Variability of DNA microsatellite loci in populations of Pacific cod <Emphasis Type="Italic">Gadus macrocephalus</Emphasis> Tilesius (Gadidae)

Despite almost a hundred years of studies on the Pacific cod’s biological and ecological peculiarities, many of them and especially population structure remain poorly understood. The variability of DNA microsatellite loci Gmo3, Gmo34, Gmo35, Pgmo32, and Gmo 19 was analyzed for the cod sampled in different areas of the North Pacific. The cod sampled nearby the Southern Kurils, differed significantly by Gmo3 and Pgmo32 compared to the populations inhabiting the Bering Sea, the Sea of Okhotsk and the coastal waters off Canada. Further, Pacific cod of the three latter populations demonstrated high similarity (I = 0.996) in spite of considerable geographic remoteness of these areas from one another.     

On genetic differentiation of the Pacific cod <Emphasis Type="Italic">Gadus macrocephalus</Emphasis> tilesius, 1810 (Gadiformes: Gadidae)

The variability of the Gmo3, Gmo34, Gmo35, and Pgmo32 DNA microsatellite loci in Pacific cod Gadus macrocephalus samples from different areas of the North Pacific was analyzed. The data obtained show that Pacific cod from the southern Kuril Islands significantly differs from the populations of the Bering Sea, the Sea of Okhotsk, and the coastal waters of Canada (the microsatellite loci Gmo3 and Pgmo32 bear the highest differentiating capacity). Despite the significant geographical remoteness of these areas from one another, the above three Pacific cod populations exhibit a high degree of similarity (I = 0.997–0.999).     

Preliminary data on the variability of three microsatellite loci in Pacific <Emphasis Type="Italic">Gadus macrocephalus</Emphasis> and Atlantic <Emphasis Type="Italic">G. morhua</Emphasis> cod (Gadidae)

Variability of microsatellite DNA loci Gmo3, Gmo34, and Gmo35 is studied in samples of Pacific cod Gadus macrocephalus and Atlantic cod G. morhua. The results show high values of identity of the samples within the North Pacific basin (0.9766–0.9924) and within the Northeast Atlantic basin (0.9580). Based on the pairwise assessment of genetic differentiation, the F _ST values are significantly different in all variants between the samples of Pacific and Atlantic cod (F _ST = 0.5235–0.6719, p < 0.001). Within the basins, the significant differences in the frequencies of main alleles are revealed in the loci Gmo3 and Gmo34 for the samples from the Pacific and Atlantic oceans, respectively.     

Variability of microsatellite loci of Greenland cod <Emphasis Type="Italic">Gadus ogac</Emphasis> Richardson 1836: Comparison with other species of <Emphasis Type="Italic">Gadus</Emphasis> genus (Gadidae)

Comparative analysis of variability of seven microsatellite loci—Gmo3, Gmo-G12, Gmo-G18, Gmo19, Gmo34, Gmo35 and Pgmo32—was performed for the Greenland cod Gadus ogac, Pacific cod G. macrocephalus, Atlantic cod G. morhua, and White Sea cod G. morhua marisalbi. High genetic identity was observed between the Greenland cod and Pacific cod (I = 0.9520). Pair analysis of genetic differentiation was performed on the studied microsatellite loci according to θ (analogue of F _ST). The Greenland cod differed significantly from the Pacific, Atlantic, and the White Sea cod; however, the differentiation level varied. The lowest value was observed for the pair Greenland cod-Pacific cod (0.123), and the highest levels were registered for the pairs Greenland cod-Atlantic cod (0.605) and Greenland cod-White Sea cod (0.535).     

Evidence of microsatellite hitch-hiking selection in Atlantic cod (Gadus morhua L.): implications for inferring population structure in nonmodel organisms

