Pleistocene diversification of the Pomacentrus coelestis species complex (Pisces: Pomacentridae): historical biogeography and species boundaries

Pleistocene eustatic changes in sea level are often invoked to explain genetic divergence among marine organisms. However, molecular phylogenies have revealed relatively few examples of speciation events dating to the Pleistocene. We present a species-level hypothesis of the timing of evolution for the Pomacentrus coelestis species complex (Pomacentridae), based on the nuclear S7 intron and the mitochondrial Cytb gene, and reconstruct ancestral range distributions across the timetree. Ancestral range reconstruction suggests the complex originated in the Coral Triangle and East Indian Ocean, with subsequent range expansion outward from this region. We suggest that land barriers to dispersal (e.g., Indo-Pacific barrier) may be responsible for the divergence between Indian (P. alleni, P. similis, P. caeruleopunctatus, and P. caeruleus) and Pacific (P. micronesicus, P. auriventris, and P. coelestis) species groups, and subsequent isolation by Pleistocene sea-level fluctuations in certain areas of the Coral Triangle (glacial refugia) may play an important role in the diversification of this species complex. Additionally, our analyses show cryptic lineages within P. micronesicus and highlight the need for comprehensive sampling within and among species to reveal recent speciation events.     

Calanus marshallae,a new species of calanoid copepod closely allied to the sibling species C. finmarchicus and C. glacialis

Species of the copepod genus Calanus frequently dominate the marine zooplankton in boreal and arctic waters. Up to now there have been no operational means of identifying several species closely related to Calanus finmarchicus. Reanalysis of these taxa, using material from plankton samples collected throughout Northern Hemisphere polar and coreal waters, shows that there are 3 sibling species which have been previously combined under the names C. finmarchicus and C. glacialis. Several new taxonomic characters permit unequivocal identification of C. finmarchicus, C. glacialis, and a new species, C. marshallae. Claims that C. finmarchicus and C. glacialis are subspecies are refuted; there is no evidence that the two species continuously intergrade either where they co-occur or where they are allopatric, nor is there evidence that the two species hybridize. C. finmarchicus is basically restricted to the North Atlantic Ocean and C. marshallae to the North Pacific Ocean and Bering Sea; C. glacialis is primarily an Arctic species, but its geographical distribution slightly overlaps those of the other two species. Taxa closely related to C. finmarchicus and C. helgolandicus probably represent two separate, but closely linked evolutionary lineage; species of these two lineages are placed in one of two species groups, the finmarchicus group and the helgolandicus group.     

Molecular phylogeny and population structure of tideland snails in the genus Cerithidea around Japan

Phylogenetic relationships and genetic population structures were analyzed for tideland gastropods in the genus Cerithidea around Japan on the basis of partial sequence of the mitochondrial COI gene. Large genetic divergence was shown between individuals of Cerithidea cingulata in the southern Ryukyus and those in the central Ryukyus and the Japanese Islands. Haplotypes of C. cingulata from the Japanese Islands were paraphyletic with the exclusion of a monophyletic group from the central Ryukyus. Genetic differentiation of C. cingulata was also detected between Amami-Oshima Island and Okinawajima Island. No genetic divergence was found between Cerithidea rhizophorarum in the Japanese Islands and its subspecies C. rhizophorarum morchii in the Ryukyu Islands. The lack of genetic divergence of Cerithidea largillierti between continental China and Japan suggests relatively recent migration between the Japanese Islands and the Asian continent. For all three Cerithidea species distributed in both the Japanese Islands and the Ryukyu Islands, the Tokara Gap and the Kerama Gap were shown to have acted as barriers to the dispersal.     

Hidden ancient diversification in the circumtropical lancelet Asymmetron lucayanum complex

The tropical lancelet Asymmetron lucayanum (= Epigonichthys lucayanus) is distributed from the western Indian Ocean to the central Pacific Ocean, and the western Atlantic Ocean. Molecular phylogenetic analysis of mitochondrial cytochrome c oxidase subunit I (COI) sequences (1,035 bp) of A. lucayanum (80 specimens from seven localities) showed clearly that this species is genetically distinguished into three major groups of geographical populations based on neighbor-joining tree using maximum likelihood distance (HKY model with invariable sites and gamma correction), suggesting the existence of three cryptic species. Our genetic data show that (1) inter-oceanic divergence time between Clade B (the West-Central Pacific) and Clade C (the Atlantic) (d = 6.6%, ca. 12 million years ago) was smaller than intra-oceanic divergence time between Clade A (the Indo-West Pacific) and Clade B (d=39.5%, ca. 100 million years ago); (2) there are two cryptic species in the West Pacific in sympatry; and (3) high gene flow is implied between the Maldives and the Ryukyus in Clade A (10,000 km distance), the Philippines and Hawaii in Clade B (8,500 km distance), and Barbados and Bermuda in Clade C (2,200 km distance).     

