Sequence diversity of Mhc genes in lake whitefish

The sequence variation of three exons of the major histocompatibility complex ( Mhc ) was examined in a lake whitefish Coregonus sp., population from the Swiss lake of Hallwil. DNA sequences from the Mhc class I A1 , A2 and class II B1 exons, corresponding to the α1, α2 and β1 domains of the Mhc glycoproteins, were obtained by the polymerase chain reaction followed by cloning and sequencing. The numbers of variable sequences detected for each exon were 15 ( A1 ), 11 ( A2 ) and 20 ( B1 ). Levels of nucleotide similarity ranged from 82 to 99% for the A1 exon, 58–96% for the A2 and 88–99% for the B1 exon. At the A1 and B1 exons, the nonsynonymous substitution rates ( dn ) exceeded synonymous substitution rates ( ds ) greatly within the peptide binding regions, indicating the effect of balancing selection. Sequence diversity at the A2 exon did not seem to be maintained by balancing selection ( ds > dn ). Phylogenetic comparison of whitefish Mhc sequences with sequences from other salmonid species and more distantly related teleosts indicated shared ancestral (trans-species) polymorphism.     

The major histocompatibility class I locus in Atlantic salmon (Salmo salar L.): polymorphism,linkage analysis and protein modelling

A cDNA library screening using the conserved exon 4 of Atlantic salmon Mhc class I as probe provided the basis for a study on Mhc class I polymorphism in a breeding population. Twelve different alleles were identified in the 82 dams and sires studied. No individual expressed more than two alleles, which corresponded to the diploid segregation patterns of the polymorphic marker residing within the 3'-untranslated tail. Close linkage between the Sasa-UBA and Sasa-TAP2B loci strengthens the claim that Sasa-UBA is the major Mhc class I locus in Atlantic salmon. We found no evidence for a second expressed classical or non-classical Mhc class I locus in Atlantic salmon. A phylogenetic analysis of salmonid Mhc class I sequences showed domains conserved between rainbow trout, brown trout and Atlantic salmon. Evidence for shuffling of the alpha(1) domain was identified and lineages of the remaining alpha(2) through the cytoplasmic tail gene segment can be defined. The coding sequence of one allele was found associated with two different markers, suggesting recombination within the 3'-tail dinucleotide repeat itself. Protein modelling of several Sasa-UBA alleles shows distinct differences in their peptide binding domains and enables a further understanding of the functionality of the high polymorphism.     

Temporal and Spatial Occurrence of Female Chinook Salmon Carrying a Male-Specific Genetic Marker in the Columbia River Watershed

Recent declines in many chinook salmon, Oncorhynchus tshawytscha, populations within the Columbia River watershed have prompted an examination of their reproductive biology. In a previous study many female fall chinook salmon collected in 1999 from the Hanford Reach of the Columbia River tested positive for a male-specific DNA marker (OtY1) found on the Y chromosome. The purpose of this study was to determine if females testing positive for the OtY1 marker could be found in other populations of fall chinook salmon from the Columbia River, and to assess the prevalence of OtY1 incidence in different female cohorts. Post-spawned male and female fall chinook salmon from three different naturally spawning populations (Hanford Reach, Yakima River and Ives Island) and one hatchery population (Priest Rapids Hatchery) on the Columbia River were tested in 2000 and 2001 for the OtY1 marker. Among naturally spawning populations, 57.4% of the females tested positive from the Hanford Reach, 33.3% tested positive from the Yakima River, and 32.5% tested positive from Ives Island. Of the Priest Rapids Hatchery fish, 62.5% of the females tested positive, and significant differences were detected between the 1995–1996 and 1997–1998 female cohorts from this population. No significant differences were detected between any of the female cohorts from the naturally spawning populations. All male chinook salmon samples, tested positive for OtY1.     

