Anthony J. Gharrett

Application of single nucleotide polymorphisms to non‐model species: a technical review

Single nucleotide polymorphisms (SNPs) have gained wide use in humans and model species and are becoming the marker of choice for applications in other species. Technology that was developed for work in model species may provide useful tools for SNP discovery and genotyping in non‐model organisms. However, SNP discovery can be expensive, labour intensive, and introduce ascertainment bias. In addition, the most efficient approaches to SNP discovery will depend on the research questions that the markers are to resolve as well as the focal species. We discuss advantages and disadvantages of several past and recent technologies for SNP discovery and genotyping and summarize a variety of SNP discovery and genotyping studies in ecology and evolution.

Outbreeding depression in hybrids between odd- and even-broodyear pink salmon

Abstract Fewer F 2 hybrids between even- and odd-broodline pink salmon ( Oncorhynchus gorbuscha ), which are lines that are genetically isolated by their strict two-year life cycle, survived than did F 2 controls, indicating outbreeding depression. Cryopreserved sperm of 40 broodyear 1990 males and of 40 broodyear 1991 males fertilized equal subsamples of eggs from 40 broodyear 1992 females. Return rates of F 1 hybrids (1.73%) and controls (1.63%) in 1994 did not differ significantly ( P =0.30). F 2 hybrid and control crosses were made from 40 males and 40 females selected at random from each return group. Offspring were differentially marked and released. In 1996, returns differed significantly ( P =0.011) between hybrids ( n =34, 0.34%) and controls ( n =44, 0.42%). The low rate of return of the control fish was similar to the measured return of a much larger group of tagged Auke Creek pink salmon, and probably not an artifact of the experiment. Although no increase in fluctuating asymmetry of paired meristic counts was observed in either F 1 or F 2 hybrids, size and some meristic counts of hybrids exceed measurements of controls, suggesting heterosis for those traits. The observations of decreased survival in F 2 hybrids confirm previous work [Gharrett, A.J., Smoker, W.W., 1991. Two generations of hybrids between even- and odd-year pink salmon ( O. gorbuscha ). Canadian Journal of Fisheries and Aquatic Science 48(9) 1744–1749]. Although genetic divergence between pink salmon broodlines is large and outbreeding depression might be expected in such unlikely hybrids, the results document the occurrence of outbreeding depression in salmon and signal caution in making management and aquacultural decisions that may create the possibility of outbreeding depression in self-sustaining or cultured populations.

Genetic change for earlier migration timing in a pink salmon population

To predict how climate change will influence populations, it is necessary to understand the mechanisms, particularly microevolution and phenotypic plasticity, that allow populations to persist in novel environmental conditions. Although evidence for climate-induced phenotypic change in populations is widespread, evidence documenting that these phenotypic changes are due to microevolution is exceedingly rare. In this study, we use 32 years of genetic data (17 complete generations) to determine whether there has been a genetic change towards earlier migration timing in a population of pink salmon that shows phenotypic change; average migration time occurs nearly two weeks earlier than it did 40 years ago. Experimental genetic data support the hypothesis that there has been directional selection for earlier migration timing, resulting in a substantial decrease in the late-migrating phenotype (from more than 30% to less than 10% of the total abundance). From 1983 to 2011, there was a significant decrease—over threefold—in the frequency of a genetic marker for late-migration timing, but there were minimal changes in allele frequencies at other neutral loci. These results demonstrate that there has been rapid microevolution for earlier migration timing in this population. Circadian rhythm genes, however, did not show any evidence for selective changes from 1993 to 2009.

Positive Darwinian Selection in the Piston That Powers Proton Pumps in Complex I of the Mitochondria of Pacific Salmon

The mechanism of oxidative phosphorylation is well understood, but evolution of the proteins involved is not. We combined phylogenetic, genomic, and structural biology analyses to examine the evolution of twelve mitochondrial encoded proteins of closely related, yet phenotypically diverse, Pacific salmon. Two separate analyses identified the same seven positively selected sites in ND5. A strong signal was also detected at three sites of ND2. An energetic coupling analysis revealed several structures in the ND5 protein that may have co-evolved with the selected sites. These data implicate Complex I, specifically the piston arm of ND5 where it connects the proton pumps, as important in the evolution of Pacific salmon. Lastly, the lineage to Chinook experienced rapid evolution at the piston arm.

Status of Pacific Salmon and Steelhead Escapements in Southeastern Alaska

Abstract We evaluated the status of Pacific salmon and steelhead (Oncorhynchus mykiss) in southeastern Alaska. Of 9,296 spawning aggregates identified in this region, some data were available for 4,009 (43%), and 928 (10%) had sufficient information to analyze for escapement trend. Of those analyzed, 333 (36%) were increasing, 556 (60%) were stable, 37 (4%) were declining, and 2 (< 1%) showed precipitous declines. We evaluated risk of extinction of spawning aggregates using criteria similar to surveys outside Alaska. We rated 918 (99%) at no or low risk, 8 (∼ 1%) at moderate risk, and 2 (< 1%) at high risk. No spawning aggregates were identified as extinct based on our evaluation of escapement data dating back to 1960. Prior to 1960, two spawning aggregates, one sockeye salmon (0. nerka) and one chum salmon (0. keta), were identified as extinct based on responses to a postal questionnaire. The Alaska Department of Fish and Game grouped spawning aggregates into management units for each species. Management...