Molecular phylogeny and evolution of the pelagic copepod genus Neocalanus (Crustacea: Copepoda)

Portions of the mitochondrial genome (ca. 4 kb), encoding three protein-coding (COI, ND4L, ND6) and two ribosomal RNA (srRNA, lrRNA) genes, were sequenced for all six currently recognized species, plus one form, of the pelagic calanoid copepod genus Neocalanus. In Neocalanus gracilis, the ND6 gene was not found in the sequenced portion of the mitochondrial genome. Unambiguously aligned sequences were subjected to Bayesian, maximum-likelihood, maximum-parsimony, and neighbor-joining analyses using Eucalanus bungii as an outgroup. The resultant tree topologies from these four methods were congruent, robust, and all nodes were supported by high bootstrap values and posterior probabilities of 92–100%. Two tropical and subtropical species (N. gracilis and N. robustior) occupied the most basal position, and a subantarctic (N. tonsus) and three subarctic Pacific species (N. cristatus, N. plumchrus, and N. flemingeri) diverged subsequently. Transequatorial dispersal of the ancestral population during glaciations is suggested for this pattern of speciation, in which sister clades exhibited antitropical distributions. Although the area of ocean is much broader in the subantarctic than the subarctic Pacific, a higher number of species occur in the subarctic Pacific (three) than the subantarctic (one). The possibility that marginal seas, such as Japan Sea and Okhotsk Sea, function as natal areas for the divergence of species is discussed.     

Molecular population structure of the kuruma shrimp <Emphasis Type="Italic">Penaeus</Emphasis><Emphasis Type="Italic"> japonicus</Emphasis> species complex in western Pacific

In a previous study on the kuruma shrimp Penaeus japonicus from the South China Sea, we detected high genetic divergence between two morphologically similar varieties (I and II) with distinct color banding patterns on the carapace, indicating the occurrence of cryptic species. In the present study, we clarify the geographical distribution of the two varieties in the western Pacific by investigating the genetic differentiation of the shrimp from ten localities. Two Mediterranean populations are also included for comparison. Based on the mitochondrial DNA sequence data, the shrimps are separated into two distinct clades representing the two varieties. Variety I comprises populations from Japan and China (including Taiwan), while variety II consists of populations from Southeast Asia (Vietnam, Singapore and the Philippines), Australia and the Mediterranean. Population differentiation is evident in variety II, as supported by restriction profiles of two mitochondrial markers and analysis of two microsatellite loci. The Australian population is genetically diverged from the others, whereas the Southeast Asian and Mediterranean populations show a close genetic relationship. Variety I does not occur in these three localities, while a small proportion of variety II is found along the northern coast of the South China Sea and Taiwan, which constitute the sympatric zone of the two varieties. The present study reveals high genetic diversity of P. japonicus. Further studies on the genetic structure of this species complex, particularly the populations in the Indian Ocean and Mediterranean, are needed not only to understand the evolutionary history of the shrimp, but also to improve the knowledge-based fishery management and aquaculture development programs of this important biological resource.     

<Emphasis Type="Italic">Alexandrium catenella</Emphasis> (Dinophyceae), a toxic ribotype expanding in the NW Mediterranean Sea