Microsatellites have gained wide application for elucidating population structure in nonmodel organisms. Since they are generally noncoding, neutrality is assumed but rarely tested. In Atlantic cod (Gadus morhua L.), microsatellite studies have revealed highly heterogeneous estimates of genetic differentiation among loci. In particular one locus, Gmo 132, has demonstrated elevated genetic differentiation. We investigated possible hitch-hiking selection at this and other microsatellite loci in Atlantic cod. We employed 11 loci for analysing samples from the Baltic Sea, North Sea, Barents Sea and Newfoundland covering a large part of the species' distributional range. The 'classical' Lewontin-Krakauer test for selection based on variance in estimates of F(ST) and (standardized genetic differentiation) revealed only one significant pairwise test (North Sea-Barents Sea), and the source of the elevated variance could not be ascribed exclusively to Gmo 132. In contrast, different variants of the recently developed ln Rtheta test for selective sweeps at microsatellite loci revealed a high number of significant outcomes of pair-wise tests for Gmo 132. Further, the presence of selection was indicated in at least one other locus. The results suggest that many previous estimates of genetic differentiation in cod based on microsatellites are inflated, and in some cases relationships among populations are obscured by one or more loci being the subject to hitch-hiking selection. Likewise, temporal estimates of effective population sizes in Atlantic cod may be flawed. We recommend, generally, to use a higher number of microsatellite loci to elucidate population structure in marine fishes and other nonmodel species to allow for identification of outlier loci that are subject to selection.     

New Nuclear SNP Markers Unravel the Genetic Structure and Effective Population Size of Albacore Tuna (Thunnus alalunga)

In the present study we have investigated the population genetic structure of albacore (Thunnus alalunga, Bonnaterre 1788) and assessed the loss of genetic diversity, likely due to overfishing, of albacore population in the North Atlantic Ocean. For this purpose, 1,331 individuals from 26 worldwide locations were analyzed by genotyping 75 novel nuclear SNPs. Our results indicated the existence of four genetically homogeneous populations delimited within the Mediterranean Sea, the Atlantic Ocean, the Indian Ocean and the Pacific Ocean. Current definition of stocks allows the sustainable management of albacore since no stock includes more than one genetic entity. In addition, short- and long-term effective population sizes were estimated for the North Atlantic Ocean albacore population, and results showed no historical decline for this population. Therefore, the genetic diversity and, consequently, the adaptive potential of this population have not been significantly affected by overfishing.     

Data on variation of microsatellite loci in Kildin cod <Emphasis Type="Italic">Gadus morhua kildinensis</Emphasis> (Gadidae)

Variation of microsatellite loci Gmo8, Gmo-G12, Gmo-G18, Gmo19, PGmo32, Gmo34, and Gmo35 is investigated in Kildin cod Gadus morhua kildinensis. The investigated loci are characterized by a low level of variation: five loci are represented by two-allele systems and three loci are monomorphic. Mean value of heterozygosity calculated by all investigated loci is lower in Kildin cod than in Atlantic Gadus morhua—0.2854 vs. 0.5667.     

EVOLUTION OF ATLANTIC AND PACIFIC COD: LOSS OF GENETIC VARIATION AND GENE EXPRESSION IN PACIFIC COD

An allozyme investigation of 41 protein-coding loci in two morphologically similar fishes, Atlantic and Pacific cod, indicates that Pacific cod experienced a severe population bottleneck that led to the loss of gene diversity and gene expression. Pacific cod possesses a significantly lesser amount of gene diversity (H = 0.032) than Atlantic cod (H = 0.125) and lacks gene expression for Me-3. Allele-frequency distributions differ between species as predicted by neutral theory: Atlantic cod has a U-shaped distribution, which is expected for populations in drift-mutation equilibrium, whereas Pacific cod has a J-shaped distribution with an excess of low-frequency alleles. This excess may be explained by the appearance of new alleles through mutation which have not yet reached intermediate frequencies through drift. The population bottleneck in Pacific cod was most likely associated with founder populations that dispersed into the Pacific Ocean after the Bering Strait opened. Under the molecular-clock hypothesis a Nei genetic distance of 0.415 (based on 41 loci) suggests that Pacific cod dispersed into the Pacific Ocean soon after the Bering Strait opened in the mid-Pliocene, 3.0 to 3.5 million years ago.     

Characteristic features of ecology of the Pacific saury Cololabis saira (Scomberesocidae, Beloniformes) in open waters and in the northeast Pacific ocean

The results of long-term studies of the spatial distribution, size composition, and spawning intensity of the Pacific saury Cololabis saira in open waters of the North Pacific Ocean and the northeastern part of the Pacific Ocean off the west coast of North America are presented. In the open waters of the Pacific Ocean, saury forms aggregations early in summer; intraseasonal changes in their distribution pattern have been identified. Migrations of Pacific saury stocks in the northeastern part of the Pacific Ocean were considered, and assumptions about their causes were made. The differences in the size composition of Pacific saury in different parts of the range are associated with the peculiarities of occurrence of favorable conditions for the formation of aggregations. Intensive spawning of Pacific saury in the northeastern part of the Pacific Ocean is observed in winter and spring, and that in open waters is observed in summer. The distribution of the early progeny of Pacific saury is associated with the systems of currents; its findings in the extreme northern regions in the 1980s are explained.     

