Evolution of population structure in an estuarine‐dependent marine fish

Restriction site‐associated DNA (RAD) sequencing was used to characterize neutral and adaptive genetic variation among geographic samples of red drum, Sciaenops ocellatus, an estuarine‐dependent fish found in coastal waters along the southeastern coast of the United States (Atlantic) and the northern Gulf of Mexico (Gulf). Analyses of neutral and outlier loci revealed three genetically distinct regional clusters: one in the Atlantic and two in the northern Gulf. Divergence in neutral loci indicated gradual genetic change and followed a linear pattern of isolation by distance. Divergence in outlier loci was at least an order of magnitude greater than divergence in neutral loci, and divergence between the regions in the Gulf was twice that of divergence between other regions. Discordance in patterns of genetic divergence between outlier and neutral loci is consistent with the hypothesis that the former reflects adaptive responses to environmental factors that vary on regional scales, while the latter largely reflects drift processes. Differences in basic habitat, initiated by glacial retreat and perpetuated by contemporary oceanic and atmospheric forces interacting with the geomorphology of the northern Gulf, followed by selection, appear to have led to reduced gene flow among red drum across the northern Gulf, reinforcing differences accrued during isolation and resulting in continued divergence across the genome. This same dynamic also may pertain to other coastal or nearshore fishes (18 species in 14 families) where genetically or morphologically defined sister taxa occur in the three regions.     

Fine-grained spatial genetic structure in the bivalve Gemma gemma from Maine and Virginia (USA), as revealed by Inter-Simple Sequence Repeat markers

Gemma gemma is a small ovoviviparous bivalve distributed in shallow sand flats along the North American Atlantic and Gulf of Mexico coasts. Genetic variation in G. gemma was analysed by means of Inter-Simple Sequence Repeats (ISSRs) at the following levels: (i) between localities (Maine and Virginia), (ii) among 10-m-diameter patches within localities, and (iii) within patches. Thirty individuals/patch and three patches/locality were analysed. Individuals were genotyped for 67 ISSR polymorphic loci from five primers. The portion of the genetic variation found between localities (2%) was small compared to that found either among patches within localities (37%) or within patches (61%). ISSRs in G. gemma allowed the detection of significant differentiation at individual and patch levels. By contrast, a low degree of genetic variability was found between localities. The small-scale genetic heterogeneity does not follow a simple, consistent pattern. Our results contrast with the generally accepted rule that aplanic species are locally homogeneous and globally heterogeneous and teleplanic species are the inverse.     

Spawning rhythms of common snook in Florida

Common snook Centropomus undecimalis were sampled monthly from the Jupiter–Lake Worth area of Florida's Atlantic coast during 1989 and 1991 (1452 fish) and from Tampa Bay on Florida's Gulf of Mexico coast during 1988 and 1989 (2090 fish). Group-synchronous oocyte development was demonstrated. Ovarian maturation began during March or April on both coasts. Spawning was first detected histologically in April during 1989 and 1991 on the Atlantic coast and during May in 1988 and in April in 1989 on the Gulf coast. In each year, spawning ended during October on the Atlantic coast and during September on the Gulf coast. Ovarian histological evidence suggested that individual females may spawn every 1·1–2·5 days between 1400 and about 2000 hours. Final oocyte maturation occurred independently of either tidal cycle or lunar phase, and some common snook were observed in prespawning or spawning condition on every day sampled. Spawning occurred in or near major inlets to the Atlantic Ocean and the Gulf of Mexico, in secondary passes to larger inland bays and bayous, and around nearshore islands.     

Temporal genetic variation of mitochondrial DNA and the female effective population size of red drum (Sciaenops ocellatus) in the northern Gulf of Mexico

We studied genetic drift of mitochondrial DNA (mtDNA) haplotype frequencies in a natural population of red drum (Sciaenops ocellatus) from the northern Gulf of Mexico (Gulf). The amount of genetic drift observed across temporally adjacent year classes (1986–89) was used to estimate variance effective (female) population size (N_ef). N_ef was estimated to be 14 308 and the ratio of female effective size to adult female census size was approximately 0.004, which is among the lowest value reported for vertebrate animals. Low effective size relative to census size among red drum in the northern Gulf may result from yearly fluctuations in the number of breeding females, high variance in female reproductive success, or both. Despite low genetic effective size relative to census size, the genetic effective population size of red drum in the northern Gulf appears sufficiently large to preclude potentially deleterious effects of inbreeding.     

