Microsatellite Allele Frequencies Research Articles

Intraspecific Variation in the Wing Shape and Genetic Differentiation of Reed WarblersAcrocephalus scirpaceusin Croatia

Wing shape plays a major role in determining the aerodynamics and performance of avian flight. Intraspecific variation in wing shape reflects different migratory strategies or distances between breeding and wintering areas. Differences in the wing morphology and genotypes between coastal and inland breeding populations of the Reed Warbler in Croatia were studied. A total of 105 adult birds were measured, while blood samples were taken from 59 individuals. The results showed that the two geographically close populations of Reed Warblers differ in wing morphology, with birds from the inland population having more pointed and more slotted wings than those from the coastal population. There was a slight but significant difference in microsatellite allele frequencies between the two populations (FST = 0.015), suggesting that gene flow between the two populations is partly restricted. The differences in wing morphology imply that the two populations use different migratory routes and/or winter in different areas. Consequently, this migratory pattern may have influenced the contemporary genetic differentiation of the two populations.

Chapter 13: Alaska Song Sparrows (Melospiza Melodia) Demonstrate that Genetic Marker and Method of Analysis Matter in Subspecies Assessments

w e examined genetic and morphological characteristics of the Song Sparrows (Melospiza melodia) of northwestern North America, which have a relatively large number of phe- notypically described subspecies (n = 6 in this region). mitochondrial DNA (mtDNA) sequences showed little information about these subspecies, with no reciprocal monophyly evident. however, differences in body mass and microsatellite allele frequencies supported continued recognition of subspecific units for taxonomy and conservation. Song Sparrow subspecies in this region are probably representative of many recently diverged populations that have not been isolated long enough for complete lineage sorting using mtDNA markers, yet which have evolved differences that are likely to be genetically based. we emphasize the importance of using multiple lines of evidence, genetic and morphological, in assessing subspecific status, lest we overlook important biological diversity that has accrued below the level of full species.

Geographic and genetic boundaries of brown bear (Ursus arctos) population in the Caucasus

The taxonomic status of brown bears in the Caucasus remains unclear. Several morphs or subspecies have been identified from the morphological (craniological) data, but the status of each of these subspecies has never been verified by molecular genetic methods. We analysed mitochondrial DNA sequences (control region) to reveal phylogenetic relationships and infer divergence time between brown bear subpopulations in the Caucasus. We estimated migration and gene flow from both mitochondrial DNA and microsatellite allele frequencies, and identified possible barriers to gene flow among the subpopulations. Our suggestion is that all Caucasian bears belong to the nominal subspecies of Ursus arctos. Our results revealed two genetically and geographically distinct maternal haplogroups: one from the Lesser Caucasus and the other one from the Greater Caucasus. The genetic divergence between these haplogroups dates as far back as the beginning of human colonization of the Caucasus. Our analysis of the least-cost distances between the subpopulations suggests humans as a major barrier to gene flow. The low genetic differentiation inferred from microsatellite allele frequencies indicates that gene flow between the two populations in the Caucasus is maintained through the movements of male brown bears. The Likhi Ridge that connects the Greater and Lesser Caucasus mountains is the most likely corridor for this migration.

Phylogeography of the Japanese pipistrelle bat, Pipistrellus abramus, in China: the impact of ancient and recent events on population genetic structure

The influence of Pleistocene climatic oscillations on shaping the genetic structure of Asian biota is poorly known. The Japanese pipistrelle bat occurs over a wide range in eastern Asia, from Siberia to Japan. To test the relative impact of ancient and more recent events on genetic structure in this species, we combined mitochondrial (cytochrome b) and microsatellite markers to reconstruct its phylogeographic and demographic history on continental China and its offshore islands, Hainan Island and the Zhoushan Archipelago. Our mitochondrial DNA tree recovered two divergent geographical clades, indicating multiple glacial refugia in the region. The first clade was mainly confined to Hainan Island, indicating that gene flow between this population and the continent has been restricted, despite being repeatedly connected to the mainland during repeated glacial episodes. By contrast, haplotypes sampled on the Zhoushan Archipelago were mixed with those from the mainland, suggesting a recent shared history of expansion. Although microsatellite allele frequencies showed clear discontinuities across the sampling range, supporting the current isolation of both Hainan Island and the Zhoushan Archipelago, we also found clear evidence of more recent back colonization, probably via post-glacial expansion or, in the latter case, occasional long distance dispersal. The results obtained highlight the importance of using multiple sets of markers for teasing apart the roles of ancient and more recent events on population genetic structure. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99, 582–594.

