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).     

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 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.     

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.     

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.     

Formation of genetic diversity in populations of pacific cod (Gadus macrocephalus Tilesius) (Gadidae)

The variability of the Gmo3, Gmo34, Gmo35, Pgmo32, Gmo19 microsatellite DNA loci was examined in the population samples of Pacific cod from different parts of the North Pacific Ocean. The statistical tests showed that the differentiation of the population was the result of different levels of reproductive autonomy of the individual Pacific cod populations, formed in the Pleistocene-Holocene period with the specific features of transgressions and regressions of the World Ocean, and supported by the existing system of currents and the features of the migratory behavior of cod in North Pacific.     

New Data on the Biology of Kildin Cod Gadus morhua kildinensis (Gadidae) from Lake Mogilnoe (Kildin Island,the Barents Sea) Obtained by Echo Sounding and Underwater Photography

Journal of Ichthyology - The distribution of the Kildin cod Gadus morhua kildinensis in Lake Mogilnoye (Kildin Island, Barents Sea) is assessed based on the results of echo sounding and...     

Polymorphic microsatellite loci for species of Australian freshwater cod, <Emphasis Type="Italic">Maccullochella</Emphasis>

The Australian freshwater cod genus, Maccullochella is represented by three species: Murray cod, M. peelii peelii, eastern freshwater cod, M. ikei, and trout cod, M.macquariensis. Seven novel microsatellite loci from M. ikei and six previously published loci from M. peelii peelii were tested on wild populations of Murray, eastern and trout cod. Levels of polymorphism varied between species with 13 loci polymorphic in Murray cod, 9 in trout cod and 7 in eastern cod. Observed heterozygosities ranged from 0.053 to 0.842. This suite of microsatellite loci will facilitate future studies of the genetic status of wild and hatchery bred populations of Maccullochella.     

Mechanisms of the adaptation of the Kildin cod <Emphasis Type="Italic">Gadus morhua kildinensis</Emphasis> Derjugin, 1920 (Pisces: Gadidae) to the specific conditions of Lake Mogilnoye

The mechanisms of the adaptation of the Kildin cod to the conditions of a meromictic lake were analyzed based on the study of morphological, biological, and genetic characteristics. Significant differences between the Kildin cod and Atlantic cod have been found in their morphological and genetic characteristics. The results of an X-ray microanalysis of otoliths, which were conducted for the first time, confirm that the distribution range of the Kildin cod in Lake Mogilnoye also extends to brackish waters. It has been shown that the formation of the reproductively isolated lacustrine population of Kildin cod is accompanied by specialization in a number of characteristics (body color, feeding strategy, behavior, etc.) and by a decrease in genetic diversity.     

Identification and characterisation of nine new gene-associated microsatellite markers for Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis> L.)

Nine polymorphic microsatellite markers were identified by screening of 2464 ESTs derived from a cDNA library of Atlantic cod (Gadus morhua L.). About 35 novel microsatellite loci were selected and characterised in 96 individual cod. Nine markers were successfully amplified with number of alleles from 3 to 18 per locus and the average heterozygosity was 0.57 in the panel examined (range 0.29–0.86). All loci followed the Hardy–Weinberg expectation and no significant linkage disequilibrium was found in a test including all pairwise combinations. The gene identity was determined at four of the loci, confirming the associated microsatellites as Type I markers.     

Identification and characterisation of thirteen new microsatellites for Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis> L.) from a repeat-enriched library

A total of 13 polymorphic microsatellite markers were developed for Atlantic cod (Gadus morhua L.) from a repeat-enriched library. Polymorphism of each locus was assessed in 96 unrelated individuals from a natural population. The number of alleles per locus varied from 8 to 45. The ranges of observed and expected heterozygosity were 0.122–0.907 and 0.673–0.965, respectively. Four of the loci (Gmo-G24, Gmo-G40, Gmo-G46 and Gmo-G49) followed Hardy–Weinberg expectation. No evidence for linkage disequilibrium between pairs of loci was found in any combination of loci pairs, except between Gmo-G40 and Gmo-G43. These microsatellite markers provide useful tools for studies of population genetics, reproductive ecology and for constructing linkage maps of Atlantic cod. Jon-Ivar Westgaard and Tekle Tafese have contributed equally to the work.     

Toward <Emphasis Type="Italic">Gadus</Emphasis> (Gadidae) genus taxonomy: Development of modern structure

According to the results of analysis of genetic differentiation and Bayesian hierarchical cluster analysis of polymorphism in microsatellite loci, it was demonstrated that the development of marginal groups represented by subspecies level taxa (Greenland (Gadus macrocephalus ogac) and White Sea (Gadus morhua marisalbi) cod) occurred in the Late Holocene in a similar scenario, but in different regions and from different initial forms. The development of these reproductively independent cod groups was a result of physiological adaptations to arctic conditions in coastal northern water areas that developed under global cooling in the Subboreal.     