Genetic effective size is three orders of magnitude smaller than adult census size in an abundant,Estuarine-dependent marine fish (Sciaenops ocellatus)

Using eight microsatellite loci and a variety of analytical methods, we estimated genetic effective size (N(e)) of an abundant and long-lived marine fish species, the red drum (Sciaenops ocellatus), in the northern Gulf of Mexico (Gulf). The ratio N(e)/N, where short-term variance N(e) was estimated via the temporal method from shifts in allele-frequency data over four cohorts and where N reflected a current estimate of adult census size in the northern Gulf, was approximately 0.001. In an idealized population, this ratio should approximate unity. The extraordinarily low value of N(e)/N appears to arise from high variance in individual reproductive success and perhaps more importantly from variance in productivity of critical spawning and nursery habitats located in spatially discrete bays and estuaries throughout the northern Gulf. An estimate of N(e) based on a coalescent approach, which measures long-term, inbreeding effective size, was four orders of magnitude lower than the estimate of current census size, suggesting that factors presently driving N(e)/N to low values among red drum in the northern Gulf may have operated similarly in the past. Models that predict N(e)/N exclusively from demographic and life-history features will seriously overestimate N(e) if variance in reproductive success and variance in productivity among spatially discrete demes is underestimated. Our results indicate that these variances, especially variance in productivity among demes, must be large for red drum. Moreover, our study indicates that vertebrate populations with enormous adult census numbers may still be at risk relative to decline and extinction from genetic factors.     

A modified stepping-stone model of population structure in red drum, Sciaenops ocellatus (Sciaenidae), from the northern Gulf of Mexico

Genetic studies of population or stock structure in exploited marine fishes typically are designed to determine whether geographic boundaries useful for conservation and management planning are identifiable. Implicit in many such studies is the notion that subpopulations or stocks, if they exist, have fixed territories with little or no gene exchange between them. Herein, we review our long-term genetic studies of red drum (Sciaenops ocellatus), an estuarine-dependent sciaenid fish in the Gulf of Mexico and western Atlantic Ocean. Significant differences in frequencies of mitochondrial DNA haplotypes and of alleles at nuclear-encoded microsatellites occur among red drum sampled across the northern Gulf of Mexico. The spatial distribution of the genetic variation, however, follows a pattern of isolation-by-distance consistent with the hypothesis that gene flow occurs among subpopulations and is an inverse (and continuous) function of geographic distance. However, successful reproduction and recruitment of red drum depend on estuarine habitats that have geographically discrete boundaries. We hypothesize that population structure in red drum follows a modified one-dimensional, linear stepping-stone model where gene exchange occurs primarily (but not exclusively) between adjacent bays and estuaries distributed linearly along the coastline. Gene flow does occur among estuaries that are not adjacent but probabilities of gene exchange decrease as a function of geographic distance. Implications of our hypothesis are discussed in terms of inferences drawn from patterns of isolation-by-distance and relative to conservation and management of estuarine-dependent species like red drum. Based on estimates of the ratio of genetic effective population size and census size in red drum, observed patterns of gene flow in red drum may play a significant role in recruitment.     

High genetic diversity, large inter-oceanic divergence and historical demography of the striped mullet