Marine landscape shapes hybrid zone in a broadcast spawning bivalve: introgression and genetic structure in Canadian west coast Mytilus

Adult marine mussels are sessile, but their highly dispersive planktonic larval stage plays a critical role in shaping population structure. However, shoreline geography and oceanographic cur- rents can modify the dispersal pattern of pelagic larvae. On Vancouver Island (VI), British Columbia, 3 species of blue mussels (native Mytilus trossulus and introduced M. galloprovincialis and M. edulis) form a localized hybrid zone. Here we genetically mapped the distribution of Mytilus species and pop- ulations along VI and the surrounding islands. Using diagnostic species markers and microsatellite loci, we estimated the extent of the Mytilus hybrid zone on VI and measured population differentiation among the observed sites in 2005 and 2006. We predicted that the distribution of non-native genotypes would be mirrored by the microsatellite allelic patterns, which correspond to oceanographic features that reflect barriers to gene flow in the Strait of Georgia. Generally, non-native genotypes were restricted to southern VI and strong microsatellite population structure was detected. The distribution of non-native genotypes reflected patterns of microsatellite allele frequency in the Strait of Georgia. Using a landscape genetics approach, we identified 2 genetic discontinuities, which correspond to oceanographic and hy- drographic features of the Strait of Georgia. Thus, physical dispersal barriers likely limit the spread of the VI Mytilus hybrid zone; however, additional biological barriers to dispersal must also exist. The VI Mytilus hybrid zone provides an excellent example of complex dispersal patterns in a non-equilibrium system.

Highly Discordant Nuclear and Mitochondrial DNA Diversities in Atka Mackerel

AbstractLevels of neutral genetic diversity largely reflect effective population size (Ne), which is influenced by physical and biological environmental variables. While large populations of marine fishes generally harbor higher diversities than freshwater species, historical demography or the effects of natural selection may significantly reduce Ne and differentially affect genomic diversities. Here, we surveyed levels of genetic diversity and examined genetic structure among populations of the Atka mackerel Pleurogrammus monopterygius across most of its geographic range by screening variation at nine nuclear microsatellite DNA markers (n = 745) and in a 468‐base‐pair segment of the mitochondrial DNA (mtDNA) control region (n = 119). Samples from Japan to the western Gulf of Alaska were collected between 2004 and 2006 at six locations, including temporal replicates at two sites. Microsatellite allele frequency homogeneity across the North Pacific indicated an apparent lack of genetic population substructure. While levels of polymorphism at microsatellite loci were typical for marine fishes (haplotype diversity h = 0.34–0.96), mtDNA control region diversity was extremely low (nucleotide diversity = 0.00007; h = 0.033). Only three mtDNA haplotypes, two occurring as singletons, were detected among 119 individuals. The strong contrast between microsatellite and mtDNA diversities appears to be due to the smaller Ne for mtDNA, perhaps resulting from population bottlenecks during postglacial colonizations of the central North Pacific or the effects of natural selection on mtDNA.

