[Show abstract][Hide abstract] ABSTRACT: The expression of two or more discrete phenotypes amongst individuals within a species (morphs) provides multiple modes upon which selection can act semi-independently, and thus may be an important stage in speciation. In the present study, we compared two sympatric morph systems aiming to address hypotheses related to their evolutionary origin. Arctic charr in sympatry in Loch Tay, Scotland, exhibit one of two discrete, alternative body size phenotypes at maturity (large or small body size). Arctic charr in Loch Awe segregate into two temporally segregated spawning groups (breeding in either spring or autumn). Mitochondrial DNA restriction fragment length polymorphism analysis showed that the morph pairs in both lakes comprise separate gene pools, although segregation of the Loch Awe morphs is more subtle than that of Loch Tay. We conclude that the Loch Awe morphs diverged in situ (within the lake), whereas Loch Tay morphs most likely arose through multiple invasions by different ancestral groups that segregated before post-glacial invasion (i.e. in allopatry). Both morph pairs showed clear trophic segregation between planktonic and benthic resources (measured by stable isotope analysis) but this was significantly less distinct in Loch Tay than in Loch Awe. By contrast, both inter-morph morphological and life-history differences were more subtle in Loch Awe than in Loch Tay. The strong ecological but relatively weak morphological and life-history divergence of the in situ derived morphs compared to morphs with allopatric origins indicates a strong link between early ecological and subsequent genetic divergence of sympatric origin emerging species pairs. The emergence of parallel specialisms despite distinct genetic origins of these morph pairs suggests that the effect of available foraging opportunities may be at least as important as genetic origin in structuring sympatric divergence in post-glacial fishes with high levels of phenotypic plasticity.
Full-text · Article · Jan 2012 · Biological Journal of the Linnean Society
[Show abstract][Hide abstract] ABSTRACT: Approximately half of the mitochondrial genome inherent within 546 individual Atlantic salmon (Salmo salar) derived from across the species' North Atlantic range, was selectively amplified with a novel combination of standard PCR and pyro-sequencing in a single run using 454 Titanium FLX technology (Roche, 454 Life Sciences). A unique combination of barcoded primers and a partitioned sequencing plate was employed to designate each sequence read to its original sample. The sequence reads were aligned according to the S. salar mitochondrial reference sequence (NC_001960.1), with the objective of identifying single nucleotide polymorphisms (SNPs). They were validated if they met with the following three stringent criteria: (i) sequence reads were produced from both DNA strands; (ii) SNPs were confirmed in a minimum of 90% of replicate sequence reads; and (iii) SNPs occurred in more than one individual.
Pyrosequencing generated a total of 179,826,884 bp of data, and 10,765 of the total 10,920 S. salar sequences (98.6%) were assigned back to their original samples. The approach taken resulted in a total of 216 SNPs and 2 indels, which were validated and mapped onto the S. salar mitochondrial genome, including 107 SNPs and one indel not previously reported. An average of 27.3 sequence reads with a standard deviation of 11.7 supported each SNP per individual.
The study generated a mitochondrial SNP panel from a large sample group across a broad geographical area, reducing the potential for ascertainment bias, which has hampered previous studies. The SNPs identified here validate those identified in previous studies, and also contribute additional potentially informative loci for the future study of phylogeography and evolution in the Atlantic salmon. The overall success experienced with this novel application of HT sequencing of targeted regions suggests that the same approach could be successfully applied for SNP mining in other species.
[Show abstract][Hide abstract] ABSTRACT: Microsatellite genotyping is a common DNA characterization technique in population, ecological and evolutionary genetics research. Since different alleles are sized relative to internal size-standards, different laboratories must calibrate and standardize allelic designations when exchanging data. This interchange of microsatellite data can often prove problematic. Here, 16 microsatellite loci were calibrated and standardized for the Atlantic salmon, Salmo salar, across 12 laboratories. Although inconsistencies were observed, particularly due to differences between migration of DNA fragments and actual allelic size (‘size shifts’), inter-laboratory calibration was successful. Standardization also allowed an assessment of the degree and partitioning of genotyping error. Notably, the global allelic error rate was reduced from 0.05 ± 0.01 prior to calibration to 0.01 ± 0.002 post-calibration. Most errors were found to occur during analysis (i.e. when size-calling alleles; the mean proportion of all errors that were analytical errors across loci was 0.58 after calibration). No evidence was found of an association between the degree of error and allelic size range of a locus, number of alleles, nor repeat type, nor was there evidence that genotyping errors were more prevalent when a laboratory analyzed samples outside of the usual geographic area they encounter. The microsatellite calibration between laboratories presented here will be especially important for genetic assignment of marine-caught Atlantic salmon, enabling analysis of marine mortality, a major factor in the observed declines of this highly valued species.
