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Bargelloni, L.

Publications and source records attributed to Bargelloni, L..

9 recordsLinked to original sources

The epigenomic landscape of deep lineage divergence: The case of the European sea bass

BackgroundUnderstanding the role of non-coding genomic variation in speciation remains a major challenge in evolutionary biology. Here, we investigated whether regulatory elements contribute to this process between Atlantic and Mediterranean lineages of European sea bass (Dicentrarchus labrax), a well-characterized case-study near speciation where barriers to introgression exist in the presence of connectivity between diverging populations. ResultsWe generated a novel, highly contiguous genome assembly, which was annotated at the epigenomic level using ATAC-seq and ChIP-seq with six embryonic developmental stages and five tissue types in adult fish, identifying thousands of promoters, enhancers, and open chromatin regions. Integrating this annotation with whole-genome sequence data from 65 individuals across three geographically distinct populations, we identified 57,505 outlier SNPs and 332 structural variants (SVs) showing elevated differentiation between Atlantic and East Mediterranean lineages. Outlier SVs affected key regulatory elements and coding genes, while outlier SNPs were enriched in regulatory elements, particularly enhancers active in adult tissues. Local genomic divergence correlated positively with regulatory element density, especially on chromosomes 1, 9, and 18, which are enriched in genes related to osmoregulation, immune response, and oxidative stress -- processes relevant to adaptation across contrasting marine environments. ConclusionsThese findings support a major role for regulatory variation in driving deep lineage divergence through local adaptation.

evolutionary biology↗

MicroRNA modulation of viral nervous necrosis resistance in European seabass

MicroRNAs (miRNAs) are key post-transcriptional regulators of antiviral immunity, controlling gene expression by targeting 3 UTRs of immune-related transcripts. Despite their importance, the role of miRNAs in viral nervous necrosis (VNN) resistance in European seabass (Dicentrarchus labrax) is unexplored. Here, we characterized for the first time the brain miRNome of seabass from three VNN-resistance genotypes (susceptible, intermediate, resistant) across two genetically distinct seabass clusters. Differential expression analyses revealed cluster-specific patterns, with susceptible fish consistently showing overexpression of the differently expressed miRNAs (DEmiRNAs) as compared to the resistant fish. Considering the two genetic clusters in the study, miR-199-5p was differentially expressed between the VNN susceptible and resistant fish. This miRNA was found to be less expressed in the resistant individuals. Functional characterization of the miRNA predicted that it binds to two distinct miRNA recognition elements (MREs) within the ifi27l2a 3 UTR. These MREs flank a SNP (Chr3:10,082,380) previously associated with VNN survival. A strong negative correlation (r= -0.840) between miR-199-5p expression and ifi27l2a mRNA abundance further supports a post-transcriptional repression mechanism. Together, these results propose a regulatory model in which miR-199-5p modulates ifi27l2a expression, contributing to phenotypic variation in VNN resistance and positioning it as a promising biomarker for seabass aquaculture breeding.

genomics↗

A dual-clam species 63K SNP array for sustainable production and conservation of wild resources

Bivalves play an essential role in coastal ecosystems and their aquaculture represents an important economic sector in Europe playing a pivotal role within the EU Blue Growth Strategy. Among clam species, the Manila clam, Ruditapes philippinarum, and the grooved carpet shell, R. decussatus, are within the top five species in terms of production volume and economic value. In this study we designed and validated the first medium-density 63K single nucleotide polymorphism (SNP) array for these two commercially important species. By leveraging a new chromosome-level genome assembly for R. philippinarum and that of R. decussatus, we identified over 300 million SNPs through whole-genome resequencing and genotyping-by-sequencing strategies. After stringent filtering, we selected 49,392 high-quality SNPs for R. philippinarum and 14,193 for R. decussatus to construct a dual-species array. Array validation was carried out by genotyping 384 individuals across multiple wild populations and hatchery samples, demonstrating excellent performance, with 67.7% and 67.5% of SNPs classified as high-quality polymorphic markers for R. philippinarum and R. decussatus, respectively. Minor allele frequency, missing data rate, and inter-marker distance met stringent quality thresholds, confirming the array robustness for clam population genetics. Parentage analysis in R. philippinarum families highlighted significant power for pedigree reconstruction in breeding programs. This publicly available genomic resource provides a reliable, cost-effective genotyping platform to enable population genomics and advanced selective breeding, genome-wide association studies, and genetic monitoring, ultimately strengthening management of genetic diversity and sustainable farming for two key clam species.

