Fish diversification at the pace of geomorphological changes: evolutionary history of western Iberian Leuciscinae (Teleostei: Leuciscidae) inferred from multilocus sequence data.

The evolutionary history of western Iberian Leuciscinae, obligatory freshwater fish, is directly linked to the evolution of the hydrographic network of the Iberian Peninsula after its isolation from the rest of Europe, which involved dramatic rearrangements such as the transition from endorheic lakes to open basins draining to the Atlantic. Previous phylogenetic research on western Iberian leuciscines, using mainly mitochondrial DNA and more recently one or two nuclear genes, has found contradictory results and there remain many unresolved issues regarding species relationships, taxonomy, and evolutionary history. Moreover, there is a lack of integration between phylogenetic and divergence time estimates and information on the timing of geomorphological changes and paleobasin rearrangements in the Iberian Peninsula. This study presents the first comprehensive fossil-calibrated multilocus coalescent species tree of western Iberian Leuciscinae (including 14 species of Achondrostoma, Iberochondrostoma, Pseudochondrostoma and Squalius endemic to the Iberian Peninsula, seven of which endemic to Portugal) based on seven nuclear genes, and from which we infer their biogeographic history by comparing divergence time estimates to known dated geological events. The phylogenetic pattern suggests slow-paced evolution of leuciscines during the Early-Middle Miocene endorheic phase of the main Iberian river basins, with the shift to exorheism in the late Neogene-Quaternary allowing westward dispersals that resulted in many cladogenetic events and a high rate of endemism in western Iberia. The results of this study also: (i) confirm the paraphyly of S. pyrenaicus with respect to S. carolitertii, and thus the possible presence of a new taxon in the Portuguese Tagus currently assigned to S. pyrenaicus; (ii) support the taxonomic separation of the Guadiana and Sado populations of S. pyrenaicus; (iii) show the need for further population sampling and taxonomic research to clarify the phylogenetic status of A. arcasii from the Minho basin and of the I. lusitanicum populations in the Sado and Tagus basins; and (iv) indicate that A. occidentale, I. olisiponensis and P. duriensis are the most ancient lineages within their respective genera.

Whole-plant and leaf determinants of growth rates in progenies of Genipa americana L. (Rubiaceae).

Genipa americana (Rubiaceae) is a fruit tree with broad phytogeographic domain and suitable for different silvicultural systems in the tropics. The knowledge associated with the relative growth rate of species such as G. americana, provides important guidelines for the effective establishment and survival of seedlings after planting in the field. In this study we investigated differences in growth, biomass allocation and photosynthesis of seedlings originating from different mother plants of G. americana in southern Bahia, Brazil. For this, we evaluated fifteen variables associated with carbon balance at the whole plant and leaf scales of twelve G. americana progenies. All seedlings grew over a period of 198 days under similar microclimatic conditions with approximately 65% full sun. Our results showed significant differences in the relative growth rates (RGR), with the highest and lowest mean values being 29.0 and 38.0 mg g-1 day-1, respectively. Differences in RGR between G. americana progenies were highly related to differences in biomass allocation at both whole plant and leaf scales. From a practical point of view, we demonstrate that the selection of mother plants to produce seedlings with higher growth rates, and consequently greater establishment capacity in field plantings, can be made from evaluations of growth and biomass allocation variables at the whole plant scale.

Applying genomic approaches to delineate conservation strategies using the freshwater mussel Margaritifera margaritifera in the Iberian Peninsula as a model.

Effective conservation actions to counteract the current decline of populations and species require a deep knowledge on their genetic structure. We used Single Nucleotide Polymorphisms (SNPs) to infer the population structure of the highly threatened freshwater pearl mussel Margaritifera margaritifera in the Iberian Peninsula. A total of 130 individuals were collected from 26 locations belonging to 16 basins. We obtained 31,692 SNPs through Genotyping by Sequencing (GBS) and used this dataset to infer population structure. Genetic diversity given as observed heterozygosity was low. Pairwise FST comparisons revealed low levels of genetic differentiation among geographically close populations. Up to 3 major genetic lineages were determined: Atlantic, Cantabrian and Douro. This structure suggests a close co-evolutionary process with brown trout (Salmo trutta), the primordial fish host of this mussel in the studied area. Some sub-basins showed some genetic structuring, whereas in others no intrapopulation differentiation was found. Our results confirm that genetic conservation units do not match individual basins, and that knowledge about the genetic structure is necessary before planning recovery plans that may involve relocation or restocking. The same reasoning should be applied to strictly freshwater species that are sessile or have restricted dispersal abilities and are currently imperiled worldwide.

Comparisons of the genetic structure of Squalius populations (Teleostei, Cyprinidae) from rivers with contrasting histories, drainage areas and climatic conditions based on two molecular markers.

The genetic structure of Squalius populations from Portuguese Atlantic- and Mediterranean-type streams (assigned to six distinct morphoclimatic regions) was compared using sequences of the cytb and beta-actin genes. The drainage area was significantly correlated to all the genetic diversity indices, pointing to the need to control for this effect in comparing populations with different histories and paleoecologies. A significant correlation was also found between genetic diversity and the morphoclimatic regions to which the rivers were assigned, with the highest diversity in warmer, lower latitude, Mediterranean-type streams. This relationship was not due to idiosyncratic characteristics of the clades compared. When the drainage area and phylogenetic effects were removed, the southern Mediterranean streams harboured significantly more genetic diversity even when compared with much larger northern streams. It is argued that these results are likely caused both by the metapopulation structure of the Mediterranean streams and by the severe reduction or local extinction of populations in the northern rivers during glaciations.

Reading the history of a hybrid fish complex from its molecular record.

Squalius alburnoides is a widely distributed intergeneric hybrid complex with fish of both sexes, varying ploidy levels and proportions of the parental genomes. Its dispersal routes were here delineated and framed by the reconstruction of the phylogeny and phylogeography of other Squalius with which it hybridizes, based on the available data on the paleohydrographical history of the Iberian Peninsula. Results based on sequences of cytochrome b and beta-actin genes showed that: proto-Squalius pyrenaicus originated at least five species as it dispersed throughout the Iberian Peninsula in the Mio-Pliocene; the S. alburnoides complex likely had a single origin in the bulk of Iberia, in the Upper Tagus/Guadiana area, when hydrographical rearrangements allowed the contact between its ancestors (around 700,000 years ago); interspecific crosses allowed the introgression of mitochondrial and nuclear genes of S. alburnoides in allopatric species/populations of other Squalius and vice-versa; and reconstituted S. alburnoides non-hybrid males may contribute to the replacement of the typical mtDNA of the complex (in the populations where they occur, crosses with females of other Squalius seem to have been especially frequent). A number of dispersal events and colonization routes are proposed.