Sequence analysis of MHC class I α2 domain exon variants in one diploid and two haploid Atlantic salmon pedigrees

Genetic diversity in the second domain exon of Atlantic salmon ( Salmo salar ) major histocompatibility complex (Mhc) class I was investigated in two dams and nine of their haploid offspring by means of polymerase chain reaction (PCR) and DNA sequence analysis. A similar study was also performed on nine diploid offspring from one of these dams. The complex segregation patterns and sequence similarities between variants make definitive allele, haplotype and locus assignments difficult. There are, however, indications of six Mhc- Sasa class I loci and a fairly well-defined haplotype of four variants. One non-polymorphic variant present in most specimens could be a salmon analogue to the human non-classical loci.     

Screening of Mhc variation in Atlantic salmon (Salmo salar): a comparison of restriction fragment length polymorphism (RFLP), denaturing gradient gel electrophoresis (DGGE) and sequencing

We compared three different molecular methods currently used for screening of Mhc variation in population studies of Atlantic salmon. Restriction fragment length polymorphism (RFLP) of the entire class II gene detected 22 haplotypes. Seventeen exon 2 sequences were obtained from individuals carrying the 22 haplotypes, two of which had not been detected by RFLP. The six alleles (27%) detected by RFLP and not by exon 2 sequencing probably resulted from sequence variation outside exon 2. Within exon 2, RFLP differentiated 88% of the sequences. Alternatively, denaturing gradient gel electrophoresis (DGGE) performed under two run conditions detected 94% of the sequence variation. Both RFLP using different probes, and the two PCR-based methods using three different primer pairs, suggest that there is only a single Mhc class II B gene in the Baltic populations of Atlantic salmon.     

Denaturing gradient gel electrophoresis (DGGE): a rapid and sensitive technique to screen nucleotide sequence variation in populations

We describe a rapid and sensitive method for the detection of nucleotide sequence variation that can be used for large-scale screening of population markers. Denaturing gradient gel electrophoresis (DGGE) detects sequence variants of amplified fragments by the differences in their melting behavior. DGGE detects most single-base substitutions when carried out on products amplified with a primer to which a GC clamp has been added. Although DGGE has been primarily used for the detection of limited numbers of single-base mutations in disease studies, it offers great potential for use in population analysis of genetic markers with greater levels of sequence variation. The methodology described was developed to identify the number and distribution of MHC class I alpha 1 alleles among chinook salmon (Oncorhynchus tshawytscha) populations. DGGE detects 28 of 31 identified alpha 1 sequences, which differ by between 1 and 16 nucleotides and a two-codon indel. By creating a network of control alleles, 22-23 of the MHC alleles can be resolved rapidly and accurately by a single gel run condition, and 27 alleles can be resolved by two gel run conditions. This techniques has been used in surveys scoring alleles from two MHC markers (class I alpha 1 and alpha 2) in 20,000 individuals of chinook and coho (O. kisutch) salmon. A single person in our laboratory now analyzes 160 salmon from one MHC locus per day with DGGE.     

Pinpointing a selective sweep to the chimpanzee MHC class I region by comparative genomics

Chimpanzees experienced a reduction of the allelic repertoire at the major histocompatibility complex (MHC) class I A and B loci, which may have been caused by a retrovirus belonging to the simian immunodeficiency virus (SIV) family. Extended MHC haplotypes were defined in a pedigreed chimpanzee colony. Comparison of genetic variation at microsatellite markers mapping inside and outside the Mhc region was carried out in humans and chimpanzees to investigate the genomic extent of the repertoire reduction. Multilocus demographic analyses underscored that chimpanzees indeed experienced a selective sweep that mainly targeted the chromosomal segment carrying the Mhc class I region. Probably due to genetic linkage, the sweep also affected other polymorphic loci, mapping in the close vicinity of the Mhc class I region genes. Nevertheless, although the allelic repertoire at particular Mhc class I and II loci appears to be limited, naturally occurring recombination events allowed the establishment of haplotype diversity after the sweep. However, recombination did not have sufficient time to erase the signal of the selective sweep.     