The presence of the paralytic shellfish poisoning (PSP) dinoflagellate Alexandrium catenella in the north western (NW) Mediterranean Sea has been known since 1983. From this date on, the species has spread along the Spanish and Italian coastlines. Information concerning A. catenella isolates in the NW Mediterranean Sea was gained through phylogenetic studies. Twenty established toxic cultures of A. catenella taken from various NW Mediterranean Sea locations were analysed by nucleotide sequencing of the 5.8S rDNA and internal transcribed spacer regions. These rDNA ribosomal markers resulted useful in delineating the phylogenetic position of this species in the genus Alexandrium as well as in determining relationships between A. catenella isolates from different geographic areas. The phylogenetic position of the Mediterranean A. catenella ribotype, when compared to the “Alexandrium tamarense/catenella/fundyense species complex”, fits this species complex well. All the Mediterranean A. catenella isolates were constituted by only one genetic ribotype. By comparing the isolate sequences with those of other geographic areas, it revealed that the Mediterranean A. catenella ribotype was closely related to the A. catenella from Japan, Western Pacific Ocean. It was also evident that in temperate Japanese waters, a genetic variability was detected within A. catenella isolates; in fact, all strains resulted divergent showing as many as 15 mutational steps. The possibility that A. catenella has been recently introduced into the Mediterranean basin from temperate Asian areas is discussed.     

Genetic structure of wild European populations of the invasive Pacific oyster Crassostrea gigas due to aquaculture practices

As a result of aquaculture activities, Pacific oysters Crassostrea gigas (Thunberg, 1793) have invaded European coasts. Using seven microsatellites, we found virtually no genetic differentiation between natural populations throughout the European range (from the south of the Wadden Sea (the Netherlands) to the south of France) and French cultivated oysters. The genetic homogeneity of Pacific oyster samples appears to be the result of repeated transfers from same seed stocks made for aquaculture and, to a lesser extent, widespread dispersal due to specific biological traits of this species. The only genetic differentiation of Sylt population in the north of the Wadden Sea (Germany) suggests a stronger, persistent impact of ongoing supply of new genetic material from hatchery production, corresponding to seeds selection made by breeders. All of our genetic data highlighted the importance of aquaculture practices on the genetic structure of the keystone invader C. gigas in Europe.     

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.     

Sympatric size variants of the microcopepod Oncaea venusta exhibit distinct lineages in DNA sequences

We investigated the phylogenetic relationships among different size groups of the pelagic microcopepod Oncaea venusta Philippi, 1843, by comparing the patterns of genetic variation of specimens collected at five locations of the Indo-West Pacific Ocean. Phylogenetic analyses were based on sequence data obtained from two DNA markers: A 310 bp fragment of the mitochondrial cytochrome b (cyt b) gene and a 480 bp fragment of the nuclear internal transcribed spacer 1 (ITS1). The cyt b sequences showed a much higher level of variation than those from ITS1, but the conclusion from both genes was concordant. Four genetic clades could be differentiated. A small- and a large-size group were unambiguously assigned to two distinct clades or lineages. Unexpectedly, the medium-sized individuals could be divided into another two different genetic clades. All four lineages were supported by high bootstrap values. The high levels of sequence divergence under sympatric conditions indicated that at least the two main groups, the large and the small one, may be assigned to different species. For the medium-size group additional morphological studies and more sensitive nuclear markers are required to clarify their taxonomic status.     

Repeated trans-Arctic invasions in littoral bivalves: molecular zoogeography of the<Emphasis Type="Italic"> Macoma balthica</Emphasis> complex

From a geographical survey of allozyme variation, a history of repeated trans-Arctic invasions since the Plio-Pleistocene is suggested for circumboreal bivalves of the Macoma balthica complex. A principal genetic subdivision, involving several nearly diagnostic loci and Nei's distance D=0.6, distinguishes the clams of the NE Pacific from those of the NE Atlantic. The Pacific taxon is however also present in Europe, in disjunct isolates in the Baltic Sea and White Sea basins. Nevertheless, these populations have marked Atlantic introgressive elements in their gene pools (ca. 30%). Two further population types are recognized, one in the St. Lawrence estuary, Quebec, the other in Varangerfjorden, NE Norway; the latter appears a mixture of Pacific and Atlantic components in almost equal proportions, in local genetic equilibrium (a hybrid swarm). Populations in temperate North America fall outside the circumboreal M. balthica complex discussed here (D=1.0), and are referred to M. petalum. In a scenario of the history and evolution of the M. balthica complex and the similarly subdivided Mytilus edulis complex, the divergence between Pacific and Atlantic taxa started after an initial introduction of Pacific ancestors to the Atlantic basin, enabled by the Pliocene opening of the Bering Strait. During the Pleistocene and Holocene, the ocean basins were, for the most part, effectively isolated, but occasional re-invasions have taken place, causing secondary contacts of the diverged bivalve types on the Atlantic coasts. The recently re-invaded Pacific taxa in northern Europe now seem to thrive only in the extreme marginal environments. Exact dating of the re-invasions is not possible from current data. Apart from the divergence through isolation, hybridization and introgression have significantly molded the present affinities within the M. balthica complex. A formal taxonomic treatment of reticulate and hybridizing lineages is problematic; yet to recognize the evolutionary and systematic diversity within the M. balthica complex, a subspecies distinction between the NE Atlantic clams and those from the Pacific, Baltic and White Sea basins is suggested.Communicated by L. Hagerman, Helsingør     

