Genetic stock structure of the swordfish (Xiphias gladius ) inferred by PCR-RFLP analysis of the mitochondrial DNA control region

Restriction fragment length polymorphism (RFLP) analysis was performed on PCR amplified DNA fragments containing the control region of the swordfish (Xiphias gladius Linnaeus, 1758) mitochondrial DNA. A total of 456 individuals comprising 13 local samples (six Pacific, three Atlantic, the Mediterranean Sea, two Indian Ocean and the Cape of Good Hope) were surveyed with four endonucleases (Alu I, Dde I, Hha I and Rsa I), yielding a total of 52 composite genotypes. Within-sample genotypic diversity (H) was high ranging from 0.702 to 0.962 with a value of 0.922 for the pooled sample. Significant geographic variation in the frequencies of genotypes and restriction patterns was revealed. The Mediterranean sample was highly distinct from all other samples. Further, Rsa I digestion revealed high levels of polymorphism in all but the Mediterranean samples, indicating that exogenous swordfishes rarely enter that body of water. Heterogeneity between the North and South Atlantic samples was significant, both of which differed from those of the Pacific. In contrast, the Indian Ocean samples were not significantly different from the samples of South Atlantic and Pacific. Genetic differentiation among the Pacific samples was low. The results indicate that the world-wide swordfish population is genetically structured not only among, but also within ocean basins and suggest that gene flow is restricted despite the absence of geographic barriers. Received: 28 August 1996 / Accepted: 2 October 1996     

Diel vertical migration of Sagitta setosa as inferred acoustically in the Black Sea

Swimming trajectories of chaetognaths Sagitta setosa Müller in the Black Sea were studied using an echosounder operating at 120 and 200 kHz and an acoustic Doppler current profiler (ADCP) operating at 150 kHz. S. setosa were acoustically discriminated with respect to vertical migration and swimming speed, according to dissolved oxygen (DO) concentration and the timing of migrations. S. setosa formed a concentration layer thicker than Calanus euxinus did (1–3 m). The migration was completed in about 2.5–4 h, upward migration starting before C. euxinus and downward migration after C. euxinus. Adult Sagitta swam fast only in the well-oxygenated layer (subsurface maximum DO). The DO was found to be a significant (p<0.05) variable by partial correlation between the speed and hydrographical parameters. This feature constituted an oxygen-dependent influence on S. setosa’s vertical swimming and distinguished S. setosa from C. euxinus. Chaetognaths migrated daily between the nearsurface and the oxycline or the suboxic zone (OMZ, see Fig. 3b for the layers characterized by DO). Whether the deepest depth limit of migration was the oxycline or the OMZ depended on the relative abundance of adult and immature (young) individuals in the concentration layer. In July and September, individuals belonging to a new generation did not migrate but stayed in subsurface water day and night.     

Population structure of loggerhead turtle (Carettacaretta) nesting colonies in the Atlantic and Mediterranean as inferred from mitochondrial DNA control region sequences

Mitochondrial (mt) DNA control region sequences were analyzed for 249 Atlantic and Mediterranean loggerhead turtles (Carettacaretta Linnaeus, 1758) to elucidate nesting population structure and phylogeographic patterns. Ten haplotypes were resolved among individuals sampled between 1987 and 1993, from ten major loggerhead nesting areas in the region. Two distinct phylogenetic lineages were distinguished, separated by an average of 5.1% sequence divergence. Haplotype frequency comparisons between pairs of populations showed significant differentiation between most regional nesting aggregates and revealed six demographically independent groups, corresponding to nesting beaches from: (1) North Carolina, South Carolina, Georgia and northeast Florida, USA; (2) southern Florida, USA; (3) northwest Florida, USA; (4) Quintana Roo, Mexico; (5) Bahia, Brazil; and (6) Peloponnesus Island, Greece. The distribution of mtDNA haplotypes is consistent with a natal homing scenario, in which nesting colonies separated by a few hundred kilometers represent isolated reproductive aggregates. However, a strong exception to this pattern was observed in the first group defined by mtDNA data (North Carolina to northeast Florida), which included samples from four nesting locations spread across thousands of kilometers of coastline. These locations were characterized by a single haplotype in 104 out of 105 samples, providing inadequate resolution of population divisions. In view of the subdivisions observed elsewhere, we attribute the lack of differentiation between North Carolina and northeast Florida to recent colonization of these warm temperate coastlines (after the Wisconsin glaciation) not to ongoing gene flow among spatially distinct nesting locations. The relationships among observed haplotypes suggest a biogeographic scenario defined by climate, natal homing, and rare dispersal events. The redefined relationships among nesting aggregations in the western Atlantic region (southeastern USA and adjacent Mexico) prompt a reconsideration of management strategies for nesting populations and corresponding habitats in this region. Received: 28 October 1996 / Accepted: 24 October 1997     