genomics↗

Comparative transcriptomics of immune response to viral and bacterial stimuli in three acanthopterygian bony fish

Diseases triggered by bacterial and viral infections have caused huge economic losses for three of the most important European aquaculture species: turbot (Scophthalmus maximus), gilthead seabream (Sparus aurata) and European seabass (Dicentrarchus labrax). Understanding how they respond to pathogens is relevant for advancing aquaculture disease management and comprehending evolution of immune response within teleosts. Since mechanisms conserved across species are assumed to perform important roles, comparative analysis provides a powerful approach to pinpoint key elements of the immune defence. Here, we report the first comparative immune-transcriptomic analysis of these three species using bacterial and viral mimics after 20-24 hours post-stimulation with inactivated Vibrio anguillarum and Poly I:C in the head kidney of live fish (in vivo), and in primary leukocyte cultures (in vitro). The transcriptomic response, based on RNA-seq data, revealed a total of 503 differentially expressed orthologous genes in response to in vitro-Poly I:C, 1,472 to in vitro-Vibrio, 920 to in vivo-Poly I:C, and 832 to in vivo-Vibrio. Interestingly, consistent expression patterns were identified in seven genes across all species in both cell culture and live organisms in response to both pathogen stimuli. Functional enrichment analysis revealed associations with immunity, DNA replication and repair, and cytokine pathways, with the Toll-Like Receptor (TLR) pathway common to both conditions and stimuli. Our study suggests conservation of orthologous gene expression during infection across the three species for genes involved in chemokine pathways, interferon signalling, antigen processing and presentation, cell signalling regulators, and MAPK cascades. This study provides insights into key immune defence mechanisms in acanthopterygian bony fish.

genomics↗

Integrated functional genomic analysis identifies the regulatory variants underlying a major QTL for disease resistance in European sea bass

BackgroundViral nervous necrosis (VNN) is a viral disease threatening the sustainability of global aquaculture, and affecting over 50 of farmed and ecologically important species. A major QTL for resistance to VNN has been previously described in European sea bass, but the underlying causal gene(s) and mutation(s) are unknown. To identify the mechanisms and genetic factors underpinning resistance to VNN, we integrated farmed and wild genetic data with multiple functional genomics assays in a farmed European sea bass population. ResultsA high heritability (h2 [~] 0.40) was estimated for VNN resistance. A major QTL for this trait was confirmed on chromosome 3, and whole-genome resequencing narrowed its location to a small region containing 4 copies of interferon alpha inducible protein 27-like 2A (IFI27L2A) genes, and one copy of the interferon alpha inducible protein 27-like 2 (IFI27L2) gene. RNA sequencing revealed a clear association between the QTL genotype and the expression of two of the IFI27L2A genes, and the IFI27L2 gene. Integration with chromatin accessibility and histone modification data pinpointed two SNPs in active regulatory regions of two of these genes (IFI27L2A and IFI27L2), and transcription factor binding site gains for the resistant alleles were predicted. These alleles, particularly the SNP variant CHR3:10077301, exhibited higher frequency in Eastern Mediterranean sea bass populations, which show considerably higher levels of resistance to VNN. ConclusionsThe SNP variant CHR3:10077301, through modulation of IFI27L2 and IFI27L2A genes, is likely the causative mutation underlying resistance to VNN in European sea bass. This is one of the first causative mutations discovered for disease resistance traits, and paves the way for marker-assisted selection as well as biotechnological approaches to enhance resistance to VNN in European sea bass and other susceptible species.

genetics↗

Generating gnotobiotic bivalves: a new method on Manila clam (Ruditapes philippinarum)

The microbiome, which encompasses microbial communities associated with animal hosts, exerts a profound impact on host physiology and ecosystem dynamics. The application of advanced sequencing technologies has enabled researchers to investigate the composition of microbiomes across a range of hosts and environments. While correlating microbial composition with health outcomes has been a priority, interpreting such data requires caution to avoid overemphasizing the roles of microbes. Understanding microbial influence demands mechanistic insights, which are often elucidated through gnotobiology. Despite their limitations in representing animal diversity, model organisms offer the advantage of reproducibility and experimental tractability. However, the marine realm, especially bivalves, which are crucial for ecosystem functioning and aquaculture, lacks gnotobiotic models. In this study, we present a method for generating microbiome-depleted and gnotobiotic clams (Ruditapes philippinarum), one of the most widely farmed molluscs in the world and a sentinel organism for climate change. This model expands gnotobiotic research into marine invertebrates, thereby enabling investigations into the impact of microbes on such key animal species.

