Cryptic speciation in a high gene flow scenario in the oviparous marine sponge Chondrosia reniformis

Sponge systematics has been traditionally based on the study of the skeleton (spicules and spongin fibres). However, sponges of the genus Chondrosia are devoid of those skeletal features, making it difficult to distinguish between different species in the genus. Chondrosia reniformis Nardo, 1847, the type species of the genus, was described from the Mediterranean Sea. The lack of distinguishing morphological features may have been responsible for the widespread assignment of specimens of the genus to this species; as a result C. reniformis is considered to be a cosmopolitan species. In this work, populations of C. reniformis from the western Mediterranean (France) and the West Atlantic (Bermuda and Brazil) were analysed using allozyme electrophoresis for 13 enzyme loci. Levels of mean heterozygosity were high (Bermuda and Brazil H=0.27 and W Mediterranean H=0.12), as is often observed in sponge species. Gene identities observed between West Atlantic and Mediterranean populations were low (I=0.40-0.52, typical values for congeneric species), including the presence of four diagnostic loci. This level of divergence clearly shows that they are not conspecific. Hence, a worldwide or cosmopolitan distribution of C. reniformis would seem improbable. However, the West Atlantic samples (Bermuda and Brazil) were genetically similar (gene identity, I=0.88-0.95) over a distance of 8,000 km. This is the first report of genetic homogeneity in a sponge species over such a large geographical distance.     

Phylogeography of two squid (Loligo pealei and L. plei) in the Gulf of Mexico and northwestern Atlantic Ocean

The loliginid squids Loligo pealei LeSueur and L. plei Blaineville (both recently proposed for reclassification as Doryteuthis) are commercially important, similar in appearance, and sympatric throughout much of the northwestern Atlantic Ocean, the Gulf of Mexico, and the Caribbean Sea. To investigate possible cryptic speciation and population structure, we examined samples (collected from 1995 to 1997) of both species for restriction fragment length polymorphisms (RFLPs) in PCR products of the mitochondrial gene cytochrome c oxidase (subunit I). RFLP haplotypes were further characterized by direct sequencing. In North American waters, cryptic speciation was rejected by the far greater nucleotide sequence divergence between species (~14%) versus within species (<1%). Each species displayed about a dozen RFLP haplotypes, but only three of their respective haplotypes were found among 90% of L. pealei specimens (n=356) and 97% of L. plei specimens (n=431). For L. pealei, a genetic break existed between the northern Gulf of Mexico and the Atlantic Ocean; among sample units within each population, gene flow was consistent with panmixia. The phylogeography of L. pealei is likely a consequence of the eastward currents of the Florida Straits, the elevated temperatures of those surface waters, and the restriction of this species to the continental shelf. For L. plei, a genetic break existed between longitudes 88°W and 89°W, with the northwestern Gulf of Mexico and the northeastern Gulf-Atlantic Ocean comprising separate populations; among sample units within each population, gene flow fit an isolation-by-distance model. If the genetic break found for L. plei represents resident populations separated by nearshore physical parameters (e.g. effects of the Mississippi River and the sediment boundary at longitude 88°W), the lack of structure within the Gulf for L. pealei might be due to its distribution farther from shore. However, the two populations of L. plei probably represent annual recolonization from the southwestern Gulf of Mexico and from the eastern Caribbean Sea, whereas the populations of L. pealei probably are permanent residents within their respective regions.     

Global phylogeography of the fissiparous sea-star genus <Emphasis Type="Italic">Coscinasterias</Emphasis>