The striped mullet Mugil cephalus , L. is a circumtropical species whose extreme conservative morphology stands in contrast with the degree of genetic differentiation at a global scale. One hundred and fourteen mitochondrial control region DNA sequences were analysed from four localities in the Gulf of Mexico (Texas, Louisiana, Mississippi, Florida), one on the U.S.A. east coast (N. Carolina), and one in Hawaii, giving very high levels of molecular diversity ( h =1·0 every haplotype was unique in all samples, π=1·1–2·0% in Gulf–Atlantic and π=3·1% in Hawaii). With no genetic evidence of dramatic population expansions, mismatch distributions were still very different in each ocean. Stable population levels in Hawaii have fostered the generation and persistence of very high molecular diversity, but Gulf–Atlantic samples suggest a shorter span of population stability. Ninety-five per cent of the molecular variance was allocated between ocean basins and virtually none among Gulf–Atlantic samples. A neighbourjoining reconstruction revealed Atlantic- and Pacific-specific lineages separated by more than 24% uncorrected sequence divergence ( d =0·49 Tamura-Nei Gamma-corrected). The lack of phylogeographic structure among Gulf–Atlantic samples corroborated the AMOVA results and supported the existence of a single population with high levels of gene flow along the Gulf of Mexico and north-west Atlantic coasts. The genetic differentiation between oceans points to the absence of gene flow and an accelerated rate of mitochondrial evolution in the genus Mugil .     

Allozyme analyses of the genus Trisopterus: taxonomic status and population structure of the poor cod

Genetic analysis of the four Trisopterus (Gadidae) taxa suggests that the interrelationships of the two morphs of poor cod ( T. minutus minutus in the Atlantic and T. minutus capelanus in the Mediterranean) should be reconsidered. The Mediterranean poor cod T. m. capelanus is more closely related to bib T. luscus than to the Atlantic poor cod, so the population structure in the Atlantic and Mediterranean poor cod must be considered separately. Among 635 Atlantic individuals there was some evidence of poor cod population differentiation (allele frequency heterogeneity test P <0·0005; F _ST=0·0135, P ≤0·0005). Levels of genetic variation were similar to those reported for related gadoid species. Some differentiation was present on the Norwegian coast (samples from Trondheimsfjord) and between the Faeroe Islands (Faeroe Bank) and the adjacent European coastal location. In contrast no statistically significant population differentiation was evident in Mediterranean poor cod, but fewer samples and individuals were screened.     

Genetic variation and stock structure of school mackerel and spotted mackerel in northern Australian waters

The total mean sample heterozygosity calculated from eight polymorphic loci was 0·172 (0·047 S.E.) (F_st 0·025) for school mackerel Scomberomorus queenslandicus , and for spotted mackerel S. munroi was 0·110 (0·074 S.E.) (F_st 0·038). There was no evidence of temporal variation for either species as significant genetic differences were not detected between months or year classes within areas. Spatially, school mackerel have a complex stock structure, with stocks being associated with large embayments. In contrast, spotted mackerel appear to comprise a single stock in Australian east coast waters. Both species showed a significant pattern of stock structure between Australian east coast and northern (Arafura Sea) samples.     

Spatial variation in parasite abundance: evidence of geographical population structuring in southern garfish Hyporhamphus melanochir

Southern garfish Hyporhamphus melanochir were examined for metazoan parasites from nine sites in three regions (Spencer Gulf, Gulf St Vincent and northern Kangaroo Island) in South Australia to document parasite assemblages, identify candidate species suitable for use as biological tags and investigate spatial variation in parasite abundance. Four ectoparasite and 10 endoparasite species were identified representing Cestoda, Trematoda, Monogenea, Nematoda, Acanthocephala, Copepoda and Isopoda. Lernaeenicus hemirhamphi, Micracanthorhynchina hemirhamphi, Mothocya halei and Philometra sp. were suggested for 'permanent' biological markers. Multivariate discriminant function analysis showed that most sites could be distinguished based on differences in parasite abundance. Four endoparasites (Conohelmins sp., Hysterothylacium sp., M. hemirhamphi and Philometra sp.) were most important for site characterization. Limited spatial variation in permanent endoparasite abundance among localities in northern Spencer Gulf provided evidence for a distinct northern Spencer Gulf population with little interregional mixing. In contrast, considerable spatial variation in permanent endoparasite abundance between localities sampled off Kangaroo Island implied limited local movement and suggested H. melanochir may comprise a metapopulation structure. These results largely align with recent evidence from otolith chemistry that indicates fine-scale geographical population structuring in South Australian waters.     