Extreme mitochondrial DNA divergence within populations of the deep-sea gastropod Frigidoalvania brychia

The deep sea supports a diverse and highly endemic invertebrate fauna, the origin of which remains obscure. Little is known about geographic variation in deep-sea organisms or the evolutionary processes that promote population-level differentiation and eventual speciation. Sequence variation at the 16 S rDNA locus was examined in formalin-preserved specimens of the common upper bathyal rissoid Frigidoalvania brychia (Verrill, 1884) to examine its population genetic structure. The specimens came from trawl samples taken over 30 years ago at depths of 457-1,102 m at stations in the Northwest Atlantic south of Woods Hole, Massachusetts, USA. Near the upper boundary of its bathymetric range (500 m), extremely divergent haplotypes comprising three phylogenetically distinct clades (average uncorrected sequence divergence among clades ~23%, ~3% within clades) were found at stations separated by a maximum distance of ~80 km, suggesting the presence of high levels of intraspecific divergence or the possibility of morphologically cryptic species. Only one of these clades was found at two stations in the mid- to lower part of F. brychia's depth distribution (800-1,100 m), suggesting lower clade diversity with increasing depth, although among-sample divergence, with a single exception, was minimal. One station was genetically divergent from all others sampled, containing a unique suite of haplotypes including two found only at this site. Steep vertical selective gradients, major oceanographic changes during the late Cenozoic, and habitat fragmentation by submarine canyons might have contributed to an upper bathyal region that is highly conducive to evolutionary change.     

Allozyme and mitochondrial DNA variation in yellowfin tuna (Thunnus albacares) from the Pacific Ocean

Samples of yellowfin tuna (Thunnus albacares) collected in 1991 and 1992 from the western, central and eastern regions of the Pacific Ocean were examined for genetic variability. Four polymorphic allozyme loci (ADA ^*, FH ^*, GPI-S ^* and GPI-F ^*) were examined in all samples and a fifth polymorphism (GDA ^*) was examined in western and central samples only. Samples were also screened for mitochondrial DNA variation following restriction analysis by two enzymes (BcII and EcoRI) detecting polymorphic cut sites. Eighteen mtDNA haplotypes were revealed, with an overall nucleon diversity of 0.678. A subset of individuals screened for eight restriction enzymes had an overall nucleon diversity of 0.724 and a mean nucleotide diversity per sample of 0.359%. No significant spatial heterogeneity was detected for alleles at the ADA ^*, FH ^*, GPI-S ^* and GDA ^* loci nor for the mtDNA haplotypes. Significant heterogeneity was detected for GPI-F ^*. At this locus, the two eastern samples (southern California and northern Mexico) were not significantly different from each other but were significantly different (P<0.001) from the five western/central samples (Philippines, Coral Sea, Kiribati, Hawaii-91 and Hawaii-92). GPI-F ^* 100 was the most common allele in western and central regions, GPI-F ^* 75 the most common in eastern samples.     