The fissiparous starfish genus Coscinasterias (Verrill) is represented in shallow waters around many of the world's continents. This wide distribution could be explained by dispersal, vicariance, or translocation associated with shipping, and represents an excellent system for marine biogeographic research. We conducted a global phylogeographic analysis of 42 Coscinasterias mtDNA cytochrome oxidase 1 sequences (15 haplotypes) from 18 sites, including representatives of all four recognised species. Phylogenetic analysis yielded a robust phylogeny, with strong support for the monophyly of the genus (90% bootstrap support) and of each separate species (99-100%). Haplotypes exhibited strong phylogeographic structure, with robust mtDNA clades often associated with distinct land masses. A general lack of genetic differentiation within sites may reflect fissiparity. However, shared haplotypes over larger distances (e.g. across Japan), and the presence of related haplotypes on adjacent land masses (e.g. Tasmania, New Zealand; 1.6-1.8%) suggest that long-distance dispersal is an important biogeographical process for Coscinasterias. The 4.0-4.4% divergence between Japanese and South African sister groups may relate to transequatorial dispersal around the onset of the Pleistocene. Divergent Atlantic and Mediterranean populations of C. tenuispina (maximum 1.5%) may deserve subspecific recognition, and high divergences within Australian C. muricata (maximum 8.0%) suggest a species complex.     

Effects of a complex archipelago on genetic subdivision of the intertidal limpet Siphonaria kurracheensis

Allozyme variation in the intertidal limpet Siphonaria kurracheensis was examined in 11 populations from the Houtman Abrolhos Islands, Western Australia, and four from the adjacent mainland, to test if the effects of these islands on genetic subdivision should include species with high dispersal. Consistent with extensive planktonic dispersal, genetic subdivision was low, with an average standardized variance in allelic frequencies (F_ST) of 0.010 over 400 km along the mainland, and 0.009 over 70 km in the Abrolhos Islands. Nevertheless, subdivision was statistically significant at the smallest scale examined (about 10 km), showing a pattern of local heterogeneity and large-scale homogeneity, which is common in planktonic dispersers. Among the island populations, heterozygosity was positively correlated with a multivariate index of exposure. The level of heterozygosity at the most exposed island sites is equivalent to that at the (also exposed) mainland sites, whereas it is lower in the most sheltered island sites. Although this indicates that some island populations are less well connected to the major pool of larvae, the overall effect of the archipelago on genetic subdivision is no greater than occurs on the mainland coast. This is contrary to findings in previously studied species with less potential for dispersal, suggesting that the local impediments to dispersal are selective in their effects, and are unlikely to be broadly important for genetic divergence in widely dispersing species.     

Sibling species or poecilogony in the polychaete<Emphasis Type="Italic"> Scoloplos armiger</Emphasis>?

In marine invertebrates multiple modes of development, or poecilogony, may occur in a single species. However, after close examination, many of such putative cases turned out to be sibling species. A case in point may be the cosmopolitan orbiniid polychaete Scoloplos armiger, which inhabits marine shallow sediments. In addition to the well-known direct, holobenthic development from egg cocoons, pelagic larvae have also been described. Our culture experiments revealed a spatially segregated source of the two developmental modes. All females of an intertidal population produced egg cocoons and no pelagic larvae. All but 2 out of 15 females of an adjacent subtidal population produced pelagic larvae and no egg cocoons. Based on these results we performed a molecular genetic analysis (RAPD-PCR) on three intertidal and four subtidal populations in the North Sea. Selected samples from all sites were analysed also by the AFLP method. We found significantly higher genetic diversity within subtidal than within intertidal populations. This is consistent with a wider dispersal by pelagic larvae and a smaller effective population size when development is holobenthic. Total genetic divergence is not related to distance but to the intertidal/subtidal division. We suggest that S. armiger actually represents two sibling species.     

Population genetics of the fissiparous holothurians Stichopus chloronotus and Holothuria atra (Aspidochirotida): a comparison between the Torres Strait and La Réunion