Electronic supplementary material
The online version of this article (doi:10.1007/s10709-011-9554-4) contains supplementary material, which is available to authorized users.
[Show abstract][Hide abstract] ABSTRACT: Three ecologically and morphologically distinct forms of Arctic charr (Salvelinus alpinus L.) have been identified in Loch Rannoch, Scotland, whose evolutionary status and origins are incompletely understood. A
study was made of restriction fragment length polymorphism (RFLPs) detected variation in the D-loop, ND1 and cytochrome b regions of the mitochondrial genome, encompassing >3500bp. Eight RFLP haplotypes were identified that clustered into three
distinct clans based on restriction differences and into four clans based on sequence differences. Significant differences
in RFLP frequencies were found among all morph groups. The pelagic morph was highly divergent from the two benthic forms,
with the benthic forms having variants from only one genetic clan while the pelagic was dominated by a single variant from
another clan. The relative divergence observed among benthic and pelagic forms is ~10 fold greater when nucleotide divergence
among the haplotypes, as well as haplotype frequency differences, is taken into account. Sequence divergence between haplotypes
in the two main clans is of a similar order to that between haplotypes in these clans and a charr from North America. In contrast,
divergence among the two benthic morphs relates entirely to differences in haplotype frequencies. The study confirms the genetic
distinctiveness of the pelagic and benthic forms as well as of the two benthic forms. It strongly supports previous evidence
that the genetic divergence between the pelagic and benthic populations is allopatric in origin. Additionally, the results
strongly suggest that the two benthic populations have undergone peripatric divergence through the sequential colonisation
of the two basins by one lineage, followed by their spatial separation and reproductive isolation.
[Show abstract][Hide abstract] ABSTRACT: Salmon from geographically representative rivers in North America and Europe were typed for variation at the microsatellite
locus SS1 and the mitochondrial DNA ND-1 restriction site 3971, using PCR amplification and agarose-gel electrophoresis. North American
salmon showed near-fixation for SS1 alleles between 129 and 135 bp in length and the N mtDNA restriction type, while European salmon near-fixation for SS1 alleles between 183 and 219 bp and the A/D mtDNA type. Based on the observed variant frequencies, using these two markers
in combination is predicted to give correct assignment of >99.5% of salmon to continent-of-origin. As both these continental
markers can be screened by agarose-gel electrophoresis, their use offers a more rapid, cheaper, and simpler method for accurate
assignment of Atlantic salmon to continent-of-origin than do existing methods. These markers can be applied to the identification
of salmon in North Atlantic high-seas fisheries, in aquaculture stocks, and in rivers to determine the continent-of-origin
of fish-farm escapes.
Preview · Article · Dec 2005 · ICES Journal of Marine Science
[Show abstract][Hide abstract] ABSTRACT: Small populations may be expected to harbour less genetic variation than large populations, but the relation between census size (N), effective population size (N
e), and genetic diversity is not well understood. We compared microsatellite variation in four small peripheral Atlantic salmon populations from the Iberian peninsula and three larger populations from Scotland to test whether genetic diversity was related to population size. We also examined the historical decline of one Iberian population over a 50-year period using archival scales in order to test whether a marked reduction in abundance was accompanied by a decrease in genetic diversity. Estimates of effective population size (N
e) calculated by three temporal methods were consistently low in Iberian populations, ranging from 12 to 31 individuals per generation considering migration, and from 38 to 175 individuals per generation if they were regarded as closed populations. Corresponding N
e/N ratios varied from 0.02 to 0.04 assuming migration (mean=0.03) and from 0.04 to 0.18 (mean=0.10) assuming closed populations. Population bottlenecks, inferred from the excess of heterozygosity in relation to allelic diversity, were detected in all four Iberian populations, particularly in those year classes derived from a smaller number of returning adults. However, despite their small size and declining status, Iberian populations continue to display relatively high levels of heterozygosity and allelic richness, similar to those found in larger Scottish populations. Furthermore, in the R. Asón no evidence was found for a historical loss of genetic diversity despite a marked decline in abundance during the last five decades. Thus, our results point to two familiar paradigms in salmonid conservation: (1)␣endangered populations can maintain relatively high levels of genetic variation despite their small size, and (2) marked population declines may not necessarily result in a significant loss of genetic diversity. Although there are several explanations for such results, microsatellite data and physical tagging suggest that high levels of dispersal and asymmetric gene flow have probably helped to maintain genetic diversity in these peripheral populations, and thus to avoid the negative consequences of inbreeding.
