Paulo Alexandrino

Mitochondrial haplotype diversity among Portuguese brown trout Salmo trutta L. populations: relevance to the post-Pleistocene recolonization of northern Europe

Mitochondrial haplotype diversity in seven Portuguese populations of brown trout, Salmo trutta L., was investigated by sequencing the 5′ end of the mitochondrial DNA (mtDNA) control region. Five new haplotypes were described for this species, each two to three mutational steps distant from the common north Atlantic haplotype. Significant population subdivision of mtDNA haplotypes was also apparent. Based on these results, as well as on published data describing the distribution of both mtDNA haplotypes and allozyme alleles throughout Europe, the postglacial recolonization of northern Europe was re‐evaluated. It is argued that the available data do not support the contribution of two major glacial refugia (southwest Atlantic and Ponto‐Caspian Basin) to this postglacial recolonization, as proposed in a recently published model. The unique genetic architecture of Portuguese brown trout within the Atlantic‐basin clade of this species represents a highly valuable genetic resource that should be protected from introgression with nonendemic strains of hatchery fish.

Patterns of colonization, evolution and gene flow in species of the genus Patella in the Macaronesian Islands.

The study of phylogeographical patterns may contribute to a better understanding of factors affecting the dispersal of organisms in ecological and historical times. For intertidal organisms, islands are particularly suitable models allowing the test of predictions related to the efficacy of pelagic larvae dispersal. Here, we study the phylogeographical patterns and gene flow within three groups of species of the genus Patella present in the Macaronesian Islands that have been previously shown to be monophyletic. The genetic variability of around 600 bp of the mitochondrial gene cytochrome c oxidase subunit I was studied by single strand conformation polymorphism and/or sequencing for seven species of limpets. A total of 420 samples were analysed from the Macaronesian archipelagos, North Africa, and Atlantic and Mediterranean shores of the Iberian Peninsula. No clear geographical pattern or temporal congruence was found between the three groups of species, pointing to independent histories and colonization events. However, for the three groups, the split between the Macaronesian and the mainland forms most probably occurred before 3.9 million years ago, predating the establishment of the current circulation patterns. The presence of pelagic larvae in these species is shown to be insufficient to ensure gene flow between continental and Macaronesian populations and between the Macaronesian archipelagos. In the endangered Azorean populations of Patella candei, there is restricted gene flow to Flores and Graciosa.

Barriers to gene flow in the marine environment: insights from two common intertidal limpet species of the Atlantic and Mediterranean.

Knowledge of the scale of dispersal and the mechanisms governing gene flow in marine environments remains fragmentary despite being essential for understanding evolution of marine biota and to design management plans. We use the limpets Patella ulyssiponensis and Patella rustica as models for identifying factors affecting gene flow in marine organisms across the North-East Atlantic and the Mediterranean Sea. A set of allozyme loci and a fragment of the mitochondrial gene cytochrome C oxidase subunit I were screened for genetic variation through starch gel electrophoresis and DNA sequencing, respectively. An approach combining clustering algorithms with clinal analyses was used to test for the existence of barriers to gene flow and estimate their geographic location and abruptness. Sharp breaks in the genetic composition of individuals were observed in the transitions between the Atlantic and the Mediterranean and across southern Italian shores. An additional break within the Atlantic cluster separates samples from the Alboran Sea and Atlantic African shores from those of the Iberian Atlantic shores. The geographic congruence of the genetic breaks detected in these two limpet species strongly supports the existence of transpecific barriers to gene flow in the Mediterranean Sea and Northeastern Atlantic. This leads to testable hypotheses regarding factors restricting gene flow across the study area.

Comparative phylogeography and demographic history of European shads (Alosa alosa and A. fallax) inferred from mitochondrial DNA