Introgression and mitochondrial DNA heteroplasmy in the Baltic populations of mussels Mytilus trossulus and M. edulis

A strong clinal change in salinity occurs between the Baltic Sea and the North Sea, Atlantic Ocean, in the Danish Straits, where hybridization zone between mussels Mytilus edulis and M. trossulus has been reported. Eleven samples of mussels were studied from the Danish Straits and the inner Baltic Sea. Extensive introgression of M. edulis alleles from the North Sea into populations throughout the Baltic was ascertained for mitochondrial DNA (mtDNA) and two nuclear markers (ME15–16 and ITS). In the opposite direction, introgression of M. trossulus alleles into the M. edulis background was observed at the EFbis nuclear marker in populations from Kattegat (Danish Straits). While only M. edulis F (female) mtDNA was present in the Baltic, there were still strong differences in frequencies in the control region length variants between the Danish Straits and the inner Baltic samples, and weaker variation in coding region ND2–COIII haplotype frequencies. In the assays of the two mtDNA regions, various patterns of heteroplasmy were detected in 32% of all the studied individual mussels; this includes the presence of distinct, independently inherited M and F mitochondria in males, as well as the presence of two different distinguishable F genomes. The male-inherited M mtDNA genomes are quite common in the mussels from the Danish Straits, but very rare in males from the inner Baltic. Instead, a recombined control region variant (1r), which seems to have taken over the role of the M genome, was present in a number of specimens in the Baltic. Observations of heteroplasmy for two F genomes in some females and males confirm disruptions of the doubly uniparental inheritance mechanism in the hybrid Baltic Mytilus.     

Growth history and inshore migration of the tropical eel, Anguilla marmorata, in the Pacific

A comparative study of the otolith microstructure and microchemistry of Anguilla marmorata glass eels in the western North Pacific (Japan, Taiwan, the Philippines, Indonesia) determined the timing of metamorphosis and age at recruitment to freshwater habitats with a view to learning about the early life history and recruitment of this species of tropical anguillid eel, which has a wide range throughout much of the western Pacific and parts of the Indian Ocean. Three new samples (from Japan, Taiwan, Indonesia) were analyzed and statistically compared along with two other previously published samples that were analyzed using the same techniques. Ages at metamorphosis and recruitment, respectively, were 123ᆡ.4 days (mean-SD) and 154ᆥ.0 days in specimens from Japan, 116ᆢ.6 days and 145ᆣ.6 days in those from Taiwan, 120ᆡ.0 days and 154ᆡ.5 days in the Philippines stock and 132Nj.7 days and 159ᆟ.7 days, and 120ᆣ.6 days and 152ᆣ.2 days in the Indonesian stock. The average duration of the period of metamorphosis estimated from otolith microstructure was very similar (15-17 days) in the specimens from all locations. A close linear relationship was found between the ages at metamorphosis and recruitment at all locations, suggesting that individuals that metamorphosed earlier were recruited to freshwater habitats at a younger age. Back-calculated hatching dates ranged over about 6 months of the year, suggesting that this species may spawn throughout much of the year. It is hypothesized that specimens from all four sites are from the same spawning population originating in a spawning area in the North Equatorial Current of the western North Pacific.     

Phylogeny and biogeographic history of hake (genus Merluccius), inferred from mitochondrial DNA control-region sequences

 Phylogenetic analyses of the left domain of the mitochondrial DNA control-region sequence have been used to examine the relationships among species of the genus Merluccius (Rafinesque, 1810), and to compare these with hypotheses based on morphological, meristic and allozyme characters. Analysis of aligned sequences revealed that transition bias was much lower than in mammalian mtDNA, and that nucleotide composition of control-region sequences was biased toward A and T. We have roughly calibrated a molecular clock for the genus, based on the rise of the Isthmus of Panamá, which is believed to have created a barrier to dispersal between marine species of the Atlantic and Pacific Oceans. Our mtDNA-based phylogeny was highly congruent with allozyme-based phylogenies, but poorly so with a previously described phylogeny based on morphology. Specifically, our phylogeny resolved two well-supported principal clades, one of American (west Atlantic and east Pacific) species and the other of Euro–African (east Atlantic) species. This suggests an evolutionary history during which the ancestral lineage of Merluccius was divided between two geographic regions, with subsequent dispersal and vicariant events resulting in the evolution and distribution of extant taxa. However, the relationships between some taxa within the American clade could not be resolved. We suggest that this is consistent with an hypothesis of a rapid origin and radiation of these taxa. Received: 12 December 1998 / Accepted: 15 October 1999     