Genetic isolation between Atlantic and Mediterranean albacore populations inferred from mitochondrial and nuclear DNA markers

Genetic population structure of Atlantic and Mediterranean albacore Thunnus alalunga was investigated using nucleotide sequence variations of the glucose‐6‐phosphate dehydrogenase gene intron ( G6PD ) and the mitochondrial DNA (mtDNA) D‐loop region ( Dloop ). Restriction analysis using Ase I digestion detected two major restriction types ( A and B ) at the Dloop locus with strong frequency differences between Atlantic and Mediterranean samples. Thirty‐six individuals of 100 Mediterranean albacore were of the B type whereas no B type individuals were found in the Atlantic samples ( n  = 102). Phylogenetic analysis using nucleotide sequence data of the Dloop locus indicated that the B type lineage recently arose from the ancestral A lineage in the Mediterranean Sea and has not dispersed into the Atlantic Ocean. The frequencies of two alleles ( L and S ) at the G6PD locus were significantly different between the samples from the Atlantic ( L  = 0·495) and the Mediterranean ( L  = 0·725), but no significant heterogeneity was observed between mtDNA‐ A and ‐ B types of the Mediterranean sample. These molecular data indicate that gene flow between the Atlantic and Mediterranean albacore populations have been considerably restricted and strongly suggest these populations should continue to be treated as two distinct management units.     

Post-ice age recolonization and differentiation of Fucus serratus L. (Phaeophyceae; Fucaceae) populations in Northern Europe

The seaweed Fucus serratus is hypothesized to have evolved in the North Atlantic and present populations are thought to reflect recolonization from a southern refugium since the last glacial maximum 18 000-20 000 years bp. We examined genetic structure across several spatial scales by analysing seven microsatellite loci in populations collected from 21 localities throughout the species' range. Spatial auto-correlation analysis of seven microsatellite loci revealed no evidence for spatial clustering of alleles on a scale of 100 m despite limited gamete dispersal in F. serratus of approximately 2 m from parental individuals. Pairwise theta analysis suggested that the minimal panmictic unit for F. serratus was between 0.5 and 2 km. Isolation by distance was significant along some contiguous coastlines. Population differentiation was strong within the Skagerrak-Kattegat-Baltic Seas (SKB) (global theta= 0.17) despite a short history of approximately 7500 years. A neighbour-joining tree based on Reynold's distances computed from the microsatellite data revealed a central assemblage of populations on the Brittany Peninsula surrounded by four well-supported clusters consisting of the SKB, the North Sea (Ireland, Helgoland), and two populations from the northern Spanish coast. Samples from Iceland and Nova Scotia were most closely aligned with northwest Sweden and Brittany, respectively. When sample sizes were standardized (N = 41), allelic diversity was twofold higher for Brittany populations than for populations to the north and threefold higher than southern populations. The Brittany region may be a refugium or a recolonized area, whereas the Spanish populations most likely reflect present-day edge populations that have undergone repeated bottlenecks as a consequence of thermally induced cycles of recolonization and extinction.     

Nuclear and mitochondrial DNA analyses reveal four genetically separated breeding units of the swordfish

Variants in the calmodulin gene intron 4 ( CaM ) and the mtDNA D-loop region ( D-loop ) detected by RFLP and nucleotide sequence analyses were used to investigate the global stock structure of the swordfish Xiphias gladius . Two alleles ( A and B ) were observed at the CaM locus among 8 samples (Mediterranean, Tarifa, four Atlantic areas, Indian and Pacific oceans) comprising 567 individuals. Genotype distributions at this locus within samples were in accordance with Hardy-Weinberg equilibrium. Significant differences in allele frequencies were observed between Mediterranean-Northwest Atlantic samples ( A =0·347–0·493) and those from tropical Atlantic and Indo-Pacific ( A =0·81–1). In the D-loop region, Alu I and Rsa I digestions detected 4 ( A - D ) and 8 ( A - H ) genotypes, respectively. The frequency of type C for Alu I was lower in Indo-Pacific samples (0·2) than Atlantic and Mediterranean samples (0·365–0·643). Frequency of type C for Rsa I increased from the Pacific (0·398) to the Northwest Atlantic (0·698) and Tarifa (0·949) samples, and the Mediterranean sample was fixed for type C . These samples were classified into three groups (Mediterranean-Tarifa, Atlantic and Indo-Pacific) using a heterogeneity test of mtDNA genotype distributions. Combined analysis of the CaM and D-loop loci indicated that there are at least four breeding units: Mediterranean, northwestern Atlantic, tropical to South Atlantic, and Indo-Pacific.     

