José-Luis García-Marín

Postglacial colonization of brown trout in Europe based on distribution of allozyme variants

Compatible data from 11 polymorphic allozyme loci were assembled from 14 published studies and additional unpublished information for 232 native collections of brown trout sampled over a broad geographical range. Most of the genetic variation was explained by the distribution of allelic variation at the LDH-C* and CK-A1* loci. Patterns of geographical distribution coupled with postglacial geological events supported a model of colonization from three preglacial lineages, each fixed for different sets of alleles at these two loci. During glacial retreat, recolonization is proposed to have occurred mainly into adjacent areas through (i) a north-western migration from an eastern Mediterranean–Caspian refuge, (ii) a northern expansion from a refuge in Atlantic drainages of Iberia and southern France, and (iii) a northern and eastern radiation from a refuge centred near the English Channel. Extant populations in deglaciated areas are suggested to represent mixed or pure descendants of these migrating groups. Repopulation from a fourth Mediterranean refuge distinguished by the presence of the LDH-A2*100QL allele was excluded based on the absence of this allele in repopulated areas.

Efficiency of markers and methods for detecting hybrids and introgression in stocked populations

AbstractDetection of hybridization and introgression in wild populations that have been supplemented by hatchery fish is necessary during development of conservation and management strategies. Initially, allozyme data and more recently highly polymorphic microsatellite markers have been used to obtain this information. We used both markers to assess the effectiveness of four assignment methods (Structure, NewHybrids, Baps and GeneClass) to detect hatchery introgression in wild stocked populations. Simulations of hybrid genotypes from real parental data revealed that the number and type of markers used with Structure, NewHybrids and Baps can identify as admixed most first and second generation hybrids as well as first generation backcrosses. In wild populations, introgression rates estimated from different markers and methods were correlated. However, slight disagreements were observed at both population and individual levels. Overall, the fully Bayesian (Structure, NewHybrids and Baps) performed better than partially Bayesian (GeneClass) assignment tests. In wild collections, Baps analyses were limited because of the lack of a native baseline. In all cases, the efficiency of methods was reduced as introgression increased.

Origin and genetic diversity of mosquitofish ( Gambusia holbrooki ) introduced to Europe

We provide mitochondrial sequence variation of the invasive fish Gambusiaholbrooki from 24 European populations, from Portugal to Greece. Phylogeographic structure in Europe was compared with genetic data from native samples (USA) and historical records were reviewed to identify introduction routes. Overall, data agree with records of historical introductions and translocations, and indicate that the most abundant haplotype throughout Europe originated from North Carolina and corresponded to the first introduction in 1921 to Spain, being transferred to Italy in 1922 and to many countries afterwards. Our results also show that at least another independent introduction occurred first in France and subsequently from France to Greece. Haplotypes of G. affinis were not detected in our European sampling effort but historical records and other data suggest that this species was introduced to Italy in 1927 and it might be present. At the continental scale, there is less diversity in Europe than in North America, in agreement with the low number of introduced fish. At the local scale, some European populations gained diversity from multiple introductions and from “de novo” mutations.

Gene flow and maintenance of genetic diversity in invasive mosquitofish (Gambusia holbrooki).

Genetic analyses contribute to studies of biological invasions by mapping the origin and dispersal patterns of invasive species occupying new territories. Using microsatellite loci, we assessed the genetic diversity and spatial population structure of mosquitofish (Gambusia holbrooki) that had invaded Spanish watersheds, along with the American locations close to the suspected potential source populations. Mosquitofish populations from the Spanish streams that were studied had similar levels of genetic diversity to the American samples; therefore, these populations did not appear to have undergone substantial losses of genetic diversity during the invasion process. Population structure analyses indicated that the Spanish populations fell into four main clusters, which were primarily associated with hydrography. Dispersal patterns indicated that local populations were highly connected upstream and downstream through active dispersal, with an average of 21.5% fish from other locations in each population. After initially introducing fish to one location in a given basin, such dispersal potential might contribute to the spread and colonization of suitable habitats throughout the entire river basin. The two-dimension isolation-by-distance pattern here obtained, indicated that the human-mediated translocation of mosquitofish among the three study basins is a regular occurrence. Overall, both phenomena, high natural dispersal and human translocation, favor gene flow among river basins and the retention of high genetic diversity, which might help retain the invasive potential of mosquitofish populations.

Genetic characterization of the invasive mosquitofish (Gambusia spp.) introduced to Europe: Population structure and colonization routes

Biological invasions are considered one of the main anthropogenic factors that reduce the abundance of native species. Understanding the patterns of population structure and behavior of introduced species is important to determine invasion sources and pathways, in addition to improving the protective management of native species. Thus, we set out to advance our knowledge about the mosquitofish Gambusia spp., which is an invasive species that was introduced to southern Europe in 1921 to control mosquito populations. We assessed the genetic diversity and population structure of this species at 13 European locations, by screening variation at six microsatellite loci. We also evaluated six American samples (four of G. holbrooki and two of G. affinis) to identify the most likely source of the populations that established in Europe, and to determine whether G. affinis is also present. The results showed that, while there was evidence of recent bottleneck events in a few isolated locations, most introduced populations harbored a considerable amount of gene diversity, probably because of multiple introductions and secondary contacts. Populations displayed strong genetic differentiation that was mainly associated with geographical distance. At least two main routes of colonization of G. holbrooki seem to have occurred in Europe. The first, and more ancient colonization, was consistent with historical records, with the species invading the Iberian Peninsula. A second and more recent colonization probably occurred in Greece and, from there, France. The presence of G. affinis was not detected in any of the European samples.

