current patterns) and, perhaps, behavioral factors. Genetic differences among red drum in the Gulf appear to be due largely to an isolation-by-distance effect that is attributable to behavioral factors. The latter may include female philopatry to natal bays or estuaries, limited offshore (coastwise) movement of females relative to their natal bay or estuary, or both. Genetic divergence among red drum in the Gulf occurs despite high gene flow (estimated as the number of genetic effective migrants in an island mode). Conservation and management of red drum should be based on the premise that strategies for a given bay or estuary will impact geographically proximal bays or estuaries more than distal ones. Trajectories of correlograms in spatial autocorrelation analysis suggest a geographic neighborhood size, relative to genetic migration of red drum from a bay or estuary, of roughly 500 to 600 km. Received: 22 July 1998 / Accepted: 19 November 1998     

Allozyme and mitochondrial DNA variation in Cuban populations of the shrimp Farfantepenaeus notialis (Crustacea: Decapoda)

We investigated the genetic diversity among populations of the shrimp Farfantepenaeus notialis, the most abundant penaeid species around Cuba. A total of 25 allozyme loci were analyzed in samples of shrimps from seven localities at the south central platform of the island (Ana María Gulf). Samples from three of these localities and from Batabanó Gulf and Guacanayabo Gulf at the south west and south east platforms of the island, respectively, were also characterized at the mtDNA level through sequence variation of a 2027 bp segment including part of the COI and COIII genes. Of the 25 allozyme loci studied 9 were polymorphic: Akp2, Akp3, AmyB, Est3, Gdh, GP7, and Per1, 2 and 3. In contrast to mtDNA, the pattern of allozyme variation among localities revealed strong population structuring at Ana María Gulf, with significant F _st in all pairwise comparisons. The magnitude of F _st estimates as well as the grouping pattern obtained by a UPGMA analysis based on a distance matrix indicated that the level of differentiation was concordant with the geographical position of the localities and the hydrographic regime. Homogeneity of mtDNA suggested that differentiation of allozyme loci might be due to more recent events rather than historical isolation of the sampled populations. Ana María and Guacanayabo Gulf populations were differentiated by mtDNA from Batabanó Gulf, at the southwestern end of the island. The analysis showed three restriction site differences among them, suggesting genetic isolation of the two regions. The present results also suggest that an artificial introduction of larvae from Tunas de Zaza into Batabanó Gulf, in an effort to repopulate this fishing region, may have been ineffective. Received: 13 December 1999 / Accepted: 2 October 2000     

Sequence divergence of mitochondrial DNA indicates cryptic biodiversity in Octopus vulgaris and supports the taxonomic distinctiveness of Octopus mimus (Cephalopoda: Octopodidae)

DNA sequence diversity of octopods was investigated using the mitochondrial cytochrome oxidase III gene (mtCOIII). DNA was obtained from ethanol- or formalin-fixed tissue of 15 specimens belonging to Octopus mimus Gould, 1852, Octopus vulgaris Cuvier, 1797 and Scaeurgus unicirrhus d'Orbigny, 1840, from coastal waters of the Mediterranean (France), the southwestern Atlantic (Brazil), the Caribbean (Costa Rica) and the southeastern/tropical Pacific (north Chile/Costa Rica). A 612 bp fragment of the mtCOIII gene was sequenced and aligned to the orthologous sequences available from northeastern Pacific Octopus species. Possible phylogenetic relationships were reconstructed. The mtCOIII gene tree revealed two main clusters, one comprised O. rubescens, O. dofleini and O. californicus, while the other included all O. vulgaris specimens, O. bimaculatus, O. bimaculoides and O. mimus. With one exception all South American haplotypes including O. bimaculatus and O. bimaculoides appeared as the sister group of the Mediterranean haplotype of O. vulgaris, indicating that most of the South American O. vulgaris specimens investigated would not belong to the recently redescribed species O. vulgaris restricted to the Mediterranean and eastern Atlantic. The treatment of O. mimus as a species distinct from Mediterranean O. vulgaris is supported by a high nucleotide divergence of 12.7%. Based on the mtCOIII gene tree the existence of cryptic species among O. vulgaris-like octopods is suggested. Received: 15 January 1999 / Accepted: 15 October 1999     

High levels of gene flow in the surf bivalve Donax deltoides (Bivalvia: Donacidae) on the east coast of Australia