Effects of natural selection on patterns of DNA sequence variation at the transferrin, somatolactin, and p53 genes within and among chinook salmon (Oncorhynchus tshawytscha) populations

This paper describes DNA sequence variation within and among four populations of chinook salmon (Oncorhynchus tshawytscha) at the transferrin, somatolactin and p53 genes. Patterns of variation among salmon species at the transferrin gene have been hypothesized to be shaped by positive natural selection for new alleles because the rate of nonsynonymous substitution is significantly greater than the rate of synonymous substitution. The twin goals of this study were to determine if the history of selection among salmon species at the transferrin gene is also reflected in patterns of intraspecific variation in chinook salmon, and to look for evidence of local adaptation at the transferrin gene by comparing patterns of nonsynonymous and synonymous variation among chinook salmon populations. The analyses presented here show that unlike patterns of variation between species, there is no evidence of greater differentiation among chinook salmon populations at nonsynonymous compared to synonymous sites. There is also no evidence of a reduction of within-species variation due to the hitchhiking effect at the transferrin gene, although in some populations nonsynonymous and synonymous derived mutations are both at higher frequencies than expected under a simple neutral model. Population size weighted selection coefficients (4Ns) that are consistent with both the inter and intraspecific data range from approximately 10 to approximately 235, and imply that between 1 and 40% of new nonsynonymous mutations at the transferrin gene have been beneficial.     

Regional differences in responses of chinook salmon populations to large-scale climatic patterns

Aim To quantitatively explore the extent to which many different populations of the same species (chinook salmon, Oncorhynchus tshawytscha) respond cohesively to a common large‐scale climatic trend. Location The Columbia River basin of the northwestern US. Methods I used regression analyses to describe the downward trend in population growth (number of recruits per spawning adult) for thirteen populations of chinook salmon distributed among three geographical regions: Snake River, Upper Columbia River and Middle Columbia River. I then used residuals from these regressions to characterize per capita productivity for each brood year. Positive residuals indicated productivity higher than that predicted by the time series, while negative residuals revealed years in which productivity was lower than predicted. I next used analysis of covariance (ancova ) to test the null hypothesis that associations between ocean/climate conditions and deviations from predicted population growth did not vary among geographical regions. All ancova s used residuals generated from the regressions as the response variable, geographical region as the main effect, and climatic condition [characterized by the Pacific Decadal Oscillation index (PDO)] as the covariate. A major climate shift occurred in 1977, and because the association of the PDO with salmon productivity varied between the pre‐ and post‐1977 climate regimes, I analysed data from the two regimes separately. Results There were marked impacts of climate on salmon production that varied among geographical regions and between decade‐scale climate regimes. During the pre‐1977 climate regime, productivity of salmon populations from the Snake River tended to exceed expectations (i.e. residuals were positive) when values of the PDO were negative. In contrast, this pattern was not evident in populations from the upper or middle Columbia Rivers. During the post‐1977 regime when ocean productivity was generally lower, the association of the PDO with salmon productivity changed – productivity tended to fall short of expectations (i.e. residuals were negative) when values of the PDO were negative. Main conclusions Understanding the linkages between salmon populations and climate is critical as managers attempt to preserve threatened salmon populations in the face of both natural or human‐induced climate variation and the litany of human activities affecting salmon. An important step in this understanding is the recognition that the response to ocean/climate change by salmon populations of the same species and river basin is not necessarily homogeneous.     