Editorial and retrospective 2010

Editorial and retrospective 2010

The genetic structure of populations of an invading pest fruit fly, Bactrocera tryoni, at the species climatic range limit

Previous population genetic studies of the Queensland fruit fly, Bactrocera tryoni Froggatt (Diptera: Tephritidae), in its central range have shown barely detectable genetic differentiation across distances of almost 3000 km (F(st)=0.003). In this study, we investigated the genetic structuring of southern border populations of B. tryoni, in the region extending from the central population to the recently colonized southern range limit. The expectation was that marginal populations would be small, fragmented population sinks, with local adaptation limited by gene flow or drift. Unexpectedly, we found that the population at the southern extreme of the range was a source population, rather than a sink, for the surrounding region. This was shown by assignment testing of recent outbreaks in an adjoining quarantine area and by indirect migration estimates. Furthermore, populations in the region had formed a latitudinal cline in microsatellite allele frequencies, spanning the region between the central population and the southern range limit. The cline has formed within 250 generations of the initial invasion and appears stable between years. We show that there is restricted gene flow in the region and that effective population sizes are of the order of 10(2)-10(3). Although the cline may result from natural selection, neutral evolutionary processes may also explain our findings.

Patterns of Genetic Variation Among Populations of the Asian Longhorned Beetle (Coleoptera: Cerambycidae) in China and Korea

Abstract Central to the study of invasive species is identifying source populations in their native ranges. Source populations of invasive species can provide important information about species life cycles, host use, and species-specific predators and parasites that could be deployed in a biological control program. The Asian longhorned beetle, Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae), is a serious invasive pest of urban forests in North America and Europe. We analyzed mitochondrial DNA haplotype frequencies and microsatellite allele frequencies to characterize phylogeographical patterns in potential source populations of A.. glabripennis. The data suggest that although there is evidence of genetic structure within and among populations in China, a substantial amount of genetic admixture has occurred. This admixture is probably due to recent reforestation efforts in China where beetles from previously isolated populations moved into new areas of abundant, susceptible, and stressed resources, leading to outbreak populations.

Genetic analyses of the Asian longhorned beetle (Coleoptera, Cerambycidae, Anoplophora glabripennis), in North America, Europe and Asia

The Asian longhorned beetle, (Coleoptera, Cerambycidae, Anoplophora glabripennis (Motschulsky)), is endemic to China and Korea and an important invasive insect in North America and Europe. We analyzed mitochondrial DNA sequence data of invasive populations of A. glabripennis in North America and Europe, and microsatellite allele frequency data of beetles from North America. We show that populations in New York City and Long Island NY; New Jersey, Chicago, IL, and Toronto, Canada have limited genetic diversity compared to populations in China. In addition, the data suggest that separate introduction events were responsible for many of the populations in North America and for European populations in Austria, France, Germany and Italy. Populations on Long Island, NY are suspected to have been initiated by the transport of cut wood from New York City. A. glabripennis beetles found in Jersey City, NJ appear to be derived from an expansion of the New York City, NY population, whereas beetles found in Linden, NJ are an expansion from the Carteret, NJ population. Limited genetic diversity did not stop this invasive insect from establishing damaging populations in North America. Founders of introduced A. glabripennis populations in North America and Europe are likely derived from populations in China that are themselves invasive, rendering difficult the identification of source populations. Invasiveness in an insect’s natural range could be an important predictor of potential pest status of introduced populations.

MICA Gene Polymorphism Not Associated with Nonsyndromic Cleft Lip with or without Cleft Palate in the Japanese Population?

Nonsyndromic cleft lip with or without cleft palate (NSCLP) is one of the most common birth defects. Despite its frequency, the etiology remains largely unknown. Most likely, both genetic and environmental factors contribute to this malformation. A polymorphic gene family, the major histocompatibility complex class I chain-related gene A (MICA), is located about 40 kb centromeric to the HLA-B gene. In this study, we analyzed the association between MICA gene polymorphisms and NSCLP in Japanese patients. The (GCT)n polymorphism of the MICA gene was investigated in 94 patients with NSCLP and 180 normal controls using polymerase chain reaction amplification and denaturing polyacrylamide gel electrophoresis. Our results demonstrate that there are no differences in microsatellite allele frequency between NSCLP patients and controls. However, the microsatellite allele frequency of the MICA-A6 (p = 0.045) allele was increased in male patients, as compared with controls. Further, the MICA-A5 (p = 0.359) allele was also increased in female NSCLP patients. These results suggest that the microsatellite allele frequencies of the MICA-A6 allele increased in male NSCLP patients. Although the MICA-A5 allele increased in female NSCLP patients, the increase was not statistically significant. These results suggest that the MICA gene could be one of the candidate genes for NSCLP.