Mitochondrial DNA sequence variation and phylogeography of oceanic insects (Hemiptera: Gerridae: Halobates spp.)

Relatively few insects have invaded the marine environment, and only five species of sea skaters, Halobates Eschscholtz (Hemiptera: Gerridae), have successfully colonized the surface of the open ocean. All five species occur in the Pacific Ocean, H. germanus White also occurs in the Indian Ocean, whereas H. micans Esch- scholtz is the only species found in the Atlantic Ocean. We sequenced a 780 bp long region of the mitochondrial cytochrome oxidase subunit I gene (COI) for a total of 66 specimens of the five oceanic Halobates species. Our purpose was to investigate the genetic variation within species and estimate the amount of gene flow between populations. We defined 27 haplotypes for H. micans and found that haplotype lineages from each of the major oceans occupied by this species are significantly different, having sequences containing five to seven unique base substitutions. We conclude that gene flow between populations of H. micans inhabiting the Atlantic, Pacific, and Indian Ocean is limited and hypothesize that these populations have been separated for 1 to 3 million years. Similarly, there may be limited gene flow between H. germanus populations found in the Pacific and Indian Ocean and between H. sericeus populations inhabiting the northern and southern parts of the Pacific Ocean. Finally, we discuss our findings in relation to recent hypotheses about the influence of oceanic diffusion on the distribution and population structure of oceanic Halobates spp. Received: 29 July 1999 / Accepted: 23 November 1999     

Isolation and population divergence of a widespread Indo-West Pacific marine gastropod at Easter Island

Oceanic islands represent excellent systems for studying the link between geographic isolation and population divergence. Easter Island is the world’s most isolated island and exhibits a high level of endemicity in the nearshore marine environment. Yet few studies have examined the effect of such extreme isolation on the divergence of populations of widespread species that occur at Easter Island. Conus miliaris, a marine gastropod distributed throughout much of the Indo-West Pacific, occurs at Easter Island where the population is ecologically and morphologically distinct from other populations of the species. To determine whether these phenotypic differences are associated with genetic isolation of the Easter Island population, we investigated the phylogeography of this species by examining mitochondrial COI sequences obtained from 141 individuals from eight localities occurring predominantly in the western, central and southeastern Pacific. Results from our analyses show that C. miliaris at Easter Island differs genetically from other populations. We estimate that C. miliaris colonized Easter Island shortly after the origin of the island ≤0.7 million years ago and that since population founding, gene flow has occurred predominantly from Easter Island to the west and that little migration has occurred into Easter Island.     

Distribution of coccolithophores in marginal seas along the western Pacific Ocean and in the Red Sea

The distribution of coccolithophores was studied in the neritic environment along the western margin of the Pacific Ocean: the Inland Sea of Seto, Yellow Sea, East China Sea, South China Sea, Java Sea, Timor Sea, Arafura Sea and Gulf of Carpentaria. The coccolithophore community in the Red Sea was also studied for comparison with the Pacific marginal seas. With minor exceptions, the coccolithophore communities were very similar throughout the neritic areas investigated, but differed completely from the pelagic community in three aspects. Firstly, almost all neritic coccolithophores, regardless of species, suffered various degrees and forms of malformation with relation to the morphology of their coccoliths, while such malformation was rare in the pelagic population. Nitrogen deficiency may cause such malformation. Secondly, the diversity of species in these marginal seas was much lower than in the pelagic environment, although no species was found to be exclusively neritic. Emiliania huxleyi, usually ubiquitous in oceanic areas and in various neritic environments of higher latitudes, was scarce, while Gephyrocapsa oceanica dominated the flora throughout the studied areas. Finally, the horizontal and vertical distributions of the neritic populations were sporadic compared to those of the rather uniform pelagic environments.     