Population Structure of Pacific Cod <Emphasis Type="Italic">Gadus macrocephalus</Emphasis> in the Southern Part of the Range Based on the Microsatellite Analyses

The population structure of Pacific cod Gadus macrocephalus in the southern part of the range and adjacent regions is studied on the basis of the results of microsatellite analyses. Collected data indicate heterogeneity of this species population within the studied area. According to the obtained F_ST values, Pacific cod from the waters of the Republic of Korea (Yellow Sea side) and northwestern part of the Sea of Okhotsk significantly differ from all other studied regions (Table 4). Significant differentiation was also revealed between samples from the waters of the Tatar Strait and all other regions except for South Kurils Pacific cod (both Sea of Okhotsk and Pacific Ocean sides). These two latter sample collections were similar to each other as well. A low level of differentiation was also shown for the Peter the Great Bay and the East Sea/Sea of Japan waters of the Republic of Korea.     

Isolation and Characterization of Tetranucleotide Microsatellites from Atlantic Haddock (Melanogrammus aeglefinus)

Five tetranucleotide microsatellite loci developed from Atlantic haddock (Melanogrammus aeglefinus) are presented. Loci were isolated using a modified magnetic bead-hybridization selection procedure that enriched for tetranucleotide microsatellites. Loci were polymorphic (3–99 alleles per locus; mean, 33.2) and exhibited high levels of observed heterozygosity (0.07–0.99; mean, 0.73) in a sample of 70 haddock collected from the Scotian Shelf in the Northwest Atlantic Ocean. Primer sets for 5 tetranucleotide microsatellites, originally developed from cod (Gmo34), were also tested in Atlantic haddock; one pair yielded readily detectable product and was variable in the population assayed (29 alleles; heterozygosity, 0.96). These loci are suitable for kinship analyses in aquaculture-related applications, and are potentially useful for resolving population structure in the wild.     

Stable Isotopic and Trace Elemental Compositions of Otoliths and the Stock Structure of Pacific Cod, Gadus macrocephalus

Synopsis We examined the stock composition and life history of Pacific cod, Gadus macrocephalus, in the northeast Pacific Ocean from sagittal otoliths collected from three marine fishing areas in Washington State, U.S.A. We analyzed both stable isotope ratios (δ¹⁸O and δ¹³C) and trace elemental concentrations (Sr, Mg, Na, Fe, Mn) for these otoliths. The combination of δ¹⁸O and δ¹³C, and correlation of δ¹⁸O vs. 1000Sr/Ca and 1000Mg/Ca showed clear separations between North Puget Sound and coastal cod, suggesting there might be two different spawning stocks in the region. The North Puget Sound cod might represent an ‘Estuary-type’ from the Strait of Georgia, whereas coastal cod might represent an ‘Ocean-type’ from the Pacific west coast. Isotopic variations from five representative otoliths also showed a two-stage life history for Pacific cod, and a critical transition period for cod in migration to the ocean or in age of sexual maturity. These chemical interpretations and conclusions appear in agreement with biological observations of Pacific cod, and are consistent with results of previous studies for other marine fish species in nearby areas.     

Multiple population structure of the giant mottled eel, Anguilla marmorata

The population structure of the giant mottled eel, Anguilla marmorata, was investigated with mitochondrial and microsatellite DNA analyses using 449 specimens from 13 localities throughout the species range. Control region F-statistics indicated the North Pacific (Japan, Taiwan, Philippines, Sulawesi), South Pacific (Tahiti, Fiji, New Caledonia, Papua New Guinea), eastern Indian Ocean (Sumatra), western Indian Ocean (Réunion, Madagascar), Ambon, and Guam regions were significantly different (Phi(ST) = 0.131-0.698, P < 0.05) while only a few differences were observed between localities within the South Pacific. These regions were roughly clustered in the neighbour-joining tree, although Ambon individuals were mainly divided into North and South Pacific groups. Analysis with eight microsatellite loci showed almost identical results to those of the control region, except no genetic difference was observed between the western and eastern Indian Ocean (F(ST) = 0.009, P > 0.05). The Bayesian cluster analysis of the microsatellite data detected two genetic groups. One included four North Pacific localities, and the other included eight localities in the South Pacific, Indian Ocean, and Guam, but Ambon individuals were evenly assigned to these two groups. These results showed that A. marmorata has four genetically different populations (North Pacific, South Pacific, Indian Ocean, Guam region). The North Pacific population is fully panmictic whereas the South Pacific and Indian Ocean populations have a metapopulation structure. Interestingly, Guam was suggested to be inhabited by a reproductive population restricted to that region, and the individuals from the North and South Pacific populations co-exist in Ambon.     