Assessment of concordance among genealogical reconstructions from various mtDNA segments in three species of Pacific salmon (genus Oncorhynchus)

Seven segments of mitochondrial DNA (mtDNA), comprising 97% of the mitochondrial genome, were amplified by polymerase chain reaction (PCR) and examined for restriction site variation using 13 restriction endonucleases in three species of Pacific salmon: pink (Oncorhynchus gorbuscha), chum (O. keta) and sockeye (O. nerka) salmon. The distribution of variability across the seven mtDNA segments differed substantially among species. Little similarity in the distribution of variable restriction sites was found even between the mitochondrial genomes of the even‐ and odd‐year broodlines of pink salmon. Significantly different levels of nucleotide diversity were detected among three groups of genes: six NADH‐dehydrogenase genes had the highest; two rRNA genes had the lowest; and a group that included genes for ATPase and cytochrome oxidase subunits, the cytochrome b gene, and the control region had intermediate levels of nucleotide diversity. Genealogies of mtDNA haplotypes were reconstructed for each species, based on the variation in all mtDNA segments. The contributions of variation within different segments to resolution of the genealogical trees were compared within each species. With the exception of sockeye salmon, restriction site data from different genome segments tended to produce rather different trees (and hence rather different genealogies). In the majority of cases, genealogical information in different segments of mitochondrial genome was additive rather than congruent. This finding has a relevance to phylogeographic studies of other organisms and emphasizes the importance of not relying on a limited segment of the mtDNA genome to derive a phylogeographic structure.

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 (∼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.

BACKGROUND MATCHING AND COLOR-CHANGE PLASTICITY IN COLONIZING FRESHWATER SCULPIN POPULATIONS FOLLOWING RAPID DEGLACIATION

Anthropogenic-induced change is forcing organisms to shift their distributions and colonize novel habitats at an increasing rate, which leads to complex interactions among evolutionary processes. Coastrange sculpin (Cottus aleuticus) have colonized recently deglaciated streams of Glacier Bay in Alaska within the last 220 years. We examined divergence among populations in background matching coloration and tested the hypothesis that observed variation is due to morphological color plasticity. To examine how color-change plasticity has interacted with other evolutionary processes, we also determined the influence of colonization on neutral genetic diversity. We observed clinal variation in substrate-matching fish color along the chronological continuum of streams. Microsatellites provided little evidence of genetic subdivision among sculpin populations. Fish color was significantly correlated to substrate color, but was not correlated to neutral population genetic structure. Furthermore, a laboratory experiment revealed that morphological color plasticity could explain much, but not all, of the observed fish color divergence. Our study demonstrates that sculpin in Glacier Bay have colonized and adapted to recently deglaciated habitat and suggests that color change plasticity has aided in this process. This research, therefore, highlights the important role phenotypic plasticity may play in the adaptation of species to rapid climate change.

Characterization of species-specifically amplified SINEs in three salmonid species—Chum salmon, pink salmon, and kokanee: The local environment of the genome may be important for the generation of a dominant source gene at a newly retroposed locus

Short interspersed repetitive elements (SINEs), known as theHpaI family, are present in the genomes of all salmonid species (Kido et al.,Proc. Natl. Acad. Sci. USA 1991, 88: 2326–2330). Recently, we showed that the retropositional efficiency of the SINE family in the lineage of chum salmon is extraordinarily high in comparison with that in other salmonid lineages (Takasaki et al.,Proc. Natl. Acad. Sci. USA 1994, 91: 10153–10157). To investigate the reason for this high efficiency, we searched for members of theHpaI SINE family that have been amplified species-specifically in pink salmon. Since the efficiency of the species-specific amplification in pink salmon is not high and since other members of the same subfamily of SINEs were also amplified species-specifically in pink salmon, the actual sequence of this subfamily might not be the cause of the high retropositional efficiency of SINEs in chum salmon. Rather, it appears that a highly dominant source gene for the subfamily may have been newly created by retroposition, and some aspect of the local environment around the site of retroposition may have been responsible for the creation of this dominant source gene in chum salmon. Furthermore, a total of 11 sequences ofHpaI SINEs that have been amplified species-specifically in three salmon lineages was compiled and characterized. Judging from the distribution of members of the same-sequence subfamily of SINEs in different lineages and from the distribution of the different-sequence subfamilies in the same lineage, we have concluded that multiple dispersed loci are responsible for the amplification of SINEs. We also discuss the additional possibility of horizontal transmission of SINEs between species. The availability of the sets of primers used for the detection of the species-specific amplifications of the SINEs provides a convenient and reliable method for identification of these salmonid species.

Genetic Relationships of Even-Year Northwestern Alaskan Pink Salmon

Abstract Even-year cohorts of northwestern Alaskan pink salmon Oncorhynchus gorbuscha, sampled from 13 Aleutian islands, 1 Kodiak Island, and 4 eastern Bering Sea streams, were surveyed electrophoretically for 29 protein-coding loci. We observed no significant genetic heterogeneity among collections from the Aleutian Islands or within the Bering Sea regions of Bristol Bay and Norton Sound. The Bering Sea and Aleutian Island populations were more closely related to each other than to the Kodiak Island population. This similarity may reflect patterns of post-glacial colonization from the Bering Refuge. Genetic comparisons of northwestern Alaskan pink salmon with Asian populations from Sakhalin Island suggest that the Asian fish are most closely related to the northernmost North American populations that we collected, i.e., those of Norton Sound. Homogeneity among Aleutian Island populations is notable because the islands extend nearly 1,000 km, a distance over which pink salmon populations in other regions ...