Gene–Centromere Distances of Allozyme loci in Even- and Odd-year Pink Salmon, (Oncorhynchus gorbuscha)

We produced gynogenetic progeny families to estimate gene-centromere (G-C) distances of allozyme loci in even-year and odd-year pink salmon (Oncorhynchus gorbuscha). G-C distances of 37 loci distributed on a chromosome ranged from 1 cM at LDH-A1* to 49 cM at ADA-2*, DIA-2*, and sMDH-B1,2*. The distribution of the G-C distances along the chromosome arm was not even and appears telomeric. Eight loci in even-year and seven in odd-year showed high G-C distances (>45 cM), indicating that one crossover per chromosome arm is usual in pink salmon. Variation was observed in the results from different families; 14 loci out of 21 tested, showed heterogeneity. At mAH-3*, G-C distances from five odd-year families ranged from 6 to 37 cM; the widest range observed in this study. At isoloci such as sMDH-A 1,2* and sMDH-B1,2* the distances from different families were grouped into statistically discrete distributions, suggesting that it may be a reflection polymorphism at both isoloci. It appears G-C distances in salmonid species are well conserved with some minor differences.     

Genetic variation of Japanese pink salmon populations inferred from nucleotide sequence analysis of the mitochondrial DNA control region

To estimate genetic variation and structure of pink salmon (Oncorhynchus gorbuscha) populations in Hokkaido, Japan, we analyzed the nucleotide sequence of about 500 bp in a variable portion of the 5′ end of the mitochondrial DNA control region for even- and odd-year broodlines. Sixty-seven haplotypes were detected in the examined individuals. Among these, 25 haplotypes were unique to the even-year broodline, while another 30 haplotypes were unique to the odd-year broodline. Five and three length-heteroplasmic haplotypes were detected in the even-year broodline and odd-year broodline, respectively. The distribution pattern of the 67 haplotypes was different among populations between both broodlines, while not different among populations within the same broodline. The haplotype and nucleotide diversity were higher for even-year broodline populations than for odd-year broodline populations, suggesting greater genetic variation within populations of the even-year broodline. Analysis of molecular variance and pairwise fixation index estimates also demonstrated strong genetic differentiation between even- and odd-year broodlines, although there was no genetic differentiation among populations within the same year broodline. The neutrality tests and mismatch distribution analysis indicate that the demographic history of pink salmon in Japan differs between even- and odd-year populations. Together, these results suggest strong reproductive isolation between the even- and odd-year broodlines of pink salmon, and high gene flow with broodlines due to straying.     

Experimental microevolution: transplantation of pink salmon into the European North

Human-mediated translocations of species beyond their native ranges can enhance evolutionary processes in populations introduced to novel environments. We studied such processes in several generations of pink salmon Oncorhynchus gorbuscha introduced to the European North of Russia using a set of morphological and life-history traits as well as molecular genetic markers with different selective values: protein-coding loci, mtDNA, microsatellites, and MHC. The introduction of reproductively isolated pink salmon broodlines of odd and even years yielded different results. The odd-year broodline established self-reproducing local populations in many rivers of new range, but sustainable changes in external morphology, reproduction, and life-history, as well as the impoverishment of the gene pool occurred. Their successful colonisation of the new range resulted in specialisation manifested in the rapid directional shifts in some highly heritable phenotypic traits accompanied by increased homozygosity at molecular markers as a consequence of genetic drift and selective processes. The returns of transplanted pink salmon of even-year broodline decreased sharply already in the second generation, but there was no marked reduction of genetic diversity. Our data, as well as the analysis of the history of all pink salmon transplantations beyond the species range, demonstrate comparatively greater success of introduced odd-year broodline and permit to assume different adaptive plasticity of the even- and odd-year broodlines in pink salmon, what is most likely determined by differences in their evolutionary histories. Population genetic data suggest that the even-year broodline probably diverged from the odd-year broodline relatively recently and, due to the founder effect, may have lost a part of its genetic variation with which adaptive plasticity potential is associated.     