Collections of about 50 individuals from each of five populations of the fissiparous holothurian species Stichopus chloronotus and four populations of Holothuria atra were made in 1999. These populations were located in the Torres Strait (western Pacific) and La Réunion (western Indian Ocean). Allozyme electrophoretic surveys of five (S. chloronotus) and six (H. atra) loci were conducted to compare patterns of asexual reproduction and to investigate connectivity between regions separated by large geographic distances. Deviations from genotype frequencies expected under Hardy-Weinberg equilibrium, mostly heterozygote excesses, were observed in all populations of both species. The maximum contribution of sexual reproduction (calculated as the maximum number of sexually produced individuals: sample size=N*/N_i) was similar for all S. chloronotus (58-64%) and H. atra (76-92%) populations, and on the same level as previously reported for midshelf reefs of the Great Barrier Reef. The higher values in the latter species indicated greater contributions of asexual reproduction to S. chloronotus populations. Variability was strongly reduced in S. chloronotus populations at La Réunion, with only one locus being variable in that population. When the dataset was reduced to one representative per multi-locus genotype per population to reduce the effect of asexual reproduction on calculations on gene flow, F_ST values were not significantly different from zero, suggesting high gene flow between these regions. However UPGMA cluster analyses using Rogers' genetic distance, roughly clustered populations by region. In the case of H. atra, pooled populations within each region were significantly different from those of the other region. Thus, although some restrictions in gene flow and greater genetic distances between the regions may exist, those differences are distinctly less than those reported in previous studies on echinoderms over similar geographic scales. Despite the importance of asexual reproduction for the maintenance of local population size, this study also confirmed that the potential for widespread dispersal mediated by sexually produced larvae is large.     

Phylogeographic patterns of the mysid Mesopodopsis slabberi (Crustacea, Mysida) in Western Europe: evidence for high molecular diversity and cryptic speciation

The phylogeographic patterns among populations of Mesopodopsis slabberi (Crustacea, Mysida), an ecological important mysid species of marine and estuarine habitats, were analysed by means of DNA sequencing of a fragment of the mitochondrial cytochrome c oxidase subunit I (COI) and the 16S ribosomal RNA genes. Samples of M. slabberi collected from five Atlantic and two Western Mediterranean populations were investigated. Very high levels of within-population molecular diversity were observed in all samples (mean h=0.807 and π=0.0083), with exception of the Mediterranean Ebro population which contained only one haplotype. Differentiation among populations was high, and a clear phylogeographic break was observed between the Atlantic and Mediterranean populations. Moreover, a strong differentiation was detected between both populations in the Western Mediterranean Sea (Alicante and Ebro delta), while two divergent lineages occurred in sympatry within the Atlantic Mondego estuary. The high congruence between both the COI and 16S rRNA sequence data, the reciprocal monophyly of the different mitochondrial clades and the levels of nucleotide divergence between them suggest the presence of a complex of cryptic species within M. slabberi. Estimations of divergence time between the different mitochondrial lineages indicate that a split occurred during the late Miocene/early Pliocene. Such a divergence could be concordant with vicariant events during sea-level drops within the Mediterranean region at that time. However, within the Mediterranean Sea, the potential of divergence through ecological diversification cannot be ruled out.     

The genetic effects of larval dispersal depend on spatial scale and habitat characteristics

The intertidal gastropods Bembicium vittatum and Austrocochlea constricta, which have direct and planktonic larval development, respectively, occur sympatrically at sites across a number of islands at the Houtman Abrolhos archipelago and two harbours at Albany in Western Australia. Their distribution provide an opportunity to examine the effect of dispersal ability on levels of genetic subdivision at a number of spatial scales. F _ST (standardised variance in allelic frequencies) values in the range 0.361 to 0.396, determined from allozyme frequencies at 12 to 13 polymorphic loci, confirm isolation of Abrolhos and Albany populations, which are separated by 900 km of coastline, in both species. Within the Abrolhos and Albany, levels of subdivision in B. vittatum were high, but similar, as indicated by F _ST values of 0.091 and 0.090, respectively. In A. constricta, a mean value of 0.160 at the Abrolhos suggests severe restrictions to gene flow, while 0.021 at Albany indicates much stronger connections among populations. F _ST values at the Abrolhos support previous suggestions that this archipelago favours genetic subdivision in both direct and planktonic-developing species. The Albany harbours favoured subdivision only in B. vittatum, the low values of F _ST in A. constricta being attributed to strong mixing between the harbours, thus facilitating gene flow via planktonic larvae. The isolation of A. constricta populations at the Abrolhos can be explained in terms of highly localised recruitment, the result of limited water movement in complex intertidal habitats. The study illustrates the value of examining sympatric direct and planktonic developers in assessing the role of larval dispersal in patterns of genetic subdivision, and concludes that planktonic larvae may not promote gene flow over broad or even some fine spatial scales.     