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     

Mitochondrial control region variability and global population structure in the swordfish,Xiphias gladius

Little is known about what limits genetic exchange in highly vagile, open ocean vertebrate species, such as the swordfish Xiphias gladius L. Reduced abundance of swordfish in some regions, and increased fishing pressure in others, has raised concerns and fueled interest in a more complete evaluation of the resource. In this study, global population structure in swordfish was assessed by sequencing a 300 base pair segment of the 5 end of mitochondrial DNA control region from 159 swordfish collected in three ocean basins: the Mediterranean, Atlantic and Pacific, over the years 1988 to 1994. Among the 159 individuals, 95 polymorphic sites delineated 121 unique haplotypes, indicating a high level of polymorphism on a global scale. A phylogenetic analysis of the unique DNA haplotypes revealed two divergent clades with differing geographic distributions. Phylogeographic concordance of this pattern with that of two other pelagic fish species suggests a biogeographic explanation for this structure. An analysis of molecular variance (AMOVA) revealed significant geographic partitioning of molecular variation among the three ocean basins, indicating that swordfish populations are structured on a global scale. Estimates of genetic exchange among populations within an ocean basin were high, indicating panmixia within ocean basins. Since the haplotypic diversity exhibited by the swordfish control regions is extremely high, much larger sample sizes may be necessary to detect subdivision within ocean basins.     

Population structure of yellowfin tuna (Thunnus albacares) in the western Pacific Ocean, inferred from microsatellite loci

Five polymorphic microsatellite loci were examined in 1391 yellowfin tuna (Thunnus albacares) from eight regions of the western (Coral Sea, eastern Australia, Fiji, Indonesia, Philippines and Solomon Islands) and eastern (California and Mexico ) Pacific Ocean. Across all samples, numbers of alleles per locus ranged from 7 to 30 (mean: 17.0), and observed heterozygosities per locus ranged from 0.223 to 0.955 (mean: 0.593). Temporal collections were available for three areas: no significant temporal heterogeneity was observed for the Coral Sea (1991/1992 and 1995/1996 collections) or eastern Australia (1994/1995, 1995/1996, 1996/1997 and 1997/1998), but there was slight but significant heterogeneity at one locus (cmrTa-161) between the two Philippines collections (1994/1995 and 1996/1997). Genotypes generally showed a good fit to Hardy-Weinberg expectations within populations; only cmrTa-208 in the pooled Coral Sea population gave a significant deviation after Bonferroni correction for 40 tests, with a small but significant excess of homozygotes. Four loci showed no evidence of population differentiation following contingency Chi-squared and F_ST analyses. The fifth locus, cmrTa-161, showed small but significant differentiation (F_ST=0.002, P<0.001). This heterogeneity was largely a result of the Philippines 1994/1995 and Fiji collections; there was no correlation with geographic distance. The average F_ST across all five loci was very low (F_ST=0.002), but it was significant (P<0.001). It is unclear whether this low but significant differentiation reflects noise in the dataset, perhaps arising from experimental error, or real population differentiation. The finding of very limited population heterogeneity accords with most of the earlier allozyme and mitochondrial DNA studies of yellowfin tuna in the Pacific Ocean.     