Diversity of the <Emphasis Type="Italic">CO1</Emphasis> Gene of Mitochondrial DNA in Representatives of Genus <Emphasis Type="Italic">Antimora</Emphasis> (Moridae,Gadiformes) in the World Oceans

The frequency of occurrence of the COI gene of mitochondrial DNA in the Pacific flatnose Antimora microlepis and blue antimora Antimora rostrata (Moridae, Gadiformes) was analyzed in samples collected in different areas of the World Ocean. The revealed maximum haplotype diversity of COI in the blue antimora in the North Atlantics may indicate that this species emerged in this region, from which it widely distributed in the World Ocean. The Pacific flatnose Antimora microlepis originated from Antimora rostrata. Antimora might penetrate into the North Pacific by several routes: through the Panama Strait, along the coast of Antarctica, or through the Indian Ocean along the coast of Australia.     

Simultaneous Analysis of Six Microsatellite Markers in Atlantic Cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>): A Novel Multiplex Assay System for Use in Selective Breeding Studies

A novel hexaplex assay system including Gmo8, Gmo19, Gmo35, Gmo37, Tch11, and Tch12 microsatellites from Atlantic cod, consisting of trinucleotide or tetranucleotide repeat units, is introduced. All 6 loci were coamplified in a single reaction employing dye-labeled primers. Alleles from these loci were sized using an internal standard by automated sample processing in an ABI 310 Genetic Analyser. Amplified alleles in profiles containing selected microsatellites were typed clearly, providing easily interpretable results. Sequencing data indicated that alleles at all loci consisted of simple repeat units. This may help minimize the likelihood of stuttering upon polymerase chain reaction amplification. The results suggest that the presented hexaplex assay system may be a useful tool in a selective breeding program in which genetic identification will allow different genotypes to be reared together from fertilization. This should have a great impact as it will make selective breeding more efficient.     

Characteristics of under-ice swarming of polar cod Boreogadus saida (Gadidae) in the Central Arctic Ocean

Phenological observations on under-ice swarming behavior of Polar cod Boreogadus saida were performed during the winter period 2009–2010 in the Canadian sector of the Arctic Ocean on the North Pole NP-37 drifting station. These observations and ichthyologic collections were the basis of the present study. The schools were mostly formed by the immature specimens (TL = 75–169 mm; 1+ to 4+ age), the dominating group was presented by fish of 2+ age (86–94% of total abundance). The driving factors of such large under-ice swarming of Polar cod B. saida in the Arctic are discussed, particularly, the interactions of Polar cod development and the ice drift in the studied region. This species inhabits the overcooled water (?1.8°C) under the ice, so the scheme of the antifreeze agent producing in the fish blood is considered. We make assumptions about under ice migrations of Polar cod swarming.     

Genetic Diversity and Population Structure of the Pelagic Thresher Shark (Alopias pelagicus) in the Pacific Ocean: Evidence for Two Evolutionarily Significant Units

There has been an increasing concern about shark overexploitation in the last decade, especially for open ocean shark species, where there is a paucity of data about their life histories and population dynamics. Little is known regarding the population structure of the pelagic thresher shark, Alopias pelagicus. Though an earlier study using mtDNA control region data, showed evidence for differences between eastern and western Pacific populations, the study was hampered by low sample size and sparse geographic coverage, particularly a lack of samples from the central Pacific. Here, we present the population structure of Alopias pelagicus analyzing 351 samples from six different locations across the Pacific Ocean. Using data from mitochondrial DNA COI sequences and seven microsatellite loci we found evidence of strong population differentiation between western and eastern Pacific populations and evidence for reciprocally monophyly for organelle haplotypes and significant divergence of allele frequencies at nuclear loci, suggesting the existence of two Evolutionarily Significant Units (ESU) in the Pacific Ocean. Interestingly, the population in Hawaii appears to be composed of both ESUs in what seems to be clear sympatry with reproductive isolation. These results may indicate the existence of a new cryptic species in the Pacific Ocean. The presence of these distinct ESUs highlights the need for revised management plans for this highly exploited shark throughout its range.     