Environmental correlates of life-history variation in juvenile chinook salmon, Oncorhynchus tshawytscha (Walbaum)

Throughout its native North Pacific, the chinook salmon, Oncorhynchus tshawytscha (Walbaum), exists as twolife-history types that aredistinguished by the age at which juvenile salmon migrate to sea as smolts. 'Stream-type' chinook migrate seaward after I or more years of feeding in fresh water, whereas 'ocean-type' fish migrate to sea as newly emerged fry or after 2–3 months in fresh water. Stream-type chinook predominate in populations distant from the sea south of 56° N, and in both inland and coastal populations north of this point. By contrast, ocean-type chinook predominate in coastal populations south of 56° N, but are rare in populations in more northerly latitudes. Stream-type populations are associated with areas of low 'growth opportunity' (as measured by temperature and photoperiod regimes) and/or areas distant from the sea compared to ocean-type. Geographic variability in juvenile life history is suggested to result, in part, from environmental modulation of smolting timing via differences in growth opportunity among geo-graphic areas. In addition, differences in migration distance and temperature regime may result in selection for different sizes at migration among populations which, through differences in growth opportunity, might promote geographic variability in age at seaward migration.     

Genetic variation in survival of chinook salmon (Oncorhynchus tshawytscha) alevins exposed to an unidentified agent of mortality

Mortality of an unknown etiology occurs after hatching and before emergence among Harrison River chinook salmon (Oncorhynchus tshawytscha) alevins incubated in the Chehalis River Hatchery, British Columbia. Inter- and intra-stock genetic variation for alevin survival and time to death was investigated at Chehalis Hatchery in factorial crosses among chinook salmon from the Harrison and Capilano rivers. Alevin survival by family ranged from 0 to 100%, with a mean value of 35.2%. The mean family survival of pure Harrison alevins (13.0%) was significantly lower than that of Capilano alevins (64.1%). For the Harrison stock, estimates of the heritability of survival were 1.05 +/- 0.62 (sire component) and 0.03 +/- 0.07 (dam component). For the Capilano stock, the corresponding estimates were 0.79 +/- 0.53 and 0.80 +/- 0.54. Family means of time to death ranged from 7.5 to 48 days after exposure to mortality-inducing agents. The mean times to death for pure Harrison (15.3 days) and Capilano (21.8 days) families were not significantly different. Sire and dam component heritability estimates for time to death were high for the Harrison stock (1.39 +/- 0.87 and 0.71 +/- 0.46) but low for the Capilano stock (0.06 +/- 0.11 and 0.17 +/- 0.18). Values of survival and time to death for the reciprocal interstock hybrid alevins generally fell between those of the parental stocks. Neither survival nor time to death differed significantly between the reciprocal hybrids, but both traits were more strongly influenced by sire than by dam. The possibility of asynchronous paternal and maternal allele activation during embryonic development was proposed as an explanation for the strong paternal effects observed in this study.     

Sequence analysis of MHC class I α2 from sockeye salmon (Oncorhynchus nerka)

Most studies assessing adaptive MHC diversity in salmon populations have focused on the classical class II DAB or DAA loci, as these have been most amenable to single PCR amplifications due to their relatively low level of sequence divergence. Herein, we report the characterization of the classical class I UBA α2 locus based on collections taken throughout the species range of sockeye salmon (Oncorhynchus nerka). Through use of multiple lineage-specific primer sets, denaturing gradient gel electrophoresis and sequencing, we identified thirty-four alleles from three highly divergent lineages. Sequence identity between lineages ranged from 30.0% to 56.8% but was relatively high within lineages. Allelic identity within the antigen recognition site (ARS) was greater than for the longer sequence. Global positive selection on UBA was seen at the sequence level (dN:dS = 1.012) with four codons under positive selection and 12 codons under negative selection.     