Genetic stock structure of odd-year pink salmon, Oncorhynchus gorbuscha (Walbaum), from Washington and British Columbia and potential mixed-stock fisheries applications

We used electrophoresis to determine the number and characteristics of genetically distinct stocks of odd-year pink salmon in Washington and southern British Columbia. We analysed 5128 fish from 52 collections (taken in 1985, 1987 and 1989). We observed genetic variation at 53 enzyme-coding loci, 19 of which were polymorphic at the P_o-95 level in at least one stock. Genotypic proportions conformed to Hardy-Weinberg expectations in nearly all cases. The genetic profiles of individual populations were generally stable over the three cycle years studied. Significant differences in allele frequencies at sAAT-3* , PEP-LT* and PGDH* for several stocks were, however, noted between this study and previously reported data for pink salmon. We used G-tests and cluster analysis of genetic distances to evaluate genetic interrelationships among collections and to define genetically distinct stocks. Differentiation among stocks exhibited a clear geographic pattern with three major clusters of stocks recognizable: (1) Hood Canal and Washington Strait of Juan de Fuca stocks, (2) Puget Sound, Fraser River, and southern Canada South Coast stocks, and (3) northern Canada South Coast stocks and Canada North Coast stocks. Computer simulations using 14 and 28 loci, and sample sizes of 15C600, demonstrated that accurate estimates of stock-group composition could be obtained for pink salmon fisheries having a considerable range of stock compositions. The simulations revealed that approximately 50% fewer fish were required to obtain a given level of precision of stock group composition estimates with 28 loci as with the set of 14 loci used in previous investigations.     

Timing of Development During Epiboly in Embryos of Second-Generation Crosses and Backcrosses between Odd- and Even-Broodyear Pink Salmon, Oncorhynchus gorbuscha

Synopsis Odd- and even-year-spawning pink salmon (Oncorhynchus gorbuscha) are genetically isolated; their broodlines differ even in the same natal stream. Hybrids between broodlines exhibit outbreeding depression in survival. Variation in the time to completion of epiboly in embryos appears to be adaptive in both broodlines. We compared stage of development at a time near the completion of epiboly in families of second-generation offspring from crosses between odd- and even-year broodlines with development stages of within-broodyear controls and of backcrossed families. We observed embryos derived from matings of mature fish that were the results of fertilizations made 2 years earlier of eggs from females from the even brood year with semen from males from the even broodyear and with cryopreserved semen from males of the odd broodyear. The resulting fry had been released to the Pacific Ocean and recovered at maturity. Second generation embryos were produced by factorial matings of these mature fish involving (1) female and male controls, (2) female and male hybrids, and (3) both backcrosses. Analysis of variation of development time detected no effect of outbreeding, i.e., differences between controls and second generation hybrids (p > 0.05), but did detect variation between individual female parents (p < 0.03). Neither epistatic nor additive outbreeding depression could be detected in the rate of early embryonic development of pink salmon. However, effects on development rate attributable to female parents indicate that either a maternal effect or early additive genetic effects occurred before the expression of the paternal genome in embryos.     

A linkage map of Atlantic salmon (Salmo salar) reveals an uncommonly large difference in recombination rate between the sexes

A genetic linkage map of the Atlantic salmon (Salmo salar) was constructed, using 54 microsatellites and 473 amplified fragment length polymorphism (AFLP) markers. The mapping population consisted of two full-sib families within one paternal half-sib family from the Norwegian breeding population. A mapping strategy was developed that facilitated the construction of separate male and female maps, while retaining all the information contributed by the dominant AFLP markers. By using this strategy, we were able to map a significant number of the AFLP markers for which all informative offspring had two heterozygous parents; these markers then served as bridges between the male and female maps. The female map spanned 901 cM and had 33 linkage groups, while the male spanned 103 cM and had 31 linkage groups. Twenty-five linkage groups were common between the two maps. The construction of the genetic map revealed a large difference in recombination rate between females and males. The ratio of female recombination rate vs. male recombination rate was 8.26, the highest ratio reported for any vertebrate. This map constitutes the first linkage map of Atlantic salmon, one of the most important aquaculture species worldwide.     