Phylogeography of the sea star Marthasterias glacialis (Asteroidea, Echinodermata): deep genetic divergence between mitochondrial lineages in the north-western mediterranean

We explore the phylogeography of the broadcast spawner Marthasterias glacialis along south Europe and Azores. Sequences of the cytochrome c oxidase gene from 225 specimens, belonging to 10 localities, were analysed. We found 73 haplotypes grouped within two lineages (divergence 2.9%). One lineage was Atlanto–Mediterranean, whereas another one was exclusively Mediterranean. Estimation of lineages split goes back to 830,000–580,000 (±120,000) years ago. This suggests that sea-level oscillations during the Pleistocene glaciations promoted gene flow interruption, lineage divergence between basins and cryptic speciation. Secondary contact between populations allowed a recolonization of the Mediterranean by the Atlantic lineage. When animals of the Atlanto–Mediterranean lineage were considered separately, F _st index and AMOVA did not show significant differences between populations along either the Iberian Peninsula or basins. Isolation by distance between populations was not detected, and only populations of Plymouth and Azores showed significant differences to all the others. The remoteness of Azores islands might explain the structure of this population. Haphazard arrival of larvae and local extinctions rather than contemporary restricted gene flow might be responsible for the distinctive population structure of Plymouth.     

Population structure of red drum (Sciaenops ocellatus) in the northern Gulf of Mexico, as inferred from variation in nuclear-encoded microsatellites

Allelic variation at eight nuclear-encoded microsatellites was assayed among 967 red drum (Sciaenops ocellatus) sampled from four consecutive cohorts at seven geographic localities (=28 samples total) in the northern Gulf of Mexico (Gulf). Number of alleles per microsatellite ranged from 6 to 21; average direct-count heterozygosity values per sample (-SE) ranged from 0.560ǂ.018 to 0.903ǂ.009. Tests of Hardy-Weinberg equilibrium revealed significant departures from expected genotype proportions at one microsatellite, which was omitted from further analysis. Tests of genotypic equilibrium indicated that genotypes between pairs of microsatellites were randomly associated. Homogeneity tests of allele distributions across cohorts within localities were non-significant following correction for multiple tests executed simultaneously, and results from molecular analysis of variance indicated that the genetic variance component attributable to variation among cohorts did not differ significantly from zero. Homogeneity tests of allele distributions among localities (cohorts pooled) revealed significant differences both before and after correction for multiple tests. Neighbor-joining clustering of a pairwise matrix of Š values (an unbiased estimator of F_ST), spatial autocorrelations, and regression analysis revealed a pattern of isolation by distance, where genetic divergence among geographic samples increases with geographic distance between sample localities. The pattern and degree of temporal and spatial divergence in the nuclear-encoded microsatellites paralleled almost exactly those of mitochondrial (mt) DNA, as determined in a prior study. Stability of both microsatellite and mtDNA allele distributions within localities indicates that the small but significant genetic divergence among geographic samples represents true signal and that overlapping populations of red drum in the northern Gulf may be influenced by independent population dynamics. The degree of genetic divergence in microsatellites and mtDNA is virtually identical, indicating that genetic effective size of microsatellites and mtDNA in red drum are the same. This, in turn, suggests either that gene flow in red drum in the northern Gulf could be biased sexually or that red drum populations may not be in equilibrium between genetic drift and migration. If a sexual bias exists, the observation that divergence in mtDNA is considerably less than 4 times that of microsatellites could suggest female-mediated dispersal and/or male philopatry. The observed isolation-by-distance effect indicates a practical limit to dispersal. Approximate estimates of geographic neighborhood size suggest the limit is in the range 700-900 km. Although the genetic studies of red drum indicate significant genetic divergence across the northern Gulf, the genetic differences do not delimit specific populations or stocks with fixed geographic boundaries.     