BackgroundComparative broad-scale phylogeographic studies of aquatic organisms provide insights on biotic responses to the paleohydrological dynamics associated with climatic oscillations. These insights can be used to formulate a framework for understanding the evolutionary history of a species or closely related taxa as well as aid in predictive modeling of further responses to climate change. Anadromous fishes constitute interesting models for understanding the relative importance of environmental versus biological factors in shaping intraspecific genetic substructure on the interface between marine and freshwater realms. European shads, Alosa alosa and A. fallax are anadromous species that have persisted through historical large-scale environmental perturbations and now additionally face an array of anthropogenic challenges. A comprehensive phylogeographic investigation of these species is needed to provide insights on both the historical processes that have shaped their extant genetic structure and diversity, and the prospects for their future management and conservation.ResultsDespite introgressive hybridization, A. alosa and A. fallax are genetically divergent, congruent with previous studies. Three similarly divergent mtDNA clades were recognized within both A. fallax and A. alosa, most likely originating during common periods of isolation during the Pleistocene among the studied oceanographic regions. Periods of basin isolation apparently extended to the Black Sea as additional Alosa clades occur there. The present day geographic distribution of genetic diversity within European Alosa sp. suggests the existence of a strong but permeable barrier between the Atlantic and Mediterranean seas, as shown for a number of other aquatic species. Overall mtDNA diversity is considerably lower for A. alosa compared to A. fallax, suggesting that the former species is more sensitive to climatic as well as anthropogenic changes. For A. fallax, migration from the Mediterranean to the Atlantic was detected but not in the opposite direction, with colonization of the North Atlantic probably occurring after last glacial maximum.ConclusionThe similar haplotype network topologies between the two species support a common intraspecific history of isolation. Despite these similarities, A. alosa and A. fallax have clearly responded differently to the hydrological dynamics of the Pleistocene, as reflected in their distinct demographic histories. As the species additionally occupy different ecological niches it should not be surprising that they differ in resilience to natural or human-mediated climatic changes. For A. fallax, it is further clear that its demographic response to large-scale hydrological events is not synchronized between the Atlantic and Mediterranean basins. These regional and species-specific differences should be incorporated into future predictive modeling of biological response to climate change as well as current management concepts.

Phylogeographical history of the white seabream Diplodus sargus (Sparidae): Implications for insularity

Abstract Partial sequences of the mitochondrial control region and its comparison with previously published cytochrome b (cyt-b) and microsatellite data were used to investigate the influence of island isolation and connectivity on white seabream genetic structure. To achieve this, a total of 188 individuals from four island localities (Castellamare and Mallorca, Mediterranean Sea; Azores and Canary Islands, Atlantic Ocean) and five coastal localities (Banyuls, Murcia and Tunisia, Mediterranean Sea; Galicia and Faro, Atlantic Ocean) were analysed. Results showed high haplotype diversity and low to moderate nucleotide diversity in all populations (except for the Canary Islands). This pattern of genetic diversity is attributed to a recent population expansion which is corroborated by other results such as cyt-b network and demographic analyses. Low differentiation among Mediterranean/Atlantic and coastal/island groups was shown by the AMOVA and FST values, although a weak phylogeographic break was detected using cyt-b data. However, we found a clear and significant island/distance effect with regard to the Azores islands. Significant genetic differentiation has been detected between the Azores islands and all other populations. The large geographical distance between the European continental slope and the Azores islands is a barrier to gene flow within this region and historic events such as glaciation could also explain this genetic differentiation.

The evolutionary history of sharp- and blunt-snouted lenok (Brachymystax lenok (Pallas, 1773)) and its implications for the paleo-hydrological history of Siberia.

BackgroundBroad-scale phylogeographic studies of freshwater organisms provide not only an invaluable framework for understanding the evolutionary history of species, but also a genetic imprint of the paleo-hydrological dynamics stemming from climatic change. Few such studies have been carried out in Siberia, a vast region over which the extent of Pleistocene glaciation is still disputed. Brachymystax lenok is a salmonid fish distributed throughout Siberia, exhibiting two forms hypothesized to have undergone extensive range expansion, genetic exchange, and multiple speciation. A comprehensive phylogeographic investigation should clarify these hypotheses as well as provide insights on Siberias paleo-hydrological stability.ResultsMolecular-sequence (mtDNA) based phylogenetic and morphological analysis of Brachymystax throughout Siberia support that sharp- and blunt-snouted lenok are independent evolutionary lineages, with the majority of their variation distributed among major river basins. Their evolutionary independence was further supported through the analysis of 11 microsatellite loci in three areas of sympatry, which revealed little to no evidence of introgression. Phylogeographic structure reflects climatic limitations, especially for blunt-snouted lenok above 56° N during one or more glacial maxima. Presumed glacial refugia as well as interbasin exchange were not congruent for the two lineages, perhaps reflecting differing dispersal abilities and response to climatic change. Inferred demographic expansions were dated earlier than the Last Glacial Maximum (LGM). Evidence for repeated trans-basin exchange was especially clear between the Amur and Lena catchments. Divergence of sharp-snouted lenok in the Selenga-Baikal catchment may correspond to the isolation of Lake Baikal in the mid-Pleistocene, while older isolation events are apparent for blunt-snouted lenok in the extreme east and sharp-snouted lenok in the extreme west of their respective distributions.ConclusionSharp- and blunt-snouted lenok have apparently undergone a long, independent, and demographically dynamic evolutionary history in Siberia, supporting their recognition as two good biological species. Considering the timing and extent of expansions and trans-basin dispersal, it is doubtful that these historical dynamics could have been generated without major rearrangements in the paleo-hydrological network, stemming from the formation and melting of large-scale glacial complexes much older than the LGM.