Linkage maps of microsatellite DNA markers for the Pacific oyster Crassostrea gigas

We constructed male and female consensus linkage maps for the Pacific oyster Crassostrea gigas, using a total of 102 microsatellite DNA markers typed in 11-day-old larvae from three families. We identified 11 and 12 linkage groups in the male and female consensus maps, respectively. Alignment of these separate maps, however, suggests 10 linkage groups, which agrees with the haploid chromosome number. The male linkage map comprises 88 loci and spans 616.1 cM, while the female map comprises 86 loci and spans 770.5 cM. The male and the female maps share 74 loci; 2 markers remain unlinked. The estimated coverages for the consensus linkage maps are 79% for the male and 70-75% for the female, on the basis of two estimates of genome length. Ninety-five percent of the genome is expected to lie within 16 and 21 cM of markers on the male and female maps, respectively, while 95% of simulated minimum distances to the male and female maps are within 10.1 and 13.6 cM, respectively. Females have significantly more recombination than males, across 118 pairs of linked markers in common to the parents of the three families. Significant differences in recombination and orders of markers are also evident among same-sex parents of different families as well as sibling parents of opposite sex. These observations suggest that polymorphism for chromosomal rearrangements may exist in natural populations, which could have profound implications for interpreting the evolutionary genetics of the oyster. These are the first linkage maps for a bivalve mollusc that use microsatellite DNA markers, which should enable them to be transferred to other families and to be useful for further genetic analyses such as QTL mapping.     

Estimates of population differentiation in pink salmon Oncorhynchus gorbuscha with using of the microsatellite markers may be underestimated due to high population sizes

Data on the variation at eight microsatellite loci in the Far East salmon Oncorhynchus gorbuscha samples caught in 1984–1985 and 2001–2006 are analyzed. F-statistics indices at all levels of the hierarchical spatial structures are very small. At the same time, the differentiation between populations (according to F _ST estimates) in the odd-year broodline of pink salmon does not exceed the temporal variation within populations. In the even-year broodline, the F-statistics indices at the interregional and intraregional levels are significantly greater than those in the odd-year broodline. F _ST estimates (averaged over the same set of loci) vary widely within the range: the highest values are observed in populations of the coast of North America (except Alaska) and in the new range in the European North of Russia, whereas in the populations of the Asian part of the range and Alaska they are one order of magnitude smaller. The causes of the heterogeneity of the estimates of genetic differentiation within the range and between the broodlines of odd and even years are discussed. Since the mean population size estimates were correlated with the F _ST values, it was assumed that the effect of random genetic drift, the main factor of population divergence in selectively neutral loci, weakens with an increase in the population size. Because of the greater population sizes in pink salmon compared to other salmon species, as well due to the uneven distribution of populations of different size, the usage of microsatellite markers may lead to an underestimation of the true divergence of populations and their regional groups and, consequently, to an overestimation of genetic migration.     

Microsatellite isolation, linkage group identification and determination of recombination frequency in the peach-potato aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae)

Fifteen polymorphic microsatellite markers were used to establish linkage groups and relative rates of recombination in male and female Myzus persicae (Sulzer) (Hemiptera: Aphididae) (peach-potato aphid). We cloned nine markers from M. persicae and for these we report primer sequences and levels of allelic diversity and heterozygosity in four Australian M. persicae populations. Of the remaining six loci, four loci, also cloned from M. persicae, were obtained from G. Malarky (Natural History Museum, London) and two loci from Sitobion miscanthi were used. Additionally, the primer sequences of locus M77, a locus monomorphic in M. persicae but polymorphic in the closely related Myzus antirrhinii, are presented. Eleven of the 15 polymorphic markers were autosomal and four were X-linked. A linkage analysis was performed on a European pedigree of aphids containing five families with between seven and 11 offspring each. There was no linkage between any loci in females. In males, several pairwise comparisons yielded no recombinant offspring. With the exception of locus M40, these observations were supported in a linkage analysis performed on larger families produced from Australian M. persicae crosses. Locus M40 showed segregation consistent with involvement in a translocation between autosomes 1 and 3 in European samples but not in the Australian samples. From the Australian crosses we report an absence of recombination in males but high recombination rates in females. One X chromosome and four autosomal linkage groups were identified and tentatively assigned to chromosomes. The relevance of achiasmate meiosis to the evolution of sex is discussed.     