Gene flow among vestimentiferan tube worm (Riftia pachyptila) populations from hydrothermal vents of the eastern Pacific

Allozyme data are presented for six discrete populations of the giant hydrothermal vent tube worm Riftia pachyptila Jones, 1981 collected throughout the species' known range along mid-ocean spreading ridges of the eastern Pacific Ocean. Contrary to an earlier report, levels of genetic variation are relatively high in this species. Estimates of gene flow based on F-statistics revealed that dispersal throughout the surveyed region is sufficiently high to counter random processes that would lead to losses of genetic diversity and significant population differentiation. R. pachyptila, like other species of tube worms, displays considerable morphologic variation among populations, but this diversity is not reflected in allozyme variation. Vestimentifera, in general, appear to show extensive phenotypic plasticity. In the light of the available genetic data, caution is warranted when making inferences about the taxonomic status of collections based on morphological variation alone. A general decrease in estimated rates of gene flow between geographically more distant populations supports the hypothesis that dispersal in this species follows a stepping-stone model, with exchange between neighboring populations in great excess of long-distance dispersal. High levels of gene flow have been recorded in a variety of vent fauna and may be a prerequisite for success of species found in the ephemeral habitats associated with regions of sea-floor hydrothermal activity.     

Genetic structuring and gene flow in Cerastoderma glaucum (Bivalvia: Cardiidae): evidence from allozyme variation at different geographic scales

Genetic variation at 16 enzyme loci was analysed in 20 Mediterranean and 1 Baltic population of Cerastoderma glaucum. Spatial genetic variation at different geographic scales was investigated. In general, this species was fairly genetically structured (over all loci Š=0.088), according to the fragmentary nature of its habitat. Spatial structuring of genetic diversity was shown to follow different models, depending on the geographic scale considered: a stepping-stone model provided a good fit at a wide scale, with gene flow inversely related to geographic distance, whereas at small scale, genetic relationships among samples could not be interpreted as simply the effect of physical distance among populations. Results are discussed taking into account the intrinsic and extrinsic factors which are most likely to affect the patterns of genetic structuring in coastal marine animals. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/10.1007/s00227-001-0753-x     

Morphological, reproductive, and genetic variability among three populations of Crucibulum quiriquinae (Gastropoda: Calyptraeidae) in northern Chile

Crucibulum quiriquinae (Lesson, 1830) is the only species of Crucibulum currently recognized in northern Chile. Recent analysis of three Crucibulum populations obtained in northern Chile demonstrates the existence of morphological, genetic, and reproductive differences among populations. Two populations present in Bahía Tongoy (30°15'S), one inhabiting the shells of the snail Turritella cingulata and the other shells of the pectinid Argopecten purpuratus, showed morphological differences. However, both had planktonic larval development and show low genetic divergence (D=0.002). A third population from Bahía La Herradura (29°58'S), which also inhabits the shells of T. cingulata, did not show morphological differences compared with its counterpart from Bahía Tongoy. However the Bahía La Herradura population had intracapsular development and metamorphosis, and a larger genetic distance (D=0.06) from both Tongoy populations. The results of the reproductive and genetic analyses strongly suggested that the two Tongoy populations, although showing morphological differences, are biologically the same species, Crucibulum quiriquinae, whereas the La Herradura population is a new species.     