Genetic differentiation in Chilean hake Merluccius gayi gayi (Pisces: Merlucciidae)

The genetic structure of Chilean hake Merluccius gayi gayi, was analyzed using starch gel protein electrophoresis. Samples were collected from four localities along the coast off Chile. A total of 1500 specimens sampled from Coquimbo, San Antonio, Talcahuano and Puerto Montt were used in the study. Genetic information was obtained for six allozyme loci Pgi-1, Pgi-2, Pgm, Idh-1, Idh-2 and Aat. The results of the analysis showed no significant allelic differences among samples from various localities, with an average F_ST value of 0.007 for all loci and samples. A heterogeneity test for all loci and specimens from the four localities showed only two significant values. Thus, Chilean hake populations along the coast of Chile appear to be genetically homogeneous.     

Microsatellite Variation Among Red Snapper (Lutjanus campechanus) from the Gulf of Mexico

Allelic variation at a total of 20 nuclear-encoded microsatellites was examined among adult red snapper (Lutjanus campechanus) sampled from 4 offshore localities in the Gulf of Mexico. The number of alleles at the 20 microsatellites ranged from 5 to 20; average (± SE) direct count heterozygosity values ranged from 0.148 ± 0.025 to 0.902 ± 0.008. No significant departures from expectations of Hardy-Weinberg equilibrium were found for any locus within samples, and genotypes at pairs of microsatellites appeared to be randomly associated, i.e., in genotypic equilibrium. Tests of homogeneity in allele distributions among the 4 localities were nonsignificant for 19 of the microsatellites. Allele distribution at microsatellite Lca 43 was heterogeneous among localities before (but not after) Bonferroni corrections for multiple tests executed simultaneously. Tests of homogeneity in the distribution of individual alleles at Lca 43 gave similar results: one low frequency allele was distributed heterogeneously among samples before, but not after, Bonferroni correction. Molecular analysis of variance indicated that more than 99% of variation at each microsatellite was distributed within sample localities. These results generally are consistent with the hypothesis of a single population (stock) of red snapper in the northern Gulf of Mexico. Received September 25, 2000; accepted January 16, 2001     

Disentangling complex genomic signals to understand population structure of an exploited,estuarine‐dependent flatfish

Interpreting contemporary patterns of population structure requires an understanding of the interactions among microevolutionary forces and past demographic events. Here, 4,122 SNP‐containing loci were used to assess structure in southern flounder (Paralichthys lethostigma) sampled across its range in the US Atlantic Ocean (Atlantic) and Gulf of Mexico (Gulf) and relationships among components of genomic variation and spatial and environmental variables were assessed across estuarine population samples in the Gulf. While hierarchical amova revealed significant heterogeneity within and between the Atlantic and Gulf, pairwise comparisons between samples within ocean basins demonstrated that all significant heterogeneity occurred within the Gulf. The distribution of Tajima''s D estimated at a genome‐wide scale differed significantly from equilibrium in all estuaries, with more negative values occurring in the Gulf. Components of genomic variation were significantly associated with environmental variables describing individual estuaries, and environment explained a larger component of variation than spatial proximity. Overall, results suggest that there is genetic spatial autocorrelation caused by shared larval sources for proximal nurseries (migration/drift), but that it is modified by environmentally driven differentiation (selection). This leads to conflicting signals in different parts of the genome and creates patterns of divergence that do not correspond to paradigms of strong local directional selection.     