Analysis of Schistosoma mansoni Population Structure Using Total Fecal Egg Sampling

Many parasite populations are difficult to sample because they are not uniformly distributed between several host species and are often not easily collected from the living host, thereby limiting sample size and possibly distorting the representation of the population. For the parasite Schistosoma mansoni, we investigated the use of eggs, in aggregate, from the stools of infected individuals as a simple and representative sample. Previously, we demonstrated that microsatellite allele frequencies can be accurately estimated from pooled DNA of cloned S. mansoni adults. Here, we show that genotyping of parasite populations from reproductively isolated laboratory strains can be used to identify these specific populations based on characteristic patterns of allele frequencies, as observed by polyacrylamide gel electrophoresis and automated sequencer analysis of fluorescently labeled PCR products. Microsatellites used to genotype aggregates of eggs collected from stools of infected individuals produced results consistent with the geographic distribution of the samples. Preferential amplification of smaller alleles, and stutter PCR products, had negligible effect on measurement of genetic differentiation. Direct analysis of total stool eggs can be an important approach to questions of population genetics for this parasite by increasing the sample size to thousands per infected individual and by reducing bias.

Impacts of supplementation: genetic diversity in supplemented and unsupplemented populations of summer chum salmon (Oncorhynchus keta) in Puget Sound (Washington, USA)

In supplementation programs, hatcheries employ wild-origin fish as brood stock and their offspring are allowed into wild spawning areas. Resource managers use supplementation to support imperiled salmonid populations, seeking to increase census size and possibly effective population size (Ne), while minimizing risks of genetic diversity loss and domestication from hatchery intervention. Here we document impacts of 5–10 years of supplementation on threatened summer-run chum salmon ( Oncorhynchus keta ) in Hood Canal (HC) and Strait of Juan de Fuca (SJF) in Washington State and compare them genetically with unsupplemented summer- and fall-run chum salmon from HC and South Puget Sound. Microsatellite allele frequencies identified four run-timing and geographic groups. HC and SJF summer chum salmon genetic relationships followed a metapopulation pattern of isolation by distance, similar to patterns prior to supplementation, suggesting that supplementation minimally impacted population structure. In most supplemented subpopulations, we detected no effects on diversity and Ne, but high variance in individual pairwise relatedness values indicated over-representation of family groups. In two subpopulations, hatchery impacts (decreased diversity and lower Ne) were confounded with extreme bottlenecks. Rebounds in census sizes in all subpopulations suggest that general survivorship has improved and that possible hatchery effects on genetic diversity will be overcome.

COLOR ASSORTATIVE MATING CONTRIBUTES TO SYMPATRIC DIVERGENCE OF NEOTROPICAL CICHLID FISH

It is still debated vigorously whether sexual selection can result in speciation without physical barriers to gene flow. In this study, we used field data and molecular methods to investigate the gold-normal color polymorphism in two endemic cichlid fish species of crater lake Xiloá, Nicaragua. We found significant assortative mating by color in both Amphilophus xiloaensis and A. sagittae. Focusing on A. xiloaensis, microsatellite allele frequencies, an assignment test, and model-based cluster analysis demonstrates significant and clear genetic differentiation (F(ST)= 0.03) between gold and normal individuals in sympatry. In addition, we find genetic differentiation between all three sympatric and ecologically distinct Midas cichlid species of Lake Xiloá, A. amarillo, A. sagittae, and A. xiloaensis (F(ST)= 0.03 - 0.19), and clear genetic isolation of these species from their closest relative (A. citrinellus) in the neighboring great lake Managua. The A. xiloaensis gold morph is genetically more distinct from the lake's other two Midas cichlid species than is A. xiloaensis-normal. Thus, we have identified sexual isolation based on color that is evident in population genetics and mate choice. Our results suggest that sexual selection through color assortative mating may play an important role in incipient sympatric speciation in Midas cichlids of Nicaragua.