Genetic structure of round scad mackerel <Emphasis Type="Italic">Decapterus macrosoma</Emphasis> (Carangidae) in the Indo-Malay archipelago

Round scad mackerel sampled in 1995-1998 were analysed for genetic variation using mitochondrial-DNA and nuclear-DNA markers. Sequence variation for a fragment of the cytochrome b gene (mitochondrial), amplified by polymerase chain reaction, was screened across individuals using single strand conformation polymorphism (SSCP). Sequence analysis of all SSCP haplotypes indicated two mitochondrial clades separated by, on average, 2.3% nucleotide divergence. The geographic distribution of haplotypes was homogeneous (Weir and Cockerham's [^(q)] \hat \theta =-0.002). Also, no geographic heterogeneity was detected for length polymorphism of Intron 1 of the gene encoding aldolase B ([^(q)] \hat \theta =0.005). Although homogeneity in allele frequencies throughout the Indo-Malay archipelago conformed to the expectations for a widely distributed pelagic fish in a highly connected habitat, this was at variance with the sharp geographic structure previously uncovered in Indian scad mackerel, Decapterus russelli, a fish with life-history characteristics similar to D. macrosoma. A remarkable similarity, however, was the occurrence of two similarly distinct clades within each species, suggesting a common history of geographic isolation. Low sea levels in the Pleistocene might have caused the separation and vicariance of populations within both D. macrosoma and D. russelli. Subsequent genetic exchange between populations would then have erased allele-frequency differences at the cytochrome b and aldolase B loci in D. macrosoma while some barrier to gene flow was maintained in D. russelli.     

Morphological and genetic adaptation to a lagoon environment: a case study in the bryozoan genus Alcyonidium

The Fleet (southern England) is a stable (ca. 5,000 years) coastal saline lagoon that supports a population of Alcyonidium resembling the common coastal epiphyte, Alcyonidium gelatinosum (L.). A combination of morphological, reproductive, and ecological characters was used to compare lagoonal and non-lagoonal proximate populations. Comparisons revealed a difference in the timing of spawning, considered to be related to the temporally restricted availability of viable substrata within the lagoonal basin. Allochronous spawning and spatial separation together suggest that the lagoonal taxon is reproductively isolated. The two populations were further compared with seven other coastal populations of Alcyonidium using randomly amplified polymorphic DNA (RAPD) analysis. The results confirm the individuality of the lagoonal taxon but also a close relationship with three A. gelatinosum populations. We present and consider four hypotheses that may account for the presence of this genetically distinct taxon: (1) diversification within the Fleet; (2) colonisation from another lagoon; (3) non-indigenous species status; and (4) introduction by shipping or other anthropogenically mediated dispersal mechanism. Significant diversification on the time scale involved has been demonstrated for isolated freshwater environments and, therefore, is feasible within a saline lagoon. Hypothesis 1 and, to a lesser extent, hypothesis 2 are consistent with the recognition of individual lagoons as 'biogeographic' islands of importance for their unique or characteristic biodiversity. The study also represents the first example of concordant morphological, reproductive, and genetic diversification in a marine bryozoan.     

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     

Social structures,genetic structures and dispersal strategies in Australian rabbit (<Emphasis Type="Italic">Oryctolagus cuniculus</Emphasis>) populations

In this study, the pattern of movement of young male and female rabbits and the genetic structures present in adult male and female populations in four habitats was examined. The level of philopatry in young animals was found to vary between 18-90% for males and 32-95% for females in different populations. It was skewed, with more males dispersing than females in some but not all populations. Analysis of allozyme data using spatial autocorrelation showed that adult females from the same social group, unlike males, were significantly related in four of the five populations studied. Changes in genetic structure and rate of dispersal were measured before and during the recovery of a population that was artificially reduced in size. There were changes in the rate and distance of dispersal with density and sex. Subadults of both sexes moved further in the first year post crash (low density) than in the following years. While the level of dispersal for females was lower than that of the males for the first 3 years, thereafter (high density) both sexes showed similar, low levels of dispersal (20%). The density at which young animals switch behaviour between dispersal and philopatry differed for males and females. The level of genetic structuring in adult females was high in the precrash population, reduced in the first year post crash and undetectable in the second year. Dispersal behaviour of rabbits both affects the genetic structure of the population and changes with conditions. Over a wide range of levels of philopatry, genetic structuring is present in the adult female , but not the male population. Consequently, though genetic structuring is present, it does not lead to inbreeding. More long-distance movements are found in low-density populations, even though vacant warrens are available near birth warrens. The distances moved decreased as density increased. Calculation of the effective population size (N_e) shows that changes in dispersal distance offset changes in density, so that N_e remains constant.     

