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- Genetic implications of phylogeographical patterns in the conservation of the boreal wetland butterfly Colias palaeno (Pieridae)Publication . Kramp, Katja; Cizek, Oldrich; Madeira, Pedro M.; Ramos, Ana; Konvicka, Martin; Castilho, Rita; Schmitt, ThomasThe boreo-montane wetland butterfly species Colias palaeno has a European distribution from the Alps to northern Fennoscandia. Within its European range, the species' populations have shrunk dramatically in recent historical times. Therefore, detailed baseline knowledge of the genetic makeup of the species is pivotal in planning potential conservation strategies. We collected 523 individuals from 21 populations across the entire European range and analyzed nuclear (20 allozyme loci) and mitochondrial (600bp of the cytochrome c oxidase subunit I gene) genetic markers. The markers revealed contrasting levels of genetic diversity and divergence: higher in allozymes and lower in mitochondrial sequences. Five main groups were identified by allozymes: Alps, two Czech groups, Baltic countries, Fennoscandia, and Poland. The haplotype mitochondrial network indicates a recent range expansion. The most parsimonious interpretation for our results is the existence of a continuous Wurm glacial distribution in Central Europe, with secondary disjunction during the Last Glacial Maximum into a south-western and a north-eastern fragment and subsequent moderate differentiation. Both groups present signs of postglacial intermixing in the Czech Republic. However, even a complete extinction in this region would not considerably affect the species' genetic basis, as long as the source populations in the Alps and in northern Europe, comprising the most relevant evolutionary units for conservation, are surviving.
- Distinctive genetic signatures of two fairy shrimp species with overlapping ranges in Iberian temporary pondsPublication . Lopes Da Cunha, Regina; Sala, Jordi; Machado, Margarida; Boix, Dani; Madeira, Celine; Madeira, Pedro M.; Cristo, Margarida; Cancela Da Fonseca, Luís; Castilho, RitaTemporary lentic water bodies host biotic assemblages adapted to the transient nature of these freshwater habitats. Fairy shrimps (Crustacea, Branchiopoda, Anostraca) are one of the most important biological components of these unique environments and have a fossil record dating back to the Middle Jurassic (>150 million years). Some anostracan species show a geographically restricted distribution, whereas others are widely dispersed. We aimed to investigate the relationship between different geographic extents and patterns of genetic structure in species of Anostraca. Following this objective, we selected two species with contrasting ranges but overlapping geographic distributions and similar life-history traits in the study area. We analysed additional information that, from an ecological (e.g. egg-bank, niche breadth, and pond connectivity) and evolutionary (e.g. crown-group age of each species) perspective, may explain the obtained phylogeographic patterns. Between 2005 and 2018, we sampled two species of fairy shrimps (309 specimens of Branchipus cortesi and 264 specimens of Tanymastix stagnalis) from 53 temporary ponds of Portugal. We added five other locations from Spain and France to include other European locations for T. stagnalis. Additionally, we also sampled Branchipus schaefferi from two temporary water bodies (Spain and Morocco) to include in the dating analysis. Reconstructed phylogenies based on mitochondrial sequence data indicate the existence of deeply divergent clades with an unequivocal phylogeographic structure in T. stagnalis and shallower divergences in B. cortesi with a less clear geographic correspondence. We found evidence of frequent local and rare long-distance dispersal events in both species and limited intermediate dispersal, which was more common in B. cortesi. A Bayesian dating analysis using the Branchiopoda fossil record estimated the age of the most recent common ancestors of T. stagnalis and B. cortesi at 32.4 and 12.8 million years, respectively. Haplotype accumulation curves indicated that only a portion of the genetic composition of the species was sampled on each hydroperiod and showed the existence of large, genetically diverse egg banks that remain in the soil. These egg banks represent a genetic reservoir that guarantees the survival of the species because active populations from different hydroperiods may be genetically different and adapt to a changing environment. We hypothesise that the contrasting phylogeographic patterns displayed by the two fairy shrimp species may result from: (1) the earlier age of the most recent common ancestor of T. stagnalis, as older species have more time to accumulate mutations and, thus, are expected to exhibit higher genetic differentiation among populations; (2) slight differences in adult behaviour, life-history traits and cyst morphologies of T. stagnalis and B. cortesi favouring different animal dispersal vectors with distinct dispersal abilities. Therefore, phylogeographic patterns may be explained by both evolutionary and ecological processes, which operate in different time scales.