Outbreeding Depression in Hybrids Between Spatially Separated Pink Salmon, Oncorhynchus gorbuscha, Populations: Marine Survival, Homing ability, and Variability in Family Size

Hybridization between distinct populations and introgression of nonnative genes can erode fitness of native populations through outbreeding depression, either by producing a phenotype intermediate to that of both contributing genomes (and maladapted in either population's environment) or by disrupting distinct coadapted complexes of epistatic genes. In salmon, fitness-related traits such as homing ability or family-size distribution may be eroded. We investigated geographically separated pink salmon populations in repeated trials in independent broodyears (odd and even). Hybrids were made between female Auke Creek (Southeast Alaska) pink salmon and Pillar Creek (Kodiak Island, ～1 000 km away) males; hybrids and their offspring were compared to offspring of control crosses of the same females with Auke Creek males. Parentage assignment from microsatellite analysis was used to improve estimates of survival and straying and to examine variation of family size. Hybridization reduced return rates of adults (a proxy for survival at sea) in the F₁ generation in the odd-year broodline (p < 0.0001) but not in the even-year broodline (p = 0.678). Hybridization reduced survival in both the odd- and even-broodyear F₂ (p < 0.005 and p < 0.0001). Hybridization did not appear to impair homing ability; weekly surveys revealed similar straying rates (～2%) by both hybrid and control fish into nearby (～1 km) Waydelich Creek in both generations in both trials. Hybridization did not increase the index of variability (σ²/μ) in family size. Decreased survival in the hybrid F₂ generation supports an epistatic model of outbreeding depression; nonepistatic effects may have contributed to reduced survival in the odd-broodyear F₁ hybrid fish. Outbreeding depression in hybrids of geographically separated populations demonstrates that introgression of nonnative fish can erode fitness, and should be recognized as a potential detriment of both aquaculture and management practices.     

Genetic divergence in pink salmon introduced into the European north of Russia: microsatellite and allozyme variation analysis

The 1985 introduction into the European North of Russia resulted in the formation of a large stock of pink salmon of the odd-year breeding line. To assess the divergence of the new population and the role of various microevolutionary factors, variation of four microsatellite loci and fifteen genes encoding proteins (allozymes) in samples of fish, running for spawning in rivers of the new area, and in samples from the donor population of the Ola River (Magadan oblast). In the generations 8 and 9 of the introduced pink salmon of the odd-year line, the genetic diversity (the number of alleles and the mean heterozygosity) both at allozyme and at microsatellite loci was significantly lower, than that in the donor population. The explanations of the decline in diversity are discussed. The first evidence for spatial genetic divergence in transplanted fish within the new area has been obtained; the divergence level may be comparable with that characteristic of native populations.     

Genetic divergence in pink salmon introduced into the European North of Russia: Microsatellite and allozyme variation analysis

The 1985 introduction into the European North of Russia resulted in the formation of a large stock of pink salmon of the odd-year broodline. To assess the divergence of the new population and the role of various microevolutionary factors, variation of four microsatellite loci and fifteen genes encoding proteins (allozymes) in samples of fish, running for spawning in rivers of the new area, and in samples from the donor population of the Ola River (Magadan region). In the generations 8 and 9 of the introduced pink salmon of the odd-year line, the genetic diversity (the number of alleles and the mean heterozygosity) both at allozyme and at microsatellite loci was significantly lower, than that in the donor population. The explanations of the decline in diversity are discussed. The first evidence for spatial genetic divergence in transplanted fish within the new area has been obtained; the divergence level may be comparable with that characteristic of native populations.     

Comparative phylogeography of the two pink salmon broodlines: an analysis based on a mitochondrial DNA genealogy