We surveyed patterns of allelic variation within twelve samples of the pipi Donax deltoides Lamarck from beaches separated by up to 1200 km but connected to varying degrees by the East Australian Current. We used these data to test the prediction that the irregular patterns of water movement would cause genetic differentiation in pipis, so that there would be more genetic variation within and among the more southern regions than the northern regions. We found that six loci were at least moderately variable within all samples, and there were no clear geographic patterns in allelic frequencies. In general, genotype frequencies within samples were consistent with predictions for an outcrossed, sexually reproducing species, and we detected no evidence of population subdivision. Within samples, with the exception of the peptidase loci, single-locus genotype frequencies were in close agreement with expectations for Hardy–Weinberg equilibrium. We observed no significant linkage disequilibrium for any pairwise comparison of loci in any sample. Our hierarchical analysis of genetic variation revealed little variation among all samples (F st = 0.009). Loci showed consistently low levels of subdivision (F st from 0.003 to 0.018). We found almost no variation among the four geographic regions sampled (F rt = 0.001). All variation was therefore attributable to variation among samples within regions (F sr = 0.010). These data imply that larvae are moving between regions and that levels of present or recent gene flow are high, and support the conclusions of other studies which have inferred widespread gene flow for animals dispersing via planktonic, outcrossed larvae in parts of this region. This implies that the East Australian Current is sufficient to produce strong larval connections despite its intermittent nature. If existing levels of population subdivision reflect current levels of gene flow, then these data imply that D. deltoides represents a single fishery on the east coast of Australia. Received: 16 September 1996 / Accepted: 25 September 1996     

Genetic relationships between marine and marginal-marine populations of Cerithium species from the Mediterranean Sea

Results of isozyme electrophoresis were used to explore the genetic relationships between several Mediterranean morphs of Cerithium (Gastropoda: Prosobranchia), for which taxonomy is currently uncertain because of high intraspecific variability and low interspecific differentiation. The large species, classically known as C. vulgatum Bruguière, 1789 was identified at four sites (two in the French Mediterranean and two in southern Spain). Two different larval types were found in the French sites, but poecilogony could not be demonstrated. Individuals collected from harbours were not genetically distinct from open-sea populations of classic C. vulgatum. However, a population in the Embiez lagoon (French Mediterranean) which morphologically resembles C. vulgatum did display distinct genetic traits, supporting its status as a separate species. Of the small Cerithium species usually known as C. rupestre, two sympatric species (C.“rupestre” Risso, 1826 and C. lividulum Risso, 1826) were distinguished. Genotype frequencies within the analysed populations revealed much heterozygote deficiency. F _ST values (fixation index measuring the effects of population subdivision) suggest a higher genetic differentiation for C. lividulum populations than for C. vulgatum populations. We assume that a high larval dispersal capability (via planktotrophy) allows a high gene flow between populations of C. vulgatum. Received: 24 November 1998 / Accepted: 24 September 1999     

Reproductive strategies of Mycalecontarenii (Porifera: Demospongiae)

The sexual and asexual phases of the reproductive cycle of the sponge Mycalecontarenii (Martens, 1824) were studied from samples collected over a period of 2 years (June 1994 to May 1996) in a Mediterranean coastal basin (Porto Cesareo, southwestern Apulia). The species is viviparous; year round it shows a discontinuous oocyte production. No males were found, possibly owing to the very short period of spermatogenesis. M.␣contarenii produces asexual buds during autumn and winter, and this process seems to be related to changes in water temperature. Even though, at present, we cannot evaluate the incidence of larval recruitment, we postulate that asexual reproduction in M.contarenii plays a crucial role in maintaining the sponge population. Received: 8 July 1997 / Accepted: 5 November 1997     

RAPD-PCR investigations of systematic relationships among four species of eels (Teleostei: Anguillidae), particularly Anguilla anguilla and A. rostrata

 We examined the genetic identities of 49 individuals of four species of eels in the genus Anguilla Shaw, A. anguilla, A. japonica, A. reinhardti and A. rostrata, using the random(ly) amplified polymorphic DNA (RAPD)-PCR technique. We used 15 random decamer primers. Out of the 22 148 amplification products detected, 454 markers were evaluated using Nei's distance coefficient, two numerical methods (neighbor-joining, UPGMA), and phylogenetic analysis using parsimony (PAUP). Both numerical and parsimony methods were congruent and confirmed the specific status of all four examined Anguilla species. In all dendrograms, all individuals of each of the species clustered together within four closed groups with highly significant bootstrap values (between 97 and 100%). Additionally 1 to 18 species-specific monomorphic diagnostic fragments were detected in three of the species; none were detected in A. anguilla. Mean interspecific genetic distances ranged from 0.384 to 0.559, with the minimum between A. anguilla and A. rostrata and the maximum between A. anguilla and A. reinhardti. Individuals collected at one locality and belonging to a single species did not cluster together, and neither were any diagnostic monomorphic fragments found for individuals of single localities. This probably reflects a random dispersal of larvae in ocean currents before they reach coastal waters. Among all possible species pairs, only the two Atlantic species were consistently clustered in all dendrograms, with highly significant bootstrap values (100%). Additionally, we detected ten diagnostic markers for this pair of eel species. From RAPD data, we suggest a phylogram which was routed by the descending analysis method: A. reinhardti appears to have been the first species to diverge from a putative common ancestor of the four eel species. Later, A. japonica separated from the hypothetical ancestor of the Atlantic species, which then split recently into A. anguilla and A. rostrata. The close relationship of A. anguilla and A. rostrata is indicated by their low genetic distance (GD = 0.384) and a high degree of shared RAPD fragments (SF = 71.2%); this compares with the following means for all other species pairs: ( = 0.531,  = 44.2%). The prerequisites for using RAPD data for the reconstruction of phylogenies are discussed. Received: 2 September 1999 / Accepted: 8 May 2000     