Depth-related adaptations, speciation processes and evolution of color in the genus Phyllidiopsis (Mollusca: Nudibranchia)

The nudibranch genus Phyllidiopsis (Phyllidiidae) contains 30 currently recognized species, all of them distributed throughout the tropical Indo-Pacific, eastern Pacific, Northwest Atlantic and Caribbean Sea. Half of the known species of Phyllidiopsis inhabit deep waters, and most of the deep-sea species of the Phyllidiidae belong to this genus. There is no definitive explanation for the high diversity of Phyllidiopsis in the deep-sea or for whether diversity could be related to particular adaptations of this group or to historical events. In light of phylogenetic analysis, several cases of vicariance have been detected in this genus. Apparently two major vicariant events occurred between the tropical Indo-Pacific region and the Atlantic-eastern Pacific area first and subsequently between the eastern Pacific and the Atlantic. Vicariant events could also be involved in producing vertical distributional patterns in a few species of Phyllidiopsis. The scarcity of phyllidiids in the Atlantic Ocean may be explained by historical events, including isolation and subsequent extinction in shallow waters. There is a mimicry species complex in Phyllidiopsis, including several members of a clade that probably acquired this coloration through common ancestry, and also including another unrelated species that probably acquired this coloration through convergent or parallel evolution. There is also a group of white species, lacking any other contrasting colors, that inhabits deep waters. This coloration could constitute an adaptation to the deep-sea environment and not a mimicry complex. In this case, all species acquired this coloration through common ancestry.     

Genetic heterogeneity among <Emphasis Type="Italic">Eurytemora affinis</Emphasis> populations in Western Europe

Evolutionary diversification of the broadly distributed copepod sibling species complex Eurytemora affinis has been documented in the northern hemisphere. However, the fine scale geographic distribution, levels of genetic subdivision, evolutionary, and demographic histories of European populations have been less explored. To gain information on genetic subdivision and to evaluate heterogeneity among European populations, we analyzed samples from 8 locations from 58° to 45°N and 0° to 23°E, using 549 base pairs of the mitochondrial cytochrome oxidase subunit I (COI) gene. We discovered three distinct lineages of E. affinis in Western Europe, namely the East Atlantic lineage, the North Sea/English Channel (NSEC) lineage, and the Baltic lineage. These geographically separated lineages showed sequences divergence of 1.7–2.1%, dating back 1.9 million years (CI: 0.9–3.0 My) with no indication of isolation by distance. Genetic divergence in Europe was much lower than among North American lineages. Interestingly, genetic structure varied distinctively among the three lineages: the East Atlantic lineage was divided between the Gironde and the Loire populations, the NSEC lineage comprised one single population unit spanning the Seine, Scheldt and Elbe rivers and the third lineage was restricted to the Baltic Proper (Sweden). We revealed high haplotype diversity in the East Atlantic and the Baltic lineages, whereas in the NSEC lineage haplotype diversity was comparatively low. All three lineages showed signs of at least one demographic expansion event during Pleistocene glaciations that marked their genetic structure. These results provide a preliminary overview of the genetic structure of E. affinis in Europe.     

Incipient speciation within a subgenus of rockfish (<Emphasis Type="Italic">Sebastosomus</Emphasis>) provides evidence of recent radiations within an ancient species flock

The high frequency of speciation events associated with species flocks (i.e., radiations of closely related species) provides invaluable insight into the speciation process. Investigations of the speciation process in the marine environment are rare, and therefore, the genetic analysis of the rockfish genus Sebastes, considered an ancient marine species flock, provides an opportunity to investigate this process in the sea. Using both mitochondrial and nuclear markers, we analyzed five closely related species within the rockfish subgenus Sebastosomus. Our goal was to understand the evolutionary history and genetic relationships among species within this group and to provide evidence of recent speciation events within the subgenus. In the genetic analysis of the subgenus, we found different stages of the speciation process, with greater genetic divergences among three of the five species, evidence of recent divergence between two of the five species, Sebastes entomelas and S. mystinus, and significant genetic divergence between two lineages within S. mystinus revealing a signature of incipient speciation. We also found frequency differences of the two S. mystinus lineages among sample locations and found no evidence of introgression between the lineages at the location where both coexist. Although Sebastes is an example of an ancient species flock, this study provides evidence of ongoing speciation within the genus and reveals stages of this process from incipient to distinct species.     