Population structure of humpback whales in the western and central South Pacific Ocean as determined by vocal exchange among populations

For cetaceans, population structure is traditionally determined by molecular genetics or photographically identified individuals. Acoustic data, however, has provided information on movement and population structure with less effort and cost than traditional methods in an array of taxa. Male humpback whales (Megaptera novaeangliae) produce a continually evolving vocal sexual display, or song, that is similar among all males in a population. The rapid cultural transmission (the transfer of information or behavior between conspecifics through social learning) of different versions of this display between distinct but interconnected populations in the western and central South Pacific region presents a unique way to investigate population structure based on the movement dynamics of a song (acoustic) display. Using 11 years of data, we investigated an acoustically based population structure for the region by comparing stereotyped song sequences among populations and years. We used the Levenshtein distance technique to group previously defined populations into (vocally based) clusters based on the overall similarity of their song display in space and time. We identified the following distinct vocal clusters: western cluster, 1 population off eastern Australia; central cluster, populations around New Caledonia, Tonga, and American Samoa; and eastern region, either a single cluster or 2 clusters, one around the Cook Islands and the other off French Polynesia. These results are consistent with the hypothesis that each breeding aggregation represents a distinct population (each occupied a single, terminal node) in a metapopulation, similar to the current understanding of population structure based on genetic and photo‐identification studies. However, the central vocal cluster had higher levels of song‐sharing among populations than the other clusters, indicating that levels of vocal connectivity varied within the region. Our results demonstrate the utility and value of using culturally transmitted vocal patterns as a way of defining connectivity to infer population structure. We suggest vocal patterns be incorporated by the International Whaling Commission in conjunction with traditional methods in the assessment of structure.     

Co-existing populations of Pacific ocean perch, Sebastes alutus, in Queen Charlotte Sound, British Columbia

Variation at five microsatellite loci (Sal1, Sal2, Sal3, Sal4 and Sal5) was examined in approximately 1300 Pacific ocean perch (Sebastes alutus) sampled from 14 coastal sites in British Columbia, Canada. Mean observed heterozygosities by locus ranged from 71% to 88%, and by sample ranged from 75% to 84%. Theta values ranged from 0 to 0.04 over the five loci, and averaged 0.015. Among Pacific ocean perch samples, Š ranged from 0.001 to 0.056. Canonical discriminate analysis of multilocus genotypes and neighbour-joining analysis of pairwise genetic distances between samples both indicated the presence of three populations, one off the west coast of Vancouver Island (the Vancouver Island population) and two co-existing populations in Queen Charlotte Sound, Dixon Entrance and along the west coast of the Queen Charlotte Islands (the eastern and western QCI populations). Pacific ocean perch of the eastern and western QCI populations were caught in close proximity to each other, but individual samples showed little evidence of admixture. Fall and spring samples collected within geographic areas were genetically similar, indicating seasonally stable population structure. Restricted gene flow between the Vancouver Island and the two more northerly populations may result from limited adult dispersal and larval retention within the California Current and Alaska Gyre, respectively, but the presence of two populations within Queen Charlotte Sound cannot be explained entirely by larval retention hypotheses. The presence of two Pacific ocean perch populations in central British Columbia has implications for fisheries management.     

Reproductive cycles in two populations of the Pacific sand dollar Dendraster excentricus

East Pacific swimming crabs are poorly known ecologically. We trawled samples along 4 estuarine-continental shelf transects to provide information on the population structure and species composition of portunids along environmental stress gradients. Portunids comprised almost 99% of the benthic crabs. Callinectes toxotes dominates the freshest estuarine areas, but is replaced by C. arcuatus in estuary mouth and shallow-shelf samples. Deeper shelf samples are dominated by Euphylax robustus and Portunus sper. Great numbers of E. dovii, a pelagic species, were taken on the shelf; their size distribution implies intraspecific competition. Evidence is given that differences in salinity tolerance account for differential landward limits, but competition, predation by fishes, increased fouling and decreased available food exclude eurytopic species from physicochemically equable shelf waters.     