Genetic population structure of the seagrass Thalassodendron ciliatum in sandy and rocky habitats in southern Mozambique

The genetic diversity of the dioecious seagrass Thalassodendron ciliatum in six populations in southern Mozambique was studied by means of random amplified polymorphic DNA (RAPD). Samples were taken from sandy and rocky habitats over a span of 880 km. Analysis of molecular variance (AMOVA) showed that most of the genetic variability (71.6% of total genetic variation) was observed within populations. There were also significant genetic differences among populations within each habitat (sandy and rocky). We did not find any significant overall genetic difference between sandy versus rocky populations, indicating that the morphological differences between plants from these two habitats are not maintained by reproductive isolation. There was no significant correlation between geographical and genetic distance, which is discussed in the light of current patterns along the coast. All sampled populations consisted of several genetically distinct individuals, indicating that sexual reproduction is widespread.     

Genetic evidence for the existence of cryptic species in an endangered clam <Emphasis Type="Italic">Coelomactra antiquata</Emphasis>

Coelomactra antiquata is a commercially important bivalve species, but has been suffering from severe population decline due to over-exploitation and the deterioration of environmental conditions. Previous genetic survey of C. antiquata conducted with allozymes combined with morphology revealed high levels of genetic differentiation between northern and southern populations which suggests a cryptic species might exist in C. antiquata. To test this hypothesis, amplified fragment length polymorphisms (AFLPs) and 16S rRNA gene sequence were used to re-evaluate the spatial genetic structure of six populations of C. antiquata along the coast of China. Both genetic markers display a sharp genetic break between the four northern populations (northern lineage) and two southern population (southern lineage). Large numbers of private alleles (AFLP) were found within the northern or southern populations and a deep divergence of about 6.5% in 16S rRNA gene sequence between the northern and southern lineages suggests the occurrence of potential cryptic or sibling species of C. antiquata. Applying previously published rates of mutation, divergence between the two lineages is estimated to have occurred approximately 3 million years ago and may be due to allopatric isolation during the middle Pliocene times. While no genetic differentiation was found within the northern or southern populations in both AFLP and 16S mtDNA markers, the results indicate that the northern and southern lineage should be managed separately and any translocation between the two areas should be avoided.     

Morphological and genetic evidence for vicariance and refugium in Atlantic and Gulf of Mexico populations of the hermit crab Pagurus longicarpus

The number and wide variety of southeastern United States marine taxa with significant differentiation between Gulf of Mexico and Atlantic Ocean populations suggests that these taxa may have experienced major vicariance events, whereby populations were subdivided by geological or ecological barriers. The present study compared variation in morphology, allozymes, and mtDNA in Gulf of Mexico and western Atlantic populations of the longwrist hermit crab Pagurus longicarpus Say collected during 1997 and 1998. Combined Atlantic populations had significantly fewer denticles on the second segment of the third maxilliped than did Gulf of Mexico populations, and the mean ratio of dactyl length to propodus length was significantly greater in the Atlantic crabs than in the Gulf of Mexico crabs. Allozyme allele frequencies at three loci showed genetic differentiation between a Gulf of Mexico population and two Atlantic populations. Analysis of mtDNA sequence data revealed a clear reciprocal monophyly between Gulf and Atlantic populations, with an estimated divergence age of ~0.6 million years ago. This estimated age of divergence is significantly more recent than an age previously estimated for its congener Pagurus pollicaris (~4 million years ago), suggesting that species with a similar genetic break between Gulf and Atlantic populations may not necessarily share an identical history. Surprisingly, there is evidence of geographic subdivision within Atlantic populations of P. longicarpus along the east coast of North America. This differentiation is especially strong between Nova Scotia and southern populations, suggesting that the Nova Scotia population may represent survivors from a northern refugium during the last glacial maximum.