Mapping and functional characterisation of the Atlantic salmon genome and its regulation of pathogen response

Atlantic salmon is a species of both scientific and economic importance, and Atlantic salmon farming is a highly profitable industry worldwide. One of the biggest challenges being faced by farms, which affects production efficiency and results in severe economic loss, is disease. In livestock production, one of the approaches taken to limit the impact of disease outbreaks is to selectively breed for improved resistance within farmed populations. Although traditional family-based resistance breeding programs have shown improvements in resistance to a variety of bacterial, viral and parasitic diseases on Atlantic salmon farms, response to selection can be slow. One way of increasing selection efficiency is through the incorporation of genetic markers into breeding programs, for marker-assisted or genomic selection. However, genomic resources for cultured aquatic species are sparse, and the generation of new and denser resources for use in selective breeding programs would be advantageous. The main focus of this thesis is the development of genomic resources in Atlantic salmon and the application of those resources to gain a better understanding of the salmon genome, particularly in the genetic basis of host resistance to infectious diseases. The first aim of this thesis was to develop improved genomic resources for Atlantic salmon, and to characterise the Atlantic salmon genome via construction and analysis of a SNP linkage map derived from RAD-Sequencing (RAD-Seq). Approximately 6,500 SNPs were assigned to 29 linkage groups, and ~1,800 male-segregating, and ~1,400 female-segregating SNPs were ordered and positioned. Overall map lengths and recombination ratios were relatively consistent between the sexes and across the linkage groups (~1:1.5, male:female). However, a substantial difference in the degree of marker clustering was seen between males and females, which is reflective of the difference in the positions of chiasmata between the two sexes. Using this map, ~4,000 Atlantic salmon reference genome contigs were assigned to a linkage group, and 112 contigs were assigned to multiple linkage groups, highlighting regions of homeology (large sections of duplicated chromosomal regions) within the salmon genome. Alignment of SNP-flanking sequences to the stickleback and rainbow trout genomes identified putative gene-associated SNPs and cross-species chromosomal orthologies, and provided evidence in support of the salmonid-specific genome duplication. In addition, based on this and other publically available RAD-Seq datasets, the utility of RAD-Seq-derived data from different species and laboratories for population genetics analyses was tested. Short RAD-Seq contigs in Atlantic salmon and nine other teleost fish were used to identify cross-species orthologous genomic relationships. Several thousands of orthologous RAD loci were identified across the species, with the number of RAD loci decreasing with evolutionary distance, as expected. Previously published broad-level relationships between orthologous chromosomes were confirmed. The identified cross-species orthologous RAD loci were used to estimate evolutionary relationships between the ten teleost fish species. Previously published relationships were recovered, suggesting that RAD-Seq data derived from different laboratories is useful for this purpose. The second aim was to characterise the genetic architecture of resistance to two viral diseases affecting Atlantic salmon production on farms: pancreas disease (PD), and infectious pancreatic necrosis (IPN). Using data and samples collected from a large population of salmon fry challenged with PD, a high heritability for resistance was estimated (h2 ~0.5), and four QTL were identified, on chromosomes 3, 4, 7 and 23. The QTL explaining the highest within-family variation for resistance was located on chromosome 3. This QTL has been confirmed in a population of post-smolts by an independent research group, highlighting the potential for its incorporation into breeding programs to improve PD resistance. For IPN, the major resistance QTL had previously been mapped to linkage group 21. However, the mutation(s) underlying this QTL effect and the consequences of these mutation(s) on the affected genes and relevant biological resistance mechanisms are unknown. To generate a list of candidate genes within the vicinity of the IPN QTL, QTL-linked DNA sequences were aligned to four model fish genomes. This identified two QTL-orthologous regions in each of the species, and gene order within these regions was highly conserved across species. Analysis of gene expression patterns between IPN resistant and susceptible salmon in a viral challenge experiment revealed that the five most significantly differentially-expressed genes mapped to the QTL-orthologous region on linkage group II of stickleback. Pathway enrichment analysis across all differentially-expressed genes suggests that biological pathways influencing viral infection stress response/entry/replication, cellular energy production and apoptosis may be involved in resistance during the initial stages of IPN virus (IPNV) infection. These results have provided the basis for further study of the putative involvement of these candidate genes and pathways in genetic resistance to IPNV. In summary, the results and resources presented in this thesis extend our current understanding of the salmon genome and the genetic basis of resistance to two viral diseases, and provide resources with the potential to be used in Atlantic salmon selective breeding programs to tackle disease outbreaks.