Development of microsatellite loci for the black-footed limpet, Patella depressa , and cross-amplification in two other Patella species

Limpets (Patella spp.) are suitable organisms to investigate the effects of climate change in marine systems. They are widespread over NE Atlantic intertidal rocky shores and have been extensively studied in terms of population dynamics and ecology. Within genus Patella, microsatellites have only been developed for Patella caerulea and cross-species tests are unknown. In this work, we describe 11 primer pairs for Patella depressa and the results of cross-species testing on Patella candei and Patella rustica.

Evidence for genetic differentiation in the European conger eel Conger conger based on mitochondrial DNA analysis

The European conger eel Conger conger is an important marine benthic fish in the North-East Atlantic and represents a valuable fishery resource. However, little is known about its reproductive biology. In an attempt to gain a better understanding of the conger eel population structure, mitochondrial DNA (mtDNA) sequences were examined. A region with 432 bp of the control region of the mtDNA was sequenced from 40 individuals from six different locations around the central and eastern North Atlantic Ocean. Thirty variable positions defined 28 distinct haplotypes. The average sequence difference within samples (1.3–4.2%) was comparable to those between samples (1.4–3.6%). MtDNA sequence-based statistical tests showed significant geographic differentiation between some local population samples, suggesting that the conger eel does not comprise a single panmictic population. However, given our sample sizes, these preliminary results should be interpreted with caution and more individuals from more sites, including the Mediterranean Sea, should be analyzed in detail. The genetic variability detected in this study is an initial step to elucidate the genetic back-ground of the conger eel population structure.

A highly polymorphic plasma protein locus in brown trout (Salmo trutta L.) populations from Portugal.

Genetic polymorphism of an unidentified plasma protein (PX) is described for the first time in Salmo trutta (L.) by means of isoelectric focusing. The analysis of 414 individuals from different geographic origins in Portugal allowed the identification of nine alleles. Heterozygosity in natural populations is generally above 0.60, thus giving similar values to those reported for brown trout microsatellite loci. Substructuring of Portuguese brown trout is evident between northern and southern basins. Genetic affinities between the southernmost rivers and the hatchery stock were detected, suggesting the existence of recent stocking influences.

PANMIXIA IN THE ENDANGERED SLIPPER LOBSTER SCYLLARIDES LATUS FROM THE NORTHEASTERN ATLANTIC AND WESTERN MEDITERRANEAN

In the marine realm, it is important to understand the connectivity and population dynamics of commercially exploited species; this information promotes sustainable fishing practices. The large slipper lobster, Scyllarides latus (Latreille, 1803), is a commercially valuable crustacean that has been over-exploited in many areas throughout its distribution range in the northeastern (hereafter NE) Atlantic and the Mediterranean. Here, we report the first molecular population genetic study of S. latus based on 9 nuclear microsatellite loci. A total of 128 specimens of S. latus were collected in the Western Mediterranean (2 locations) and four regions (13 locations) in the NE Atlantic, including Southern Portugal and the Macaronesian archipelagos of Azores, Canary Islands and Cape Verde. Maximum number of alleles per locus ranged from 4 to 14, and observed heterozygosity per locus ranged from 0.143 to 0.795 (0.539 ± 0.087, mean ± SE). Pairwise region estimates of FST, RST and Dest were very low (<0.040) and not significant in all comparisons. Bayesian clustering analysis also suggested homogeneity in S. latus across all regions. The Almeria-Oran front, an important biogeographic boundary between the Atlantic and Mediterranean basins, was not found to have a significant impact on the genetic structuring of S. latus. Overall, all of our analyses of genetic differentiation and migration suggested panmixia in S. latus across its distributional range. Such pattern is likely to result from its high fecundity and long-lived pelagic larva, which can promote high levels of connectivity between geographically distant populations. Future conservation strategies should manage all S. latus populations jointly (one stock).