Genetic Variation within and Between Domesticated Chinook Salmon, Oncorhynchus tshawytscha, Strains and their Progenitor Populations

Domesticated chinook salmon strains in British Columbia (BC), Canada are believed to have originated primarily from populations of the Big Qualicum (BQ) River and Robertson Creek (RC) on Vancouver Island in the early 1980s. The number of parental fish that gave rise to the domesticated strains and their subsequent breeding history during approximately five ensuing generations of domestication were not documented. Genetic variation at 13 microsatellite loci was examined in samples from two domesticated strains and the two progenitor populations to determine the genetic relationships among them. The domesticated strains had lower allelic diversity and tended to have lower levels of expected heterozygosity than did the BQ and RC progenitor populations. Only three alleles over all 13 loci were detected in the domesticated strains that were not present in the BQ and RC samples, whereas the progenitor strains possessed over 25 (BQ) and 43 (RC) private alleles. Genetic distance and F_ST values also indicated a closer relationship of the domesticated strains with the BQ than the RC population. One domesticated strain had a significant excess of heterozygosity compared with that expected under conditions of mutation-drift equilibrium, indicative of a recent genetic bottleneck. Genetic differentiation between the domesticated strains was as great as that distinguishing them from the progenitor populations, indicating that the genetic base of domesticated chinook salmon could be increased by hybridization. The existence of genetically distinct domesticated strains of chinook salmon in coastal BC generates the need for an evaluation of potential genetic interactions between domesticated escapees and natural spawning populations.     

Major histocompatibility complex differentiation in Sacramento River chinook salmon

The chinook salmon of the Sacramento River, California, have been reduced to a fraction of their former abundance because of human impact and use of the river system. Here we examine the genetic variation at a major histocompatibility complex class II exon in the four Sacramento chinook salmon runs. Examination of the alleles found in these and other chinook salmon revealed nucleotide patterns consistent with selection for amino acid replacement at the putative antigen-binding sites. We found a significant amount of variation in each of the runs, including the federally endangered winter run. All of the samples were in Hardy-Weinberg proportions. A significant amount of genetic differentiation between runs was revealed by several measures of differentiation. Winter run was the most genetically divergent, while the spring, late-fall, and fall runs were less differentiated.     

Distribution,prevalence and severity of Parvicapsula minibicornis infections among anadromous salmonids in the Fraser River,British Columbia,Canada

Polymerase chain reaction (PCR) and microscopic examination of stained kidney sections were used to diagnose infections with the myxozoan parasite Parvicapsula minibicornis in maturing Fraser River salmon. In 2 series of collections, the parasite was detected in 109 of 406 migrating sockeye salmon Oncorhynchus nerka belonging to Early Stuart, Early Summer and Summer run-timing groups, mainly upper Fraser River stocks. However, the parasite was detected neither in fish at sea nor once they had migrated several 100 km upstream. Prevalence then increased to 95% or greater at the spawning grounds. Histological examination of kidney was less sensitive than PCR in detecting the parasite in salmon collected from the earliest sites in both collections found positive by PCR. Severity of infection was greatest at the spawning grounds. Development of infection in sockeye, measured by prevalence, severity or by the rate of false-negative histological diagnoses, appeared to be a useful estimate of in-river residence time. Prevalence and severity of infections in sequential samples of Harrison River and Weaver Creek sockeye stocks collected from the Harrison River indicated that more time had elapsed since parasite transmission than would be predicted based on migration distance alone. Pink salmon Oncorhynchus gorbuscha, coho salmon O. kisutch and chinook salmon O. tshawytscha were found to be infected with the parasite. Development of P. minibicornis in pink salmon was most similar to that in sockeye. Pink and coho salmon may be at risk to the pathological consequences of P. minibicornis infection.     

Persistence of Mhc heterozygosity in homozygous clonal killifish,Rivulus marmoratus: implications for the origin of hermaphroditism