Changes in the diet of the coastal cod <Emphasis Type="Italic">Gadus morhua marisalbi</Emphasis> in the Kandalaksha Gulf of the White Sea under conditions of increased abundance of three-spined stickleback <Emphasis Type="Italic">Gasterosteus aculeatus</Emphasis>

The data on feeding of coastal cod Gadus morhua marisalbi in the area of the Chupa Bay (Kandalaksha Gulf of the White Sea) are presented. In the food spectrum of cod, over 25 cm long fish and their eggs dominated (77.5% by frequency of occurrence and 91.7% by weight). Other groups of food organisms occupied a secondary place. Among fish, three-spined stickleback Gasterosteus aculeatus was the most prominent food item in the diet of cod (48.3% by weight). The published data on long-term changes of the diet of cod are considered, which are mostly related to considerable fluctuations of the abundance of three-spined stickleback in the White Sea. It is shown that, at present, three-spined stickleback again plays an important role in the diet of the White Sea cod     

0-group cod(Gadus morhua) in captivity: Differential survival of certain genotypes

Genotypes atLDH-3 (lactate dehydrogenase) andPGI-1 (phosphoglucoisomerase) in 263 0-group specimens of Atlantic cod(Gadus morhua) caught in Trondheimsfjorden, Norway, in fall 1983 showed significantly different survival rates during 72 days in captivity. The heterozygote was nominally superior at both loci, and there was a significant accumulation of double heterozygotes among the survivors. Apparently,LDH-3 andPGI-1 are not selectively neutral, and allele frequency differences at these two loci between groups of cod in nature should not be interpreted as markers of reproductive isolation. The results are related to current concepts of cod population structure.Contribution No. 231, Biological Station, N-7000 Trondheim, Norway     

First evidence of spawning of eastern Baltic cod (Gadus morhua callarias) in the Belt Sea,the main spawning area of western Baltic cod (Gadus morhua L.)

Two cod stocks (western Baltic cod, WBC, and eastern Baltic cod, EBC) are managed in the Baltic Sea which is characterized by different main spawning areas and different main spawning periods. In this study we analyse the spatial and temporal occurrence of spawning individuals of both cod stocks in the main spawning grounds of the Baltic Sea based on eight microsatellite loci. Our results suggest that EBC (Gadus morhua callarias) has formed currently temporally stable, substantially homogeneous population not only in the Bornholm Sea (ICES SD: 25) but also in the Arkona Sea (ICES SD: 24). The presented analyses proved that EBC (G. m. callarias) can temporarily also spawn in the Belt Sea.     

Oxygen consumption in four species of teleosts from Greenland: no evidence of metabolic cold adaptation

Standard metabolic rate of Greenland cod or uvak, Gadus ogac, polar cod, Boreogadus saida, Atlantic cod, Gadus morhua, and sculpin, Myxocephalus scorpius, caught in the same geographical area on the west coast of Greenland was measured at 4.5°C, the temperature at which the fish were caught. The present data does not support the Metabolic Cold Adaptation theory in the traditional sense of the standard metabolic rate being 2–4 times higher for Arctic fishes than for temperate species. The standard metabolic rate of the two exclusively Arctic species of teleosts was only 10% and 26% higher, respectively, than the two species that occur in temperate as well as Arctic areas. The critical oxygen tension, with respect to oxygen consumption, of resting uvak was between 50 and 60 mmHg, and the lethal oxygen tension 20–25 mmHg at 4.5°C, which is considerably higher than for Atlantic cod from a temperate area measured at the same temperature.     

Rapid Evolution of the Intergenic T–P Spacer in the mtDNA of Arctic Cod <Emphasis Type="Italic">Arctogadus glacialis</Emphasis>

A noncoding intergenic spacer has previously been reported in mtDNA of Gadiformes. Here we present sequence information from two other cod species and variation within three species to clarify the evolution of this region. A general feature of the T–P spacer is high variation and folding into two or three hairpins. The variation among species both in structure of the region and sequence variation reflects the phylogenetic relationship of the species. A unique pattern is found within Arctic cod, Arctogadus glacialis, in which tandem repeat motifs result in new stable secondary structures. There is large variation in size of the region both within (heteroplasmy) and among individuals. A duplicated insertion is found in Greenland cod, Gadus ogac, at the same position as a corresponding duplication in Atlantic cod, Gadus morhua.     

Nine new polymorphic microsatellite loci for the amplification of archived otolith DNA of Atlantic cod,Gadus morhua L.

Nine out of 22 microsatellite primers tested were successfully amplified on three samples of cod Gadus morhua L. (two contemporary and one archived otolith samples). All loci were polymorphic (5–23 alleles/locus). The average observed heterozygosity across loci and samples was 0.625, ranging from 0.294 to 0.895 at each locus. All loci were under Hardy–Weinberg equilibrium, except PGmo56 that showed significant excess of heterozygotes in all studied samples. The isolated loci were suitable for degraded DNA and therefore useful for conducting a long‐term temporal study with DNA obtained from archived otoliths of cod.     

Mixed stock analysis and the power of different classes of molecular markers in discriminating coastal and oceanic Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis> L.) on the Lofoten spawning grounds,Northern Norway

Atlantic cod (Gadus morhua) encompasses many different populations or stocks, which in part are managed separately. In the northeast Atlantic cod is divided into two main management units; northeast Arctic cod and coastal cod. These two groups have traditionally been separated by otolith classification. In this study, the power of different classes of genetic markers in separating the two cod groups was investigated. The variation in thirteen genetic markers, including allozymes, haemoglobin, the scDNA locus Pantophysin (Pan I) and a number of microsatellites was investigated, and mixed stock analysis and individual assignment tests were performed on samples comprising a mixture of individuals of putative coastal and oceanic type cod. The genetic analyses showed a large genetic differentiation between outer stations and stations located closer to the mainland shore. Mixed stock analysis and individual assignment tests used for estimation of stock proportions gave results similar to those obtained by otolith classification. Guest editors: J. Davenport, G. Burnell, T. Cross, M. Emmerson, R. McAllen, R. Ramsay & E. Rogan Challenges to Marine Ecosystems