[Show abstract][Hide abstract] ABSTRACT: PCR primers were successfully designed to amplify small ND1 gene fragments for RFLP genotyping of degraded Atlantic salmon Salmo salar mtDNA. Analysis of archival scales with these primers, when existing primer sets failed, show Atlantic salmon from the George River, Quebec, to include European haplotypes and those from the Kapisidlit River, West Greenland, to be fixed for a European haplotype characteristic of Baltic populations.
No preview · Article · Apr 2005 · Journal of Fish Biology
[Show abstract][Hide abstract] ABSTRACT: Seven novel and highly variable tetranucleotide microsatellite markers, and conditions for multiplexing and simultaneous genotyping six of these in a single run, are described for Atlantic salmon. These provide a highly informative and cost-effective set of molecular markers for genetic studies on cultured and wild populations of the species. The primers sets showed cross-species amplification of appropriately sized amplified products in a number of other salmonid species and suggests the primer sets may have wider application.
No preview · Article · May 2004 · Molecular Ecology Notes
[Show abstract][Hide abstract] ABSTRACT: Matrilineal phylogenetic divergence among Atlantic salmon stocks of the Bay of Fundy in south eastern Canada is investigated. Sequence variation in two regions of the mitochondrial ND1 gene, encompassing 710 base pairs, is described for 168 salmon from 11 rivers. Mean overall haplotype and nucleotide diversity (h and ) observed are 0.5014 and 0.00095, respectively. Nested clade analysis (NCA) and molecular analysis of variance (AMOVA) both point to highly restricted gene flow among rivers and show the haplotype distribution to be geographically structured. Variation among predefined regions of the Bay (16%
) is greater than among populations within these regions (14%
) The main regional differentiation occurs between rivers of the geographically isolated inner Minas Basin and those elsewhere in the Bay. Differentiation most probably reflects the pattern and nature of the historical processes associated with post-glacial colonisation of the area by salmon following the last Pleistocene glacial maximum c. 18000 yrs BP.
[Show abstract][Hide abstract] ABSTRACT: Current understanding of the postglacial colonization of Nearctic and Palearctic species relies heavily on inferences drawn from the phylogeographic analysis of contemporary generic variants. Modern postglacial populations are supposed to be representative of their Pleistocene ancestors, and their current distribution is assumed to reflect the different colonization success and dispersal patterns of refugial lineages. Yet, testing of phylogeographic models against ancestral genomes from glacial refugia has rarely been possible. Here we compare ND1 mitochondrial DNA variation in late Pleistocene (16,000-40,000 years before present), historical and contemporary Atlantic salmon (Salmo salar) populations from northern Spain and other regions of western Europe. Our study demonstrates the presence of Atlantic salmon in the Iberian glacial refugium during the last 40,000 years and points to the Iberian Peninsula as the likely source of the most common haplotype within the Atlantic lineage in Europe. However, our findings also suggest that there may have been significant changes in the genetic structure of the Iberian refugial stock since the last ice age, and question whether modern populations in refugial areas are representative of ice age populations. A common haplotype that persisted in the Iberian Peninsula during the Pleistocene last glacial maximum is now extremely rare or absent from European rivers, highlighting the need for caution when making phylogeographic inferences about the origin and distribution of modern genetic types.
Full-text · Article · Nov 2002 · Molecular Ecology
[Show abstract][Hide abstract] ABSTRACT: The diversity and distribution of mtDNA haplotypes in Atlantic salmon (Salmo salar)L.) from 13 river systems across the species» European range was investigated. Salmon were screened by agarose electrophoresis for variation in a 1400 base pair fragment spanning the ND-1 and 16SrRNA genes. The fragment was amplified by PCR and digested using the restriction endonucleases Ava)II, Dra)I, Hae)III, Hin)fI and Rsa)I. Nine haplotypes were identified and resolved by parsimony analysis into two major clades. Clade I was ubiquitous and predominated in all samples while Clade II was restricted to eight out of 94 individuals in two of the 13 rivers. The first clade shows two sublineages whose frequency distribution is strongly associated with geography. One sublineage dominated in river systems draining into the Baltic sea and in Iceland, and the other in the river systems elsewhere in Europe. No geographical patterns were apparent within these regions but haplotype frequencies among samples, both within and outside the Baltic region, were significantly heterogeneous. Approximately 8% of haplotype frequency variation occurred among samples, 44% between Baltic and non-Baltic samples and 48% within samples. Baltic samples had a significantly lower haplotype and nucleotide diversity than non-Baltic samples. Current and historical factors potentially responsible for the observed levels and distribution of the haplotype variation are discussed.
No preview · Article · Sep 1999 · Biological Journal of the Linnean Society