Over most of their natural northern Pacific Ocean range, pink salmon (Oncorhynchus gorbuscha) spawn in a habitat that was repeatedly and profoundly affected by Pleistocene glacial advances. A strictly two-year life cycle of pink salmon has resulted in two reproductively isolated broodlines, which spawn in alternating years and evolved as temporal replicates of the same species. To study the influence of historical events on phylogeographical and population genetic structure of the two broodlines, we first reconstructed a fine-scale mtDNA haplotype genealogy from a sample of 80 individuals and then determined the geographical distribution of the major genealogical assemblages for 718 individuals sampled from nine Alaskan and eastern Asian even- and nine odd-year pink salmon populations. Analysis of restriction site states in seven polymerase chain reaction (PCR)-amplified mtDNA regions (comprising 97% of the mitochondrial genome) using 13 endonucleases resolved 38 haplotypes, which clustered into five genealogical lineages that differed from 0.065 to 0.225% in net sequence divergence. The lineage sorting between broodlines was incomplete, which suggests a recent common ancestry. Within each lineage, haplotypes exhibited star-like genealogies indicating recent population growth. The depth of the haplotype genealogy is shallow ( approximately 0.5% of nucleotide sequence divergence) and probably reflects repeated decreases in population size due to Pleistocene glacial advances. Nested clade analysis (NCA) of geographical distances showed that the geographical distribution observed for mitochondrial DNA (mtDNA) haplotypes resulted from alternating influences of historical range expansions and episodes of restricted dispersal. Analyses of molecular variance showed weak geographical structuring of mtDNA variation, except for the strong subdivision between Asian and Alaskan populations within the even-year broodline. The genetic similarities observed among and within geographical regions probably originated from postglacial recolonizations from common sources rather than extensive gene flow. The phylogeographical and population genetic structures differ substantally between broodlines. This can be explained by stochastic lineage sorting in glacial refugia and perhaps different recolonization routes in even- and odd-year broodlines.     

Gene-centromere mapping of 312 loci in pink salmon by half-tetrad analysis.

We estimated recombination rates between 312 loci and their centromeres in gynogenetic diploid pink salmon (Oncorhynchus gorbuscha) that we produced by initiating development with irradiated sperm and blocking the maternal second meiotic division. Amplified fragment length polymorphisms (AFLPs) were significantly more centromeric than loci identified by three other techniques (allozymes, microsatellites, and PCR using primer sequences from interspersed nuclear elements). The near absence of AFLPs in distal regions could limit their utility in constructing linkage maps. A large proportion of loci had frequency of second division segregation (y) values approaching 1.0, indicating near complete crossover interference on many chromosome arms. As predicted from models of chromosomal evolution in salmonids based upon results with allozyme loci, all duplicated microsatellite loci that shared alleles (isoloci) had y values of nearly 1.0.     

Meiotic recombination in Turnera (Turneraceae): extreme sexual difference in rates, but no evidence for recombination suppression associated with the distyly (S) locus

To explore the rate of recombination resulting from male vs female meiosis, crosses were performed using distylous Turnera subulata as well as a cross involving the introgression of genes from T. krapovickasii into T. subulata. We assayed four loci on the chromosome bearing the S-locus as well as two loci on each of two other linkage groups. Substantial and consistent dimorphism in recombination rates was found with female meiosis resulting in as much as a approximately 6-fold increase relative to male. Aberrant single locus segregation ratios occurred for some loci, particularly when the male (pollen) parent was heterozygous and the cross involved introgressed genes. The extreme trend of greater recombination resulting from female meiosis was, however, maintained in crosses where no aberrant ratios occurred, indicating that the sex dimorphism in recombination is not the result of aberrant segregation. We also exploited this distylous species and tested whether there is recombination suppression around the S-locus because of an inversion or other chromosome rearrangement(s). We found no significant evidence for recombination suppression.     

Twelve loci form a continuous linkage map for human chromosome 18

We have constructed a primary genetic map of human chromosome 18 consisting of 11 DNA markers and one serological marker (JK). Two of these loci define highly polymorphic VNTR systems. The markers define a continuous genetic linkage map of 97 cM in males and 205 cM in females; female genetic distances in a panel of 59 three-generation families were consistently about twice those observed in males. The high odds in support of the linear order of the markers on this recombination map, and the extent of coverage of chromosome 18, indicate that this map will permit efficient linkage studies of human genetic diseases that may be segregating on chromosome 18 and will provide anchor points for development of high-resolution maps for this chromosome.     

Power of QTL mapping experiments in commercial Atlantic salmon populations, exploiting linkage and linkage disequilibrium and effect of limited recombination in males

