High