Ecological and genetic relationships of the Forest-M form among chromosomal and molecular forms of the malaria vector Anopheles gambiae sensu stricto.

BackgroundAnopheles gambiae sensu stricto, one of the principal vectors of malaria, has been divided into two subspecific groups, known as the M and S molecular forms. Recent studies suggest that the M form found in Cameroon is genetically distinct from the M form found in Mali and elsewhere in West Africa, suggesting further subdivision within that form.MethodsChromosomal, microsatellite and geographic/ecological evidence are synthesized to identify sources of genetic polymorphism among chromosomal and molecular forms of the malaria vector Anopheles gambiae s.s.ResultsCytogenetically the Forest M form is characterized as carrying the standard chromosome arrangement for six major chromosomal inversions, namely 2La, 2Rj, 2Rb, 2Rc, 2Rd, and 2Ru. Bayesian clustering analysis based on molecular form and chromosome inversion polymorphisms as well as microsatellites describe the Forest M form as a distinct population relative to the West African M form (Mopti-M form) and the S form. The Forest-M form was the most highly diverged of the An. gambiae s.s. groups based on microsatellite markers. The prevalence of the Forest M form was highly correlated with precipitation, suggesting that this form prefers much wetter environments than the Mopti-M form.ConclusionChromosome inversions, microsatellite allele frequencies and habitat preference all indicate that the Forest M form of An. gambiae is genetically distinct from the other recognized forms within the taxon Anopheles gambiae sensu stricto. Since this study covers limited regions of Cameroon, the possibility of gene flow between the Forest-M form and Mopti-M form cannot be rejected. However, association studies of important phenotypes, such as insecticide resistance and refractoriness against malaria parasites, should take into consideration this complex population structure.

Genetic structure of the Eurasian lynx population in north-eastern Poland and the Baltic states

We analyzed the genotypes of Eurasian lynx (Lynx lynx) from three populations in the westernmost part of the species main range. One population was situated at the distribution edge (NE Poland) and the two other (Latvia and Estonia) were located within the main, contiguous range of the species. The aim was to determine if the genetic composition varied among these populations and if there was evidence of isolation among them. Based on microsatellite allele frequencies, we found the allelic richness in Polish lynx to be lower than that in lynx from Latvia and Estonia. We also found significant differentiation among the lynx populations, with the NE Poland population forming a distinct genetic group relative to the two other populations (RST = 0.15 and 0.22, P < 0.0001). We suggest that genetic differentiation among lynx populations is the result of habitat insularisation that limits gene flow. This finding emphasizes the necessity to consider the lynx genetic differentiation in conservation planning of this species in Poland.

High genetic diversity and connectivity in the polyploid invasive seaweedAsparagopsis taxiformis(Bonnemaisoniales) in the Mediterranean, explored with microsatellite alleles and multilocus genotypes

The red alga, Asparagopsis taxiformis, has recently expanded its distribution range into the Western Mediterranean Sea, and populations have now even been found on the Portuguese South coast. All Western Mediterranean populations belong to a single mitochondrial cryptic lineage (referred to as lineage 2 in earlier studies) and probably result from a recent invasion of Indo-Pacific origin. Here we investigate fine-scale population genetic diversity and structure within and among Mediterranean populations of lineage 2 using eight microsatellite loci and compare the obtained patterns with those observed in a Californian population of the same lineage. To generate an appropriate analytical method suitable to the polyploid status of this species, we score microsatellite loci as in a dominant marker system. Thereafter, we produce robust descriptors inferred from frequencies of both microsatellite alleles and multilocus genotypes. Populations from California and the Mediterranean Sea differ considerably in their levels of genetic diversity. In the Mediterranean, populations reproduce predominantly sexually and exhibit high levels of genotypic variation, suggestive either of multiple introductions or of a single introduction by a genetically diverse and large group of individuals. Bayesian clustering revealed one or possibly two weakly supported panmictic subpopulations, indicative of extensive admixture. The expansion of this lineage is rapid, possibly due to the absence of eco-physiological barriers to gene flow throughout its invasive trajectory.