Whereas detection and positioning of genes that affect quantitative traits (quantitative trait loci (QTL)) using linkage mapping uses only information from recombinants in the genotyped generations, linkage disequilibrium (LD) mapping uses historical recombinants. Thus, whereas linkage mapping requires large family sizes to detect and accurately position QTL, LD mapping is more dependent on the number of families sampled from the population. In commercial Atlantic salmon breeding programmes, only a small number of individuals per family are routinely phenotyped for traits such as disease resistance and meat colour. In this paper, we assess the power and accuracy of combined linkage disequilibrium linkage analysis (LDLA) to detect QTL in the commercial population using simulation. When 15 half-sib sire families (each sire mated to 30 dams, each dam with 10 progeny) were sampled from the population for genotyping, we were able to detect a QTL explaining 10% of the phenotypic variance in 85% of replicates and position this QTL within 3 cM of the true position in 70% of replicates. When recombination was absent in males, a feature of the salmon genome, power to detect QTL increased; however, the accuracy of positioning the QTL was decreased. By increasing the number of sire families sampled from the population to be genotyped to 30, we were able to increase both the proportion of QTL detected and correctly positioned (even with no recombination in males). QTL with much smaller effect could also be detected. The results suggest that even with the existing recording structure in commercial salmon breeding programmes, there is considerable power to detect and accurately position QTL using LDLA.     

Analysis of morphological differencies of the homo- and heterozygous males and females of pink salmon Oncorhynchus gorbuscha (Walbaum) in the population of the river Ola (the northern coast of the Sea of Okhotsk)

The distinctions of the average values and dispersions of the length and the weight of a body in homo-and heterozygotic groupings of five loci (G3PDH*, sMDH-B 1,2*, PGDH*, FDHG* and PGM-2*) were analysed in a population of pink salmon of the river Ola. It was shown that the character of correlation between genotypical structure and morphological trait is not obligatory and identical for each gene. In our opinion polygenic complexes and not separate genes determine the morphological traits.     

A genetic linkage map for coho salmon (Oncorhynchus kisutch)

Construction of genetic linkage maps is an important first step for a variety of genomic applications, such as selective breeding in aquaculture, comparative studies of chromosomal evolution and identification of loci that have played key roles in the evolution of a species. Here we present a sex-specific linkage map for coho salmon. The map was constructed using 148 AFLP markers, 133 microsatellite loci and the phenotypic locus SEX . Twenty-four linkage groups spanning 287.4 cM were mapped in males, and 33 linkage groups spanning 429.7 cM were mapped in females. Several male linkage groups corresponded to two female linkage groups. The combination of linkage groups across both sexes appeared to characterize regions of 26 chromosomes. Two homeologous chromosomes were identified based on information from duplicated loci. Homologies between the coho and rainbow trout maps were examined. Eighty-six loci were found to form common linkage relationships between the two maps; these relationships provided evidence for whole-arm fissions, fusions and conservation of chromosomal regions in the evolution of these two species.     

Application of alternative models to identify QTL for growth traits in an F2 Duroc x Pietrain pig resource population

Background

Several lines of evidence including allozyme analysis, restriction digest patterns and sequencing of mtDNA as well as mini- and micro-satellite allele frequencies indicate that Atlantic salmon (Salmo salar) from North America and Europe are genetically distinct. These observations are supported by karyotype analysis, which revealed that North American Atlantic salmon have 27 pairs of chromosomes whereas European salmon have 29 pairs. We set out to construct a linkage map for a North American Atlantic salmon family and to compare this map with the well developed map for European Atlantic salmon.

Results

We used microsatellite markers, which had previously been mapped in the two Atlantic salmon SALMAP mapping families from the River Tay, Scotland, to carry out linkage analysis in an Atlantic salmon family (NB1) whose parents were derived from the Saint John River stock in New Brunswick, Canada. As large differences in recombination rates between female and male Atlantic salmon have been noted, separate genetic maps were constructed for each sex. The female linkage map comprises 218 markers in 37 linkage groups while the male map has 226 markers in 28 linkage groups. We combined 280 markers from the female and male maps into 27 composite linkage groups, which correspond to the haploid number of chromosomes in Atlantic salmon from the Western Atlantic.

Conclusions

A comparison of the composite NB1 and SALMAP linkage maps revealed the reason for the difference in the chromosome numbers between European and North American Atlantic salmon: Linkage groups AS-4 and AS-32 in the Scottish salmon, which correspond to chromosomes Ssa-6 and Ssa-22, are combined into a single NB1 linkage group as are linkage groups AS-21 and AS-33 (corresponding to chromosomes Ssa-26 and Ssa-28). The comparison of the linkage maps also suggested some additional chromosomal rearrangements, but it will require finer mapping, potentially using SNPs, to test these predictions. Our results provide the first comparison of the genomic architecture of Atlantic salmon from North America and Europe with respect to chromosome organization.