microbiology↗

Transcriptional profiling of populations in the clam Ruditapes decussatus suggests genetically determined differentiation in gene-expression along parallel temperature gradients and between races of the Atlantic Ocean and West Mediterranean sea

Ongoing ocean warming due to climate change poses new challenges for marine life and its exploitation. We have used transcriptomics to find genetically-based responses to increased temperature in natural populations of the marine clam Ruditapes decussatus living along parallel thermal gradients in southern Europe. Clams of the Atlantic and West Mediterranean races were collected in a northern (cool) and a southern (warm) localities. The animals were kept in running seawater in the warm, Atlantic southern locality for a 15-week period, during which water temperature raised to typical southern European summer values. After this period, the expression profile was obtained for a total of 34 clams and 11,025 probes by means of an oligonucleotide microarray. We found distinct transcriptional patterns for each population based on a total of 553 differentially expressed genes (DEG), indicating innate differences which probably have a genetic basis. Race and latitude contributed significantly to gene expression differences, with very different sets of DEG. A gene ontology analysis showed that races differed mainly at genes involved in ribosomal function and protein biosynthesis, while genes related to glutathione metabolism and ATP synthesis in the mitochondria were the most outstanding as to north/south transcriptional differences.

genomics↗

Development and validation of a combined species SNP array for the European seabass (Dicentrarchus labrax) and gilthead seabream (Sparus aurata)

SNP arrays are powerful tools for high-resolution studies of the genetic basis of complex traits, facilitating both population genomic and selective breeding research. The European seabass (Dicentrarchus labrax) and the gilthead seabream (Sparus aurata) are the two most important fish species for Mediterranean aquaculture. While selective breeding programmes increasingly underpin stocky supply for this industry, genomic selection is not yet widespread. Genomic selection has major potential to expedite genetic gain, in particular for traits practically impossible to measure on selection candidates, such as disease resistance and fillet yield. The aim of our study was to design a combined-species 60K SNP array for both European seabass and gilthead seabream, and to validate its performance on farmed and wild populations from numerous locations throughout the species range. To achieve this, high coverage Illumina whole genome sequencing of pooled samples was performed for 24 populations of European seabass and 27 populations of gilthead seabream. This resulted in a database of ~20 million SNPs per species, which were then filtered to identify high-quality variants and create the final set for the development of the MedFish SNP array. The array was then tested by genotyping a subset of the discovery populations and demonstrated a high conversion rate to functioning polymorphic assays on the array (92% in seabass: 89% in seabream) and repeatability (99.4 - 99.7%). The platform interrogates ~30K markers in each fish species, includes features such as SNPs previously shown to be associated with performance traits, and is enriched for SNPs predicted to alter protein function. The array was demonstrated to be effective at detecting population structure across a wide range of fish populations from diverse geographical origins, and to examine the extent of haplotype sharing among Mediterranean fish farms. Therefore, the MedFish array enables efficient and accurate high-throughput genotyping for genome-wide distributed SNPs on each fish species, and will facilitate stock management, population genomics approaches, and acceleration of selective breeding through genomic selection.

genomics↗

Genome-wide analysis clarifies the population genetic structure of wild Gilthead Sea Bream (Sparus aurata)

Gilthead sea bream is an important target for both recreational and commercial fishing in Europe, where it is also one of the most important cultured fish. Its distribution range goes from the Mediterranean to the African and European coasts of the North-East Atlantic. So far, the genetic structure of this species in the wild has been studied with microsatellite DNA, but the pattern of differentiation could not be fully clarified. In this study, almost 1000 wild sea bream from 23 locations in the Mediterranean Sea and Atlantic ocean where genotyped at 1159 SNP markers, of which 18 potentially under selection. Neutral markers suggested the presence of a weak subdivision into three genetic clusters: Atlantic, West and East Mediterranean. This last group could be further subdivided into an Ionian/Adriatic and an Aegean group using outlier markers. Seascape analysis suggested that this differentiation was mainly due to difference in salinity, and this was also supported by preliminary genomic functional analysis. These results are of fundamental importance for the development of proper management of this species in the wild and are a first step toward the study of the potential genetic impact of the sea bream aquaculture industry.View Full Text

ecology↗