Evidence of different reproductive traits in the transisthmian sister species, <Emphasis Type="Italic">Alpheus saxidomus</Emphasis> and <Emphasis Type="Italic">A. simus</Emphasis> (Decapoda,Caridea, Alpheidae): description of the first postembryonic stage

The emergence of the Isthmus of Panama, approximately 3.0-3.5 million years ago, established two very different marine systems (the Caribbean and the eastern Pacific) and separated previously continuous marine populations. The geographic isolation of transisthmian sister species provides an excellent basis for the study of divergence and speciation processes. Here we describe the morphology of the first postembryonic stage of Alpheus saxidomus and A. simus, two rock-boring alpheids; the estimated time since divergence for this transisthmian pair ranges from 4.4-6.1 to 3-9 million years. The presence of a first zoeal stage in A. saxidomus, e.g., without pleopods and a telson with 7+7 setae, indicates a prolonged developmental sequence. In contrast, hatchlings of A. simus are substantially more developed and resemble juveniles. Thus, the developmental modes of A. saxidomus and A. simus are strikingly different with abbreviated, most probably direct, development in the latter species. Reduced food availability in the Caribbean compared to the Pacific coast is likely to be a possible reason for the evolution of such important differences in life history traits of the two transisthmian sister species. It is suggested that a transition from prolonged to abbreviated development evolved gradually during the estimated time since divergence; however, such a shift may have taken place within a substantially smaller time span.     

Geographical differences in early growth of walleye pollock Theragra chalcogramma,estimated by back-calculation of otolith daily growth increments

The relationship between otolith length and body length (total length) was examined in walleye pollock Theragra chalcogramma (Pallas), collected from Pacific Ocean waters adjacent to Hokkaido, Japan, and in larvae reared from fertilized eggs. A linear relationship was found between log-transformed body length and otolith length data with two inflection points at ca. 11 mm and 100 mm total length. This relationship was found to be applicable also to samples from the Japan Sea, Bering Sea and Okhotsk Sea. The early growth pattern estimated by back-calculation of otolith increments of 1-yr fish from Funka (Uchiura) Bay (Hokkaido) accorded with that obtained from size-at-age data of 0 yr fish collected in the preceding year. Differences were found in comparisons of the back-calculated early growth pattern between samples from waters adjacent to Hokkaido, the Pacific Ocean, the Japan Sea and the Okhotsk Sea. The growth curve of the Okhotsk samples was markedly different from others, showing rapid initial growth up to about 100 d after hatching and attaining a small body size in the first year.     

Population structure of the planktonic copepod Calanus pacificus in the North Pacific Ocean

The pelagic copepod Calanus pacificus ranges nearly continuously across temperate-boreal regions of the North Pacific Ocean and is currently divided into three subspecies—C. pacificus oceanicus, C. pacificus californicus, C. pacificus pacificus—based on subtle morphological differences and geographic location. The relation between geography and genetic differentiation was examined for 398 C. pacificus individuals sampled from six widely distributed locations across the North Pacific, including an open ocean site and coastal sites on both sides of the North Pacific basin. For each individual copepod, the DNA sequence was determined for a 421-bp region of the mitochondrial coxI gene (mtCOI). A total of sixty-three different mtCOI sequences, or haplotypes, were detected, with a sequence divergence between haplotypes of 0.2–3.1%. The number and distribution of haplotypes varied with sampling location; 12 haplotypes were distributed across multiple sampling locations, and 51 occurred at only one location. Five genetically distinct populations were detected based on F _ST values. Haplotype minimum spanning networks, nucleotide divergence and F _ST values indicated that individuals from coastal sites in the North Pacific Ocean were more closely related to each other than to individuals from the open ocean site at Station P. These results provide genetic support for the designation of two subspecies—a coastal subspecies that consists of what is currently referred to as C. p. pacificus and C. p. californicus and an open ocean subspecies C. p. oceanicus. This work also indicates that planktonic copepods with potentially high dispersal capacity can develop genetically structured populations in the absence of obvious geographic barriers between proximate locales within an ocean basin.