The mangrove killifish Rivulus marmoratus, a neotropical fish in the order Cyprinodontiformes, is the only known obligatorily selfing, synchronous hermaphroditic vertebrate. To shed light on its population structure and the origin of hermaphroditism, major histocompatibility complex (Mhc) class I genes of the killifish from seven different localities in Florida, Belize, and the Bahamas were cloned and sequenced. Thirteen loci and their alleles were identified and classified into eight groups. The loci apparently arose approximately 20 million years ago (MYA) by gene duplications from a single common progenitor in the ancestors of R. marmoratus and its closest relatives. Distinct loci were found to be restricted to different populations and different individuals in the same population. Up to 44% of the fish were heterozygotes at Mhc loci, as compared to near homozygosity at non-Mhc loci. Large genetic distances between some of the Mhc alleles revealed the presence of ancestral allelic lineages. Computer simulation designed to explain these findings indicated that selfing is incomplete in R. marmoratus populations, that Mhc allelic lineages must have diverged before the onset of selfing, and that the hermaphroditism arose in a population containing multiple ancestral Mhc lineages. A model is proposed in which hermaphroditism arose stage-wise by mutations, each of which spread through the entire population and was fixed independently in the emerging clones.     

The road to extinction is paved with good intentions: negative association of fish hatcheries with threatened salmon

Hatchery programmes for supplementing depleted populations of fish are undergoing a worldwide expansion and have provoked concern about their ramifications for populations of wild fish. In particular, Pacific salmon are artificially propagated in enormous numbers in order to compensate for numerous human insults to their populations, yet the ecological impacts of this massive hatchery effort are poorly understood. Here we test the hypothesis that massive numbers of hatchery-raised chinook salmon reduce the marine survival of wild Snake River spring chinook, a threatened species in the USA. Based on a unique 25-year time-series, we demonstrated a strong, negative relationship between the survival of chinook salmon and the number of hatchery fish released, particularly during years of poor ocean conditions. Our results suggest that hatchery programmes that produce increasingly higher numbers of fish may hinder the recovery of depleted wild populations.     

Allelic and haplotypic diversity at the major histocompatibility class II within domesticated Australian Atlantic salmon (Salmo salar L.)

Variation within the major histocompatibility (MH) class II alpha gene ( Sasa-DAA ) was compared between domesticated Australian Atlantic salmon and their ancestral Canadian population. The level of Sasa-DAA and MH class II beta gene ( Sasa-DAB ) sequence variation was also examined within the Australian population and compared with that published for European Atlantic salmon populations. In contrast to variation previously reported for non-coding microsatellite loci, a high level of MH class II allelic variation has been maintained within the domesticated Australian populations. Furthermore, a high level of Sasa-DAA and Sasa-DAB allele sequence diversity was also observed and exceeded that reported for other cultured Atlantic salmon populations. The number of Sasa-DAB allele sequences (14) surpassed the number of Sasa-DAA allele sequences (9) to produce 14 unique class II haplotypes. We conclude that the Australian Atlantic salmon populations show high MH class II allelic and haplotypic variation compared with both its ancestral Canadian population and other cultured Atlantic salmon populations.     

Comparative genomic analysis of two avian (quail and chicken) MHC regions

We mapped two different quail Mhc haplotypes and sequenced one of them (haplotype A) for comparative genomic analysis with a previously sequenced haplotype of the chicken Mhc. The quail haplotype A spans 180 kb of genomic sequence, encoding a total of 41 genes compared with only 19 genes within the 92-kb chicken Mhc. Except for two gene families (B30 and tRNA), both species have the same basic set of gene family members that were previously described in the chicken "minimal essential" Mhc. The two Mhc regions have a similar overall organization but differ markedly in that the quail has an expanded number of duplicated genes with 7 class I, 10 class IIB, 4 NK, 6 lectin, and 8 B-G genes. Comparisons between the quail and chicken Mhc class I and class II gene sequences by phylogenetic analysis showed that they were more closely related within species than between species, suggesting that the quail Mhc genes were duplicated after the separation of these two species from their common ancestor. The proteins encoded by the NK and class I genes are known to interact as ligands and receptors, but unlike in the quail and the chicken, the genes encoding these proteins in mammals are found on different chromosomes. The finding of NK-like genes in the quail Mhc strongly suggests an evolutionary connection between the NK C-type lectin-like superfamily and the Mhc, providing support for future studies on the NK, lectin, class I, and class II interaction in birds.