Genetic Structure of Capelin (Mallotus villosus) in the Northwest Atlantic Ocean

Capelin (Mallotus villosus) is a commercially exploited, key forage-fish species found in the boreal waters of the North Pacific and North Atlantic Oceans. We examined the population structure of capelin throughout their range in the Canadian northwest Atlantic Ocean using genetic-based methods. Capelin collected at ten beach and five demersal spawning locations over the period 2002 through 2008 (N = 3,433 fish) were genotyped using six polymorphic microsatellite loci. Temporally distinct samples were identified at three beach spawning locations: Chance Cove, Little Lawn and Straitsview, Newfoundland. Four capelin stocks are assumed for fisheries management in the northwest Atlantic Ocean based on meristics, morphometrics, tag returns, and seasonal distribution patterns. Our results suggested groupings that were somewhat different than the assumed structure, and indicate at least seven genetically defined populations arising from two ancestral populations. The spatial mosaic of capelin from each of the two basal cluster groups explains much of the observed geographic variability amongst neighbouring samples. The genetic-defined populations were resolved at Jost’s D _est ≥ 0.01 and were composed of fish collected 1) in the Gulf of St. Lawrence, 2) along the south and east coasts of Newfoundland, 3) along coastal northern Newfoundland and southern Labrador, 4) along coastal northern Labrador, 5) near the Saguenay River, and at two nearshore demersal spawning sites, 6) one at Grebes Nest off Bellevue Beach on the east coast of Newfoundland, and 7) one off the coast of Labrador at Domino Run. Moreover, the offshore demersal spawners on the Scotian Shelf and Southeast Shoal appeared to be related to the inshore demersal spawners at Grebes Nest and in Domino Run and to beach spawners from the Gulf of St. Lawrence.     

Contemporary population structure and post‐glacial genetic demography in a migratory marine species,the blacknose shark,Carcharhinus acronotus

Patterns of population structure and historical genetic demography of blacknose sharks in the western North Atlantic Ocean were assessed using variation in nuclear‐encoded microsatellites and sequences of mitochondrial (mt)DNA. Significant heterogeneity and/or inferred barriers to gene flow, based on microsatellites and/or mtDNA, revealed the occurrence of five genetic populations localized to five geographic regions: the southeastern U.S Atlantic coast, the eastern Gulf of Mexico, the western Gulf of Mexico, Bay of Campeche in the southern Gulf of Mexico and the Bahamas. Pairwise estimates of genetic divergence between sharks in the Bahamas and those in all other localities were more than an order of magnitude higher than between pairwise comparisons involving the other localities. Demographic modelling indicated that sharks in all five regions diverged after the last glacial maximum and, except for the Bahamas, experienced post‐glacial, population expansion. The patterns of genetic variation also suggest that the southern Gulf of Mexico may have served as a glacial refuge and source for the expansion. Results of the study demonstrate that barriers to gene flow and historical genetic demography contributed to contemporary patterns of population structure in a coastal migratory species living in an otherwise continuous marine habitat. The results also indicate that for many marine species, failure to properly characterize barriers in terms of levels of contemporary gene flow could in part be due to inferences based solely on equilibrium assumptions. This could lead to erroneous conclusions regarding levels of connectivity in species of conservation concern.     

Genetic differentiation among populations of the shortfinned eel Anguilla australis from East Australia and New Zealand

Variability at seven microsatellite loci was used to survey the genetic population structure of the shortfinned eel Anguilla australis . Samples were collected from six estuaries along the east coast of Australia and from three estuaries around New Zealand. Hierarchical analysis of molecular variance of the five loci with good fit to Hardy–Weinberg genotypic proportions detected highly significant differences among samples ( F _ST= 0·016, P < 0·001). The fixation index between countries ( F _CT= 0·012, P < 0·001) was more than double the index among samples within countries ( F _SC= 0·005, P < 0·05). An unweighted pair-group method with arithmetic mean (UPGMA) tree also supported the separation of Australian and New Zealand populations, as did assignment tests, which correctly assigned 80 and 84% of the individuals to Australia and New Zealand, respectively. Isolation-by-distance appeared among samples overall ( r = 0·807, P < 0·001), but not among samples within countries ( r = 0·027, P > 0·05 in Australia; r = 0·762, P > 0·05 in New Zealand). These findings indicate that populations of A. australis in East Australia and in New Zealand may be reproductively isolated from one another. Genetic differentiation among populations of A. australis was two- to 10-fold higher than that among populations of other temperate eels in the North Atlantic Ocean, suggesting that two group of A. australis may reflect sub-species. Anguilla australis in the two countries have different genetic structures and thus require separate management. Genetic isolation between Australian and New Zealand populations indicates that juveniles recruit independently into these two regions from geographically or temporally isolated spawning areas.