Population structure of the black tiger prawn, Penaeus monodon, among western Indian Ocean and western Pacific populations

We examined the population structure of the black tiger prawn, Penaeus monodon Fabricius, 1798, in the Indo-West Pacific by analyzing the geographic distribution of elongation factor 1-alpha intron sequences from specimens collected during the winter and spring of 1997. Both the molecular phylogeny of alleles and F-statistics indicated very strong differentiation between populations from the western Indian Ocean and western Pacific. This pattern is concordant with other recent studies of marine species in this region, implying that the Indo-Australian Archipelago represents a biogeographic break between populations in the Indo-West Pacific. F _ST-values among populations in the western Indian Ocean also indicate structure within this region, whereas no structure was found among western Pacific populations. Nucleotide diversity was significantly lower in the western Indian Ocean populations than in the western Pacific, implying that the populations have regional differences in demographic history. Received: 16 November 1998 / Accepted: 26 May 1999     

Allozyme and mitochondrial DNA variation in orange roughy,Hoplostethus atlanticus (Teleostei: Trachichthyidae): little differentiation between Australian and North Atlantic populations

Allozyme and mitochondrial DNA (mtDNA) genetic variation was compared in orange roughy (Hoplostethus atlanticus Collett) collected from waters off southern Australia and from waters about 22 000 km away in the North Atlantic west of Scotland. Samples were screened for 11 polymorphic allozyme loci and with 9 restriction enzymes. Significant heterogeneity between the two areas was detected for three allozyme loci (ADA ^*, CK ^* and GPI-1 ^*), and the overall G _ST (gene-diversity statistic) value of 1% was small but significant. Significant mtDNA haplotype heterogeneity was observed after ²- of haplotype frequencies but not after a G _ST analysis. Nucleotide sequence-diversity analysis showed very low net divergence (0.0023%) between the two samples. The Australian orange roughy had a lower allozyme heterozygosity and a lower mitochondrial DNA nucleon diversity than the North Atlantic sample. The very limited, although significant, allozyme and mitochondrial DNA heterogeneity between these areas suggests that there is some gene flow between these two populations. The species appears to be widespread, with its presence reported from the southern Pacific, southern Indian, and northern and southern Atlantic Oceans, and it is likely that gene flow between the antipodes is mediated by stepping-stone exchange between adjacent populations rather than by direct migration.     

Identification of genetic populations of the Pacific blue shrimp Penaeus stylirostris of the Gulf of California, Mexico

Species are often composed of discrete breeding units (i.e. populations or stocks) which, while not reproductively isolated from other such groups, may have limited opportunities to exchange genetic material because of geographic distance, barriers to migration, or spawning asynchrony. Low levels of gene flow between stocks may result, over time, in their genetic divergence, and species that are subdivided into morphologic or genetically distinct stocks are said to be structured. The aim of our investigation was to test whether or not Penaeus stylirostris from the Gulf of California (Mexico) was structured into genetically distinct populations. Shrimp samples were collected in 1996 from six regions of the Gulf where specimens with distinct morphologic characteristics had previously been identified. Statistical analysis of 324 RAPD loci (randomly amplified polymorphic DNA), resolved through polyacrylamide gels and scored for each of 78 specimens, permitted the quantification and comparison of between-stock genetic differences. The finding that genetically discrete stocks of P. stylirostris can be found in a small portion of the geographic distribution range of the species, disagrees with the long-held perception that this resource is panmictic in nature. This new evidence is not only of interest for selective breeding programs in the shrimp aquaculture industry, but is also relevant to the management of the Mexican shrimp fishery which, at present, is perceived and managed as a single stock. Received: 10 May 1999 / Accepted: 27 July 2000     

Close genetic similarity of Atlantic and Pacific skipjack tuna (Katsuwonus pelamis) demonstrated with restriction endonuclease analysis of mitochondrial DNA

Restriction endonuclease analysis of mitochondrial DNA indicated a surprisingly high degree of genetic similarity between skipjack tuna (Katsuwonus pelamis) from the Atlantic and Pacific Oceans. The present results (1983) support the findings of previous morphological and electrophoretic studies. Evidently, since the uplift of the Panamá land bridge about 3.1 million years ago, there has been continued genetic contact between Atlantic and Pacific skipjack tuna, presumably via the Southern Ocean.