New polymorphic microsatellite markers of the endangered meadow viper ( Vipera ursinii ) identified by 454 high-throughput sequencing: when innovation meets conservation

The Next Generation Sequencing (pyrosequencing) technique allows rapid, low-cost development of microsatellite markers. We have used this technology to develop 14 polymorphic loci for the endangered meadow viper (Vipera ursinii). Based on 37,000 reads, we developed primers for 66 microsatellite loci and found that 14 were polymorphic. The number of alleles per locus varies from 1 to 12 (for 30 individuals tested). At a cost of about 1/3 that of a normal microsatellite development, we were able to define enough microsatellite markers to conduct population genetic studies on a non-model specie

Low genetic and phenotypic divergence in a contact zone between freshwater and marine sticklebacks: gene flow constrains adaptation

Abstract Background Distinct hybrid zones and phenotypic and genomic divergence is often observed between marine and freshwater threespine sticklebacks (Gasterosteus aculeatus). Nevertheless, cases also exist where marine-freshwater divergence is diffuse despite seemingly similar environmental settings. In order to assess what characterizes these highly different outcomes, we focused on the latter kind of system in the Odder River, Denmark. Here, a previous study based on RAD (Restriction site Associated DNA) sequencing found non-significant genome-wide differentiation between marine and freshwater sticklebacks. In the present study, we analyzed samples on a finer geographical scale. We assessed if the system should be regarded as panmictic, or if fine-scale genetic structure and local selection was present but dominated by strong migration. We also asked if specific population components, that is the two sexes and different lateral plate morphs, contributed disproportionally more to dispersal. Results We assessed variation at 96 SNPs and the Eda gene that affects lateral plate number, conducted molecular sex identification, and analyzed morphological traits. Genetic differentiation estimated by FST was non-significant throughout the system. Nevertheless, spatial autocorrelation analysis suggested fine scale genetic structure with a genetic patch size of 770 m. There was no evidence for sex-biased dispersal, but full-plated individuals showed higher dispersal than low- and partial-plated individuals. The system was dominated by full-plated morphs characteristic of marine sticklebacks, but in the upstream part of the river body shape and frequency of low-plated morphs changed in the direction expected for freshwater sticklebacks. Five markers including Eda were under possible diversifying selection. However, only subtle clinal patterns were observed for traits and markers. Conclusions We suggest that gene flow from marine sticklebacks overwhelms adaptation to freshwater conditions, but the short genetic patch size means that the effect of gene flow on the most upstream region must be indirect and occurs over generations. The occurrence of both weak unimodal and strong bimodal hybrid zones within the same species is striking. We suggest environmental and demographic factors that could determine these outcomes, but also highlight the possibility that long-term population history and the presence or absence of genomic incompatibilities could be a contributing factor

A low-density SNP array for analyzing differential selection in freshwater and marine populations of threespine stickleback (Gasterosteus aculeatus)

BACKGROUND: The threespine stickleback (Gasterosteus aculeatus) has become an important model species for studying both contemporary and parallel evolution. In particular, differential adaptation to freshwater and marine environments has led to high differentiation between freshwater and marine stickleback populations at the phenotypic trait of lateral plate morphology and the underlying candidate gene Ectodysplacin (EDA). Many studies have focused on this trait and candidate gene, although other genes involved in marine-freshwater adaptation may be equally important. In order to develop a resource for rapid and cost efficient analysis of genetic divergence between freshwater and marine sticklebacks, we generated a low-density SNP (Single Nucleotide Polymorphism) array encompassing markers of chromosome regions under putative directional selection, along with neutral markers for background. RESULTS: RAD (Restriction site Associated DNA) sequencing of sixty individuals representing two freshwater and one marine population led to the identification of 33,993 SNP markers. Ninety-six of these were chosen for the low-density SNP array, among which 70 represented SNPs under putatively directional selection in freshwater vs. marine environments, whereas 26 SNPs were assumed to be neutral. Annotation of these regions revealed several genes that are candidates for affecting stickleback phenotypic variation, some of which have been observed in previous studies whereas others are new. CONCLUSIONS: We have developed a cost-efficient low-density SNP array that allows for rapid screening of polymorphisms in threespine stickleback. The array provides a valuable tool for analyzing adaptive divergence between freshwater and marine stickleback populations beyond the well-established candidate gene Ectodysplacin (EDA). ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2164-15-867) contains supplementary material, which is available to authorized users

High Genetic Differentiation Among French Populations of the Orsini's Viper (Vipera ursinii ursinii) Based on Mitochondrial and Microsatellite Data: Implications for Conservation Management

The Orsini's viper (Vipera ursinii) is one of the most threatened snakes in Europe due to its highly fragmented distribution and specific open environment (steppic habitat) requirement. French populations are isolated on top of mountain massifs of the southern Prealps/Alps. Mitochondrial sequences (cytochrome b) and 6 microsatellite loci have been used to estimate the levels of genetic diversity and isolation within and among 11 French fragmented populations (a total of 157 individuals). Eleven cytochrome b haplotypes with a limited divergence were observed (mean divergence between haplotypes: 0.31%). However, we detected considerable genetic differentiation among populations (global FST = 0.76 and 0.26 for mitochondrial and nuclear DNA, respectively). Results indicate that 3 populations possibly went through a bottleneck and 1 population showed low genetic diversity compared with the others. Although a significant isolation by distance was detected for both markers, strong differentiation was also observed between geographically close populations, probably due to the ragged landscape that constitutes a serious barrier to gene flow owing to the limited dispersal capability of the viper. Despite some discrepancies between the 2 markers, 8 Management Units have been identified and should be considered for future management project

