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Gordon, L.

Publications and source records attributed to Gordon, L..

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Using long-read sequencing to detect imprinted DNA methylation

Systematic variation in the methylation of cytosines at CpG sites plays a critical role in early development of humans and other mammals. Of particular interest are regions of differential methylation between parental alleles, as these often dictate monoallelic gene expression, resulting in parent of origin specific control of the embryonic transcriptome and subsequent development, in a phenomenon known as genomic imprinting. Using long-read nanopore sequencing we show that, with an average genomic coverage of approximately ten, it is possible to determine both the level of methylation of CpG sites and the haplotype from which each read arises. The long-read property is exploited to characterise, using novel methods, both methylation and haplotype for reads that have reduced basecalling precision compared to Sanger sequencing. We validate the analysis both through comparison of nanopore-derived methylation patterns with those from Reduced Representation Bisulfite Sequencing data and through comparison with previously reported data. Our analysis successfully identifies known imprinting control regions as well as some novel differentially methylated regions which, due to their proximity to hitherto unknown monoallelically expressed genes, may represent new imprinting control regions.

genomics

Mapping Social Ecological Systems Archetypes

While sustainable development goals are by their nature global, their achievement requires local action and thus targeting and monitoring sustainable solutions tailored to different social and ecological contexts. Ostrom stressed that there are no panaceas or universal solutions to environmental problems, and developed a social-ecological systems (SES) framework -a nested multi-tier set of variables- to help diagnose problems, identify complex interactions, and solutions tailored to each SES arena. The framework has been applied to over a hundred cases, typically reflecting in-depth analysis of local case studies, but with relatively small coverage in space and time. While case studies are context rich and necessary, it can be difficult to upscale their lessons to policy making realms. Here we develop a data driven method for upscaling Ostroms SES framework and apply it to a context where data is scarce, incomplete, but also where sustainable solutions are needed. The purpose of upscaling the framework is to create a tool that facilitates decision-making on data scarce contexts such as developing countries. We mapped SES by applying the SES framework to poverty alleviation and food security issues in the Volta River basin in Ghana and Burkina Faso. We found archetypical configurations of SES in space. Given data availability, we study their change over time, and discuss where agricultural innovations such as water reservoirs might have a stronger impact at increasing food security and therefore alleviating poverty and hunger. We conclude by outlining how the method can be used in other SES comparative studies.

ecology

De novo assembly, characterization, functional annotation and expression patterns of the black tiger shrimp (Penaeus monodon) transcriptome

The black tiger shrimp (Penaeus monodon) remains the second most widely cultured shrimp species globally. However, issues with disease and domestication have seen production levels stagnate over the past two decades. To help identify innovative solutions needed to resolve bottlenecks hampering the culture of this species, it is important to generate genetic and genomic resources. Towards this aim, we have produced the most complete publicly available P. monodon transcriptome database to date. The assembly was carried out in multiple assemblers using 2x125 bp HiSeq data from PolyA selected, ribo-depleted RNA extracted from nine adult tissues and eight early life-history stages. In total, approximately 700 million high-quality sequence reads were obtained and assembled into 236,388 clusters. These were then further segregated into 99,203 adult tissue specific clusters, and 58,678 early life-history stage specific clusters. The final transcriptome had a high TransRate score of 0.37, with 88% of all reads successfully mapping back to the transcriptome. BUSCO statistics showed the assembly to be highly complete with low fragmentation, few genes missing, but higher redundancy or transcript duplication (Complete: 98.2% (Duplicated: 51.3%), Fragmented: 0.8%, Missing: 1.0%), and to greatly exceed the completeness of existing P. monodon transcriptomes. While annotation rates were low (approximately 30%), as is typical for a non-model organisms, annotated transcript clusters were successfully mapped to several hundred functional KEGG pathways. To help address the lack of annotation, transcripts were clustered into groups within tissues and early life-history stages, providing initial evidence for their roles in specific tissue functions, or developmental transitions. Additionally, transcripts of shrimp viruses previously not known to occur in Australia were also discovered. We expect the transcriptome to provide an essential resource to investigate the molecular basis of commercially relevant-significant traits in P. monodon and other shrimp species.

molecular biology