Life history of the red mullet Mullus surmuletus from the bottom-trawl fishery off the Island of Majorca (north-west Mediterranean)

The red mullet Mullus surmuletus is one of the main target species of the trawling fishery along the continental shelf off the Island of Majorca. The size distribution of the catches, and the reproduction, age and growth of this species have been studied based on sampling carried out from 1990 to 1992. The length range of the catches was between 10 and 32 cm, with a main distribution between 15 and 20 cm, but this included small specimens (recruits of 10 to 11 cm) from August. In the >19 cm length-class, females clearly dominated. Monthly variations in the gonadosomatic index (GSI) and in the percentage of mature specimens showed that males spawn from December to June, whereas the reproductive activity of the females centres around spring. Fifty percent of males and females mature at 15 and 16.8 cm, respectively, corresponding to 1 yr of age. Otolith age-readings indicate that the population exploited in the trawl fishery consists of six age-groups, including a very high proportion of individuals between 0 and 4 yr old. Population growth curves revealed that females grow comparatively slowly over a longer period of time and attain greater asymptotic sizes than males. The growth parameters for the whole population are: asymptotic length, L =31.28 cm; growth coefficient, K=0.211 yr^-1; theoretical age when length is zero, t ₀=-2.348 yr.     

Genetic differentiation in two sibling species of the brackish-water polychaete Hediste japonica complex (Nereididae)

Genetic divergence among ten populations of small- and large-egg forms of the brackish-water polychaete Hediste japonica complex was investigated on 14 isozyme loci by electrophoretic analysis. The two forms were distinguishable by complete allele substitutions at five loci, resulting in high genetic differentiation (Nei's D: 0.533 to 0.662). No genetic evidence of hybridization between the two forms was detected in sympatric populations in three rivers. These results indicate that the two forms are reproductively isolated, clearly showing that the two forms are distinct species. The genetic differentiation among populations was higher in the large-egg form (D: 0.005 to 0.111, G _ST: 0.435) than that in the small-egg form (D: 0.000 to 0.001, G _ST: 0.020). This genetic difference between the two forms seems to be attributable to a difference in their life histories. The average expected heterozygosity was low in populations of both the large-egg form (0.005 to 0.068) and the small-egg form (0.014 to 0.038) in comparison with other marine invertebrates. Received: 11 April 1997 / Accepted: 8 September 1997     

A molecular approach to the ecology of a mussel (Mytilus edulis – M. trossulus) hybrid zone on the east coast of Newfoundland, Canada

Mytilus trossulus Gould and M. edulis L. coexist in mixed populations in Atlantic Canadian waters. Although there is evidence that the two species hybridize in natural populations and that hybrids produce progeny through backcrosses, no study of the microgeographic distribution of the two forms and their hybrids has been made. Here we examine subtidal samples of mussels taken in July 1997 from two locations in eastern Newfoundland (Canada) and from wave-exposed and protected environments within each location. Shell lengths ranged from 15 to about 90 mm. Mussels were classified as pure forms or hybrids (F₁, F₂ and from backcrosses) based on four diagnostic markers, two allozyme loci (Mpi and Est-D) and two nuclear PCR-based DNA markers (ITS and Glu-5). In addition, a PCR-based mtDNA marker (COIII) was used to characterize the distribution of mtDNA mitotypes among pure and hybrid individuals. There were differences in the proportions of pure M. edulis and M. trossulus between sites and between environments at one location. M. trossulus was the predominant species at one of the two exposed sites. In all four samples, M. trossulus was also the predominant form among small individuals. The frequency of hybrids was 26% overall and did not differ among samples. Hybrids consisted mostly of backcrosses that were M. trossulus-biased among small mussels and M. edulis-biased among large ones. We conclude that both intrinsic genetic factors and extrinsic environmental factors influence the relative frequency of M. edulis, M. trossulus and their hybrids. Received: 29 June 1998 / Accepted: 4 November 1998