Project description:Potential use of environmental DNA haplotyping for estimating the dispersal of an invasive fish, bluegill sunfish (Lepomis macrochirus), in Japan
Project description:Single-cell whole-genome haplotyping allows simultaneous detection of haplotypes associated with monogenic diseases, chromosome copy-numbering and subsequently, has revealed mosaicism in embryos and embryonic stem cells. Methods, such as karyomapping and haplarithmisis, were deployed as a generic and genome-wide approach for preimplantation genetic testing (PGT) and are replacing traditional PGT methods. While current methods primarily rely on SNP array, we envision sequencing-based methods to become more accessible and cost-efficient. Here, we developed a novel sequencing-based methodology to haplotype and copy-number profile single cells. Following DNA amplification, genomic size and complexity is reduced through restriction enzyme digestion and DNA is genotyped through sequencing. This single-cell genotyping-by-sequencing (scGBS) is the input for haplarithmisis, an algorithm we previously developed for SNP array-based single-cell haplotyping. We established technical parameters and developed an analysis pipeline enabling accurate concurrent haplotyping and copy-number profiling of single cells. We demonstrate its value in human blastomere and trophectoderm samples as application for PGT for monogenic disorders. Furthermore, we demonstrate the method to work in other species through analyzing blastomeres of bovine embryos. Our scGBS method opens up the path for single-cell haplotyping of any species with diploid genomes and could make its way into the clinic as a PGT application.
Project description:Streptococcal disease results in major mortality events of both marine and freshwater fishes worldwide. Streptococcus iniae is among the prominent causative bacterial strains as it has been found to cause a higher incidence of mortality and act as a zoonotic pathogen. Here we examine the susceptibility of two important aquaculture species in the United States, striped bass (Morone saxatilis) and white bass (Morone chrysops), to S. iniae. A high incidence of mortality was observed in both species, although striped bass succumbed more rapidly than white bass. Spleen gene expression at three time points following infection was analyzed to further elucidate the mechanisms underlying these observations. The down-regulation of gene transcripts associated with pathogen detection (tlr1, tlr8, tlr9), antigen processing (cd74a), immune cell recruitment and migration (ccl44, ccr6b, ccr7), macrophage function (csf1ra), T-cell signaling and NF-kB activation (card11, fyna, tirap) was detected in both species. These findings potentially indicate impairment in these critical early immune system processes such that both species were ultimately highly susceptible to S. iniae infection despite the detected up-regulation of transcripts typically associated with effective immune response, such as cytokines (il1β, il8, il12b2, il17rc, tnfb) and hepcidins (hamp, hamp2). The presented results collectively identify several candidate genes and associated pathways for further investigation to characterize the vulnerability of striped bass and white bass to S. iniae and that may be considered for selective breeding efforts, biotechnological intervention, and/or exploitation in the development of vaccines and alternative treatments.
Project description:Metabarcoding diets of juvenile Black Sea Bass Targeted loci environmental
| PRJNA1101022 | ENA
Project description:Discovery and validation of species-diagnostic SNP markers for black bass (Micropterus spp.): New tools for black bass conservation
Project description:Environmental variation along the geographical space can shape populations by natural selection. In the context of global warming and changing precipitation regimes, it is crucial to understand the role of environmental heterogeneity in tropical trees adaptation, given their disproportional contribution to water and carbon biogeochemical cycles. Here, we investigated how heterogeneity in freshwater availability along tropical wetlands has influenced molecular variations of the black mangrove (Avicennia germinans). A total of 57 trees were sampled at seven sites differing markedly in precipitation regime and riverine freshwater inputs. Using 2,297 genome‐wide single nucleotide polymorphic markers, we found signatures of natural selection by the association between variations in allele frequencies and environmental variables, including the precipitation of the warmest quarter and the annual precipitation. Additionally, we found candidate loci for selection based on statistical deviations from neutral expectations of interpopulation differentiation. Most candidate loci within transcribed sequences were functionally associated with central aspects of drought tolerance or plant response to drought. Moreover, our results suggest the occurrence of the rapid evolution of a population, probably in response to sudden and persistent limitations in plant access to soil water, following a road construction in 1974. Observations supporting rapid evolution included the reduction in tree size and changes in allele frequencies and in transcript expression associated with increased drought tolerance through the accumulation of osmoprotectants and antioxidants, biosynthesis of cuticles, protection against protein degradation, stomatal closure, photorespiration and photosynthesis. We describe a major role of spatial heterogeneity in freshwater availability in the specialization of this typically tropical tree.
Project description:Genome-wide DNA methylation mapping uncovers epigenetic changes associated with animal development, environmental adaptation, and species evolution. To address the lack of high-throughput methods for studying DNA methylation in non-model organisms, we developed an integrated approach for studying DNA methylation differences without a reference genome. Experimentally, our method relies on an optimized 96-well protocol for reduced representation bisulfite sequencing (RRBS), which we have validated in nine species (human, mouse, rat, cow, dog, chicken, zebrafish, carp, and sea bass). Bioinformatically, we developed the RefFreeDMA software (http://RefFreeDMA.computational-epigenetics.org) to deduce ad hoc genomes directly from RRBS reads and to pinpoint differentially methylated regions. These regions are interpreted using motif enrichment analysis and/or cross-mapping to annotated genomes. We validated our method by reference-free analysis of cell type-specific DNA methylation in the blood of human, cow, and carp. In summary, we present a cost-effective method for epigenome analysis in ecology and evolution, which enables epigenome-wide association studies in natural populations and species without a reference genome.