Isolated remnant or recent introduction? Estimating the provenance of Yellingbo Leadbeater's possums by genetic analysis and bottleneck simulation

Effective conservation management requires that genetically divergent populations potentially harbouring important local adaptations be identified and maintained as separate management units. In the case of the endangered Australian Leadbeater's possum (Gymnobelideus leadbeateri), an arboreal marsupial endemic to Victoria, uncertainty over the evolutionary origin of a potentially important extant wild population recently discovered in atypical habitat (lowland swamp) at Yellingbo is hampering such efforts. The population is rumoured to be a recent introduction. Microsatellite allele frequencies at Yellingbo differed substantially from those in sampled populations in montane ash forest (F(ST) between 0.23 and 0.36), and Bayesian clustering analyses of genotypes strongly separated them (K = 2). We conducted a suite of bottlenecking tests which all indicated that Yellingbo had undergone a recent reduction in size. The extent to which the distinctiveness of Yellingbo animals might be expected solely through bottlenecking associated with a recent introduction, was tested by simulating population-history scenarios seeded with genotypes from candidate wild and captive sources. No bottleneck scenario reproduced anything approaching the genetic distinction of the Yellingbo population, with all STRUCTURE analyses placing Yellingbo in a separate cluster to simulated populations (K = 2, minimum F(ST) = 0.13). These results suggest that Yellingbo does not share recent ancestry with other extant populations and instead may be a remnant of an otherwise extinct gene pool. Importantly, this may include genes involved in adaptation to a lowland swamp environment, substantially adding to the conservation importance of this population, and suggesting that separate management may be prudent until evidence suggests otherwise.

Selection and sticklebacks

Over the last decade, there has been increasing circumstantial evidence for the action of natural selection in the genome, arising largely from molecular genetic surveys of large numbers of markers. In nonmodel organisms without densely mapped markers, a frequently used method is to identify loci that have unusually high or low levels of genetic differentiation, or low genetic diversity relative to other populations. The paper by Mäkinen et al. (2008a) in this issue of Molecular Ecology reports the results of a survey of microsatellite allele frequencies at more than 100 loci in seven populations of the three-spined stickleback (Gasterosteus aculeatus). They show that a microsatellite locus and two indel markers located within the intron of the Eda gene, known to control the number of lateral plates in the stickleback (Fig. 1), tend to be much more highly genetically differentiated than other loci, a finding that is consistent with the action of local selection. They identify a further two independent candidates for local selection, and, most intriguingly, they further suggest that up to 15% of their loci may provide evidence of balancing selection.

Extensive polymorphism and geographical variation at a positively selected MHC class II B gene of the lesser kestrel (Falco naumanni)

Understanding the selective forces that shape genetic variation in natural populations remains a high priority in evolutionary biology. Genes at the major histocompatibility complex (MHC) have become excellent models for the investigation of adaptive variation and natural selection because of their crucial role in fighting off pathogens. Here we present one of the first data sets examining patterns of MHC variation in wild populations of a bird of prey, the lesser kestrel, Falco naumanni. We report extensive polymorphism at the second exon of a putatively functional MHC class II gene, Fana-DAB*1. Overall, 103 alleles were isolated from 121 individuals sampled from Spain to Kazakhstan. Bayesian inference of diversifying selection suggests that several amino acid sites may have experienced strong positive selection (omega = 4.02 per codon). The analysis also suggests a prominent role of recombination in generating and maintaining MHC diversity (rho = 4Nc = 0.389 per codon, theta = 0.017 per codon). Both the Fana-DAB*1 locus and a set of eight polymorphic microsatellite markers revealed an isolation-by-distance pattern across the Western Palaearctic (r = 0.67; P = 0.01 and r = 0.50; P = 0.04, respectively). Nonetheless, geographical variation at the MHC contrasts with relatively uniform distributions in the frequencies of microsatellite alleles. In addition, we found lower fixation rates in the MHC than those predicted by genetic drift after controlling for neutral mitochondrial sequences. Our results therefore underscore the role of balancing selection as well as spatial variations in parasite-mediated selection regimes in shaping MHC diversity when gene flow is limited.