A cold-water fish striving in a warming ocean: Insights from whole-genome sequencing of the Greenland halibut in the Northwest Atlantic

Characterizing the extent of genetic differentiation among individuals and its distribution across the genome is increasingly important to inform both conservation and management of exploited species. The Greenland Halibut is one of the main demersal fish species to be commercially exploited in Eastern Canada, and accurate information on geographic population structure and local adaptation is required to ensure the long-term presence of this species. We generated high-quality whole-genome sequencing data for 1,297 Greenland Halibut sampled across 32 locations throughout the Northwest Atlantic (from Arctic Canadian and Greenlandic coasts to the Gulf of St Lawrence). Population genetic structure was analyzed, revealing an absence of population differentiation between Canada and west Greenland but significant genetic differentiation between the Gulf of Saint Lawrence and the remainder of the Northwest Atlantic. Except for Gulf of Saint Lawrence, Greenland Halibut thus appear to be panmictic throughout the Northwest Atlantic. Environmental Association Analyses revealed that the environment explained up to 51 % might be replaced by 51% of the differentiation observed between the two stocks, with both ocean-bottom and surface variables (e.g., temperature and oxygen) involved in the observed genomic differentiation. Altogether, these results indicate that phenotypic differences previously observed between the Gulf of Saint Lawrence and the Northwest Atlantic likely resulted from functional adaptive divergence to their respective environmental conditions. Using coalescent simulations, we also assessed how high levels of migration between the two stocks would allow Greenland Halibut to potentially escape unfavorable environmental conditions in the Gulf of Saint Lawrence. In addition to supporting the management of this important exploited species, this work highlights the utility of using comprehensive genomic datasets to characterize the effects of climate change across a wider range of species

Properties and local environment of p-type dopants and photoluminescent rare-earths implanted into zon single-crystals

Tese de doutoramento em Física, apresentada à Universidade de Lisboa através da Faculdade de Ciências, 2008This thesis presents an experimental study of the local environment of P-type and Rare-Earth dopants implanted in ZnO single-crystals (SCs). Various nuclear and bulk property techniques were combined in the following evaluations: Implantation damage annealing Implantation damage annealing was evaluated in ZnO SCs implanted with Fe, Sr and Ca. P-type dopants Cu and Ag implanted ZnO SCs were studied revealing that the solubility of Cu in substituting Zn is considerably higher than that of Ag. These results are discussed within the scope of the ZnO p-type doping problematic with these elements. Experimental proofs of the As anti-site behavior in ZnO were for the first time attained, i.e., the majority of As atoms are substitutional at the Zn site (SZn), possibly surrounded by two Zn vacancies (VZn). This reinforces the theoretical prediction that As acts as an acceptor in ZnO via the AsZn-2VZn complex formation. The co-doping of ZnO SC with In (donor) and As (acceptor) was addressed. The most striking result is the possible In-As pair formation. Two configurations are proposed for this pair , both with In and As at SZn. A purely In-related defect is also identified. These are preliminary experiments that can serve as guidelines for future and deeper studies. Rare-earths (RE) RE lattice site location, RE surface segregation and optical activation were investigated in Er and Tm implanted ZnO SCs, as a function of annealing temperature, implantation doses and implantation conditions. RE surface segregation and lattice recovery occur for high annealing temperatures, being more pronounced in the low dose implanted crystals. The RE-O clusters formation is suggested for the higher dose implanted samples. Er and Tm optical activation occurs at different annealing temperatures depending on the implantation doses, suggesting that the RE-related emission is associated with defects/impurities in the RE close vicinity that change with the annealings

Data from: The impact of selection, gene flow and demographic history on heterogeneous genomic divergence: threespine sticklebacks in divergent environments

Heterogeneous genomic divergence between populations may reflect selection, but should also be seen in conjunction with gene flow and drift, particularly population bottlenecks. Marine and freshwater threespine stickleback (Gasterosteus aculeatus) populations often exhibit different lateral armor plate morphs. Moreover, strikingly parallel genomic footprints across different marine-freshwater population pairs are interpreted as parallel evolution and gene reuse. Nevertheless, in some geographic regions like the North Sea and Baltic Sea different patterns are observed. Freshwater populations in coastal regions are often dominated by marine morphs, suggesting that gene flow overwhelms selection, and genomic parallelism may also be less pronounced. We used RAD sequencing for analyzing 28,888 SNPs in two marine and seven freshwater populations in Denmark, Europe. Freshwater populations represented a variety of environments: river populations accessible to gene flow from marine sticklebacks and large and small isolated lakes with and without fish predators. Sticklebacks in an accessible river environment showed minimal morphological and genome-wide divergence from marine populations, supporting the hypothesis of gene flow overriding selection. Allele frequency spectra suggested bottlenecks in all freshwater populations, and particularly two small lake populations. However, genomic footprints ascribed to selection could nevertheless be identified. No genomic regions were consistent freshwater-marine outliers, and parallelism was much lower than in other comparable studies. Two genomic regions previously described to be under divergent selection in freshwater and marine populations were outliers between different freshwater populations. We ascribe these patterns to stronger environmental heterogeneity among freshwater populations in our study as compared to most other studies, although the demographic history involving bottlenecks should also be considered in the interpretation of results

phenotypic_data_new

Description of the lateral plate morphology in danish threespine populations. The morph is determined by the number of lateral plates : low plated (10 or fewer plates), partially plated (between 10 and 21 plates) and full plated (more than 21 plates) (Le Rouzic et al. 2011). See the manuscript for more details. Note: For HAL population, the HAL phenotypes are from individuals from the locality HAL reported in the manuscript but not those that were sequenced (due to technical issues