Leveraging Short-Read Sequencing to Explore the Genomics of Sepiolid Squid

Synopsis Due to their large size (∼3–5 Gb) and high repetitive content, the study of cephalopod genomes has historically been problematic. However, with the recent sequencing of several cephalopod genomes, including the Hawaiian bobtail squid (Euprymna scolopes), whole-genome studies of these molluscs are now possible. Of particular interest are the sepiolid or bobtail squids, many of which develop photophores in which bioluminescent bacterial symbionts reside. The variable presence of the symbiosis throughout the family allows us to determine regions of the genome that are under selection in symbiotic lineages, potentially providing a mechanism for identifying genes instrumental in the evolution of these mutualistic associations. To this end, we have used high-throughput sequencing to generate sequence from five bobtail squid genomes, four of which maintain symbioses with luminescent bacteria (E. hyllebergi, E. albatrossae, E. scolopes, and Rondeletiola minor), and one of which does not (Sepietta neglecta). When we performed K-mer based heterozygosity and genome size estimations, we found that the Euprymna genus has a higher predicted genome size than other bobtail squid (∼5 Gb as compared to ∼4 Gb) and lower genomic heterozygosity. When we analyzed the repetitive content of the genomes, we found that genomes in the genus Euprymna appear to have recently acquired a significant quantity of LINE elements that are not found in its sister genus Rondeletiola or the closely related Sepietta. Using Abyss-2.0 and then Chromosomer with the published E. scolopes genome as a reference, we generated E. hyllebergi and E. albatrossae genomes of 1.54–1.57 Gb in size, but containing over 78–81% of eukaryotic single-copy othologs. The data that we have generated will enable future whole-genome comparisons between these species to determine gene and regulatory content that differs between symbiotic and non-symbiotic lineages, as well as genes associated with symbiosis that are under selection.