Population and family structure of brown trout, Salmo trutta, in a Mediterranean stream

The physical arrangement of closely related individuals is expected to significantly influence the pattern of population genetic structure. For example, if related individuals are non-randomly distributed and included in samples, this may lead to exaggerated conclusions about genetic differentiation. In the present study, we compared population structure v. family relationships of brown trout (Salmo trutta L.) along a Mediterranean stream (Pyrenees) by using eight microsatellite loci. Results showed low levels of genetic (FST) differentiation between collections in a 6.5-km transect along the stream, and a significant correlation between genetic and geographical distance matrices, indicating a weak population structure associated with spatial distribution. Our data also indicated that geographical proximity of related individuals in the youngest (0þ ,1 þ) cohorts probably was associated with limited dispersal of younger brown trout from spawning redds. Family relationships provided evidence, however, for movement of adult trout over distances of a few kilometres that probably contributed to the low observed differentiation. Dispersal of adult Mediterranean trout contrasts with the clustering of related older trout observed for some north European rivers and could be related to the reduced productivity in southern stream basins.

Genomic survey provides insights into the evolutionary changes that occurred during European expansion of the invasive mosquitofish (Gambusia holbrooki)

Biological invasions rank among the main global threats for biodiversity. The Eastern mosquitofish (Gambusia holbrooki) is considered one of the 100 world worst invasive species due to its high adaptation capability to new environments. Using the restriction‐site‐associated DNA tags (RADtags), introduced European locations were compared against native US mosquitofish populations to analyse genomic changes that occurred during invasive process of European locations. After filtering, 7724 RADtags containing only one SNP were retained for population studies. Comparative genomics indicated that 186 of these RADtags matched sequences in the transcriptome of Xyphophorus maculatus, the most closely related genome available. Genomic analyses showed that invasive populations show high reductions in diversity. Further, analyses of population structuring based on these data are concordant with previous analyses based on microsatellites. It is concluded that during the invasion process genetic drift was the main evolutionary force affecting patterns of diversity and population structure. While recognizing that positive selection could be masked by the strong drift during founder events, adaptive processes were evidenced in a reduced number of RADtags (<2%), with only one of these in a putative coding region. Surprisingly, balancing selection was detected in several coding RADtags, suggesting that the preservation of polymorphism in specific genes could be more important than the average population diversity for the population maintenance at any location, particularly for the survival of introduced populations.

Identification and conservation of remnant genetic resources of brown trout in relict populations from Western Mediterranean streams

Brown trout is a cold-adapted freshwater species with restricted distribution to headwater streams in rivers of the South European peninsulas, where populations are highly vulnerable because Mediterranean regions are highly sensitive to the global climatic warming. Moreover, these populations are endangered due to the introgressive hybridization with cultured stocks. Individuals from six remnant populations in Western Mediterranean rivers were sequenced for the complete mitochondrial DNA control region and genotyped for 11 nuclear markers. Three different brown trout lineages were present in the studied region. Significant genetic divergence was observed among locations and a strong effect of genetic drift was suggested. An important stocking impact (close to 25%) was detected in the zone. Significant correlations between mitochondrial-based rates of hatchery introgression and water flow variation suggested a higher impact of stocked females in unstable habitats. In spite of hatchery introgression, all populations remained highly differentiated, suggesting that native genetic resources are still abundant. However, climatic predictions indicated that suitable habitats for the species in these rivers will be reduced and hence trout populations are highly endangered and vulnerable. Thus, management policies should take into account these predictions to design upstream refuge areas to protect remnant native trout in the region.

Phylogenetic diversity within the endemic brown trout Duero lineage: implications for conservation and management

Brown trout display great phenotypic and genetic variability. Use of mitochondrial DNA (mtDNA) variation has allowed the definition of seven different lineages in this species to date. One of them, the Duero (DU) lineage, was initially detected in the inner section of the Duero River in Spain, where it showed a parapatric distribution with the more widely distributed Atlantic (AT) lineage. Later mtDNA-RFLP (restriction fragment length polymorphism) studies detected the DU lineage in northern Spanish basins (Galicia). The aim of this work was to ascertain the origin and variability of these DU populations outside the Duero drainage. Using complete mtDNA control region sequencing, 11 novel DU haplotypes were identified. Several of them could be assigned to an endemic group in Galicia consistent with the long-time presence of the DU lineage outside the Duero River, and excluding a recent origin by human translocations. The DU haplotype group observed in north-western Iberian basins was estimated to diverge from that of the Duero River more than 100000 years ago. We therefore advocate for conservation strategies at regional and local scales rather than focussed in a single ESU as proposed in earlier works.

Development and characterization of novel microsatellite markers by Next Generation Sequencing for the blue and red shrimp Aristeus antennatus.

The blue and red shrimp, Aristeus antennatus, is a commercially important crustacean, in the Mediterranean Sea, which has been listed as a priority species for fishery management. Hypervariable microsatellite markers could be a useful tool to identify genetic stocks among geographically close fishing grounds. Potential microsatellite markers (97) identified from next-generation sequencing of an individual shrimp using a 454 GS Junior Pyrosequencer were tested on a preliminary panel of 15 individuals representing the four worldwide genetic stocks of the species from which 35 polymorphic loci were identified and used to characterize an additional 20 individuals from the Western Mediterranean Sea. In the Western Mediterranean sample, 32 out of 35 were polymorphic loci and the number of alleles per locus ranged from 2 to 14 and expected heterozygosity ranged from 0.050 to 0.968. No linkage disequilibrium was detected, indicating the independence of the loci. These novel microsatellites provide additional tools to address questions relating to genetic diversity, parentage studies and connectivity patterns of A. antennatus populations and help develop effective strategies to ensure long-term sustainability of this resource.