- Invasion genetics of the mummichog (Fundulus heteroclitus): recent anthropogenic introduction in IberiaPublication . Morim, Teófilo; Bigg, Grant R.; Madeira, Pedro M.; Palma, Jorge; Duvernell, David D.; Gisbert, Enric; Lopes Da Cunha, Regina; Castilho, RitaHuman activities such as trade and transport have increased considerably in the last decades, greatly facilitating the introduction and spread of non-native species at a global level. In the Iberian Peninsula, Fundulus heteroclitus, a small euryhaline coastal fish with short dispersal, was found for the first time in the mid-1970s. Since then, F. heteroclitus has undergone range expansions, colonizing the southern region of Portugal, southwestern coast of Spain and the Ebro Delta in the Mediterranean Sea. Cytochrome b sequences were used to elucidate the species invasion pathway in Iberia. Three Iberian locations (Faro, Cádiz and Ebro Delta) and 13 other locations along the native range of F. heteroclitus in North America were sampled. Results revealed a single haplotype, common to all invasive populations, which can be traced to the northern region of the species' native range. We posit that the origin of the founder individuals is between New York and Nova Scotia. Additionally, the lack of genetic structure within Iberia is consistent with a recent invasion scenario and a strong founder effect. We suggest the most probable introduction vector is associated with the aquarium trade. We further discuss the hypothesis of a second human-mediated introduction responsible for the establishment of individuals in the Ebro Delta supported by the absence of adequate muddy habitats linking Cádiz and the Ebro Delta. Although the species has a high tolerance to salinity and temperature, ecological niche modelling indicates that benthic habitat constraints prevent along-shore colonisation suggesting that such expansions would need to be aided by human release.
- Sex-dependent and-independent transcriptional changes during haploid phase gametogenesis in the sugar kelp Saccharina latissimaPublication . Pearson, Gareth; Martins, Neusa; Madeira, Pedro M.; Serrao, Ester; Bartsch, InkaIn haplodiplontic lineages, sexual reproduction occurs in haploid parents without meiosis. Although widespread in multicellular lineages such as brown algae (Phaeophyceae), haplodiplontic gametogenesis has been little studied at the molecular level. We addressed this by generating an annotated reference transcriptome for the gametophytic phase of the sugar kelp, Saccharina latissima. Transcriptional profiles of microscopic male and female gametophytes were analysed at four time points during the transition from vegetative growth to gametogenesis. Gametogenic signals resulting from a switch in culture irradiance from red to white light activated a core set of genes in a sex-independent manner, involving rapid activation of ribosome biogenesis, transcription and translation related pathways, with several acting at the post-transcriptional or post-translational level. Additional genes regulating nutrient acquisition and key carbohydrate-energy pathways were also identified. Candidate sex-biased genes under gametogenic conditions had potentially key roles in controlling female- and male-specific gametogenesis. Among these were several sex-biased or -specific E3 ubiquitin-protein ligases that may have important regulatory roles. Females specifically expressed several genes that coordinate gene expression and/or protein degradation, and the synthesis of inositol-containing compounds. Other female-biased genes supported parallels with oogenesis in divergent multicellular lineages, in particular reactive oxygen signalling via an NADPH-oxidase. Males specifically expressed the hypothesised brown algal sex-determining factor. Male-biased expression mainly involved upregulation of genes that control mitotic cell proliferation and spermatogenesis in other systems, as well as multiple flagella-related genes. Our data and results enhance genome-level understanding of gametogenesis in this ecologically and economically important multicellular lineage.
- Cryptic diversity in southern African kelpPublication . Madeira, Pedro; Reddy, Maggie M.; Assis, Jorge; Bolton, John J.; Rothman, Mark D.; Anderson, Robert J.; Kandjengo, Lineekela; Kreiner, Anja; Coleman, Melinda A.; Wernbergh, Tomas; De Clerck, Olivier; Leliaert, Frederik; Bandeira, Salomão; Ada, Abdul M.; Miranda Neiva, João; Pearson, Gareth Anthony; Serrao, Ester A.The southern coast of Africa is one of the few places in the world where water temperatures are predicted to cool in the future. This endemism-rich coastline is home to two sister species of kelps of the genus Ecklonia maxima and Ecklonia radiata, each associated with specific thermal niches, and occuring primarily on opposite sides of the southern tip of Africa. Historical distribution records indicate that E. maxima has recently shifted its distribution similar to 70 km eastward, to sites where only E. radiata was previously reported. The contact of sister species with contrasting thermal affinities and the occurrence of mixed morphologies raised the hypothesis that hybridization might be occurring in this contact zone. Here we describe the genetic structure of the genus Ecklonia along the southern coast of Africa and investigate potential hybridization and cryptic diversity using a combination of nuclear microsatellites and mitochondrial markers. We found that both species have geographically discrete genetic clusters, consistent with expected phylogeographic breaks along this coastline. In addition, depth-isolated populations were found to harbor unique genetic diversity, including a third Ecklonia lineage. Mito-nuclear discordance and high genetic divergence in the contact zones suggest multiple hybridization events between Ecklonia species. Discordance between morphological and molecular identification suggests the potential influence of abiotic factors leading to convergent phenotypes in the contact zones. Our results highlight an example of cryptic diversity and hybridization driven by contact between two closely related keystone species with contrasting thermal affinities.