Development and characterization of novel di‐ and tetranucleotide microsatellite markers in Pacific cod (Gadus macrocephalus)

The Pacific cod (Gadus macrocephalus) supports large commercial fisheries in the northern Pacific Ocean and the Bering Sea. Here we characterize 10 polymorphic microsatellite loci isolated from enriched genomic libraries. Loci were screened on a sample of 96 spawning adults. The average number of alleles per locus was 25.3 (range 12–44), with expected heterozygosities (H_E) ranging from 0.54 to 0.97. No significant deviations of genotypic proportions from Hardy–Weinberg equilibrium or linkage equilibrium were detected. These markers will be used in future studies of population structure and mixed stock analysis of this important gadid species.     

Temperature-dependent growth and behavior of juvenile Arctic cod (<Emphasis Type="Italic">Boreogadus saida</Emphasis>) and co-occurring North Pacific gadids

The thermal sensitivity of Arctic fish species is poorly understood, yet such data are a critical component of forecasting and understanding ecosystem impacts of climate change. In this study, we experimentally measured temperature-dependent growth and routine swim activity in the juvenile stage of two Arctic gadids (Arctic cod, Boreogadus saida and saffron cod, Eleginus gracilis) and two North Pacific gadids (walleye pollock, Gadus chalcogrammus and Pacific cod, Gadus macrocephalus) over a 6-week growth period across five temperatures (0, 5, 9, 16 and 20 °C). Arctic cod demonstrated a cold-water, stenothermic response in that there was relatively high growth at 0 °C (0.73 % day^?1), near-maximal growth at 5 °C (1.35 % day^?1) and negative impacts on activity, growth and survival at 16 °C. In contrast, saffron cod demonstrated a warmer-water, eurythermic response, and temperature had a positive effect on growth and condition beyond 16 °C. However, despite these distinct thermal responses, walleye pollock and Pacific cod grew 2–3 times faster than Arctic gadids across a relatively broad temperature range above 5 °C. These results, coupled with possible northward expansion by both Pacific cod and walleye pollock, suggest Arctic cod are highly vulnerable to continued climate change in the Arctic, especially in coastal areas of the Beaufort and Chukchi Seas where temperatures already exceed 14 °C in the summer growth period.     

北太平洋柔鱼微卫星标记的筛选及遗传多样性

采用(AC)12、(AG)12两种生物素探针,通过磁珠富集法构建了柔鱼部分基因组微卫星富集文库。68个阳性克隆中有60个含有微卫星序列,重复次数在10次以上的占86.84%,最高重复次数为33次。其中,完美型微卫星占60.53%,非完美型微卫星占36.84%,混合型微卫星占2.63%。除探针使用的AC/TG、AG/TC重复外,还得到ACAG、AGAC重复序列。利用筛选出的8个微卫星位点对北太平洋柔鱼6个群体的遗传多样性及遗传结构进行分析。结果表明,8个微卫星位点均为高度多态性位点(PIC=0.787—0.987),位点Bo103与位点Bo105极显著偏离Hardy-Weinberg平衡(P0.01)。6个地理位置的柔鱼群体显示出较高的遗传多样性水平(Ho=0.672—0.761,He=0.808—0.851)。两两群体间的Fst值以及AMOVA分析结果均表明,群体间遗传分化不显著(Fst=0.00559,P0.05),遗传差异主要来自于个体间。基于Nei's遗传距离的UPGMA聚类树显示,北太平洋东北部2个柔鱼群体(NE1、NE3)聚为一类,西北部3个群体(NW1、NW2、NW3)与东北部1个群体(NE2)另聚为一类,且群体NW1与群体NE2亲缘关系最近,遗传距离与地理距离线性相关分析没有呈现出正相关性(R=0.175,P0.05)。遗传结构分析结果推断北太平洋柔鱼存在1个理论群。柔鱼个体具有较强的游泳能力,在海流的作用下,群体之间存在较强的基因交流。建议今后在柔鱼资源开发利用过程中将北太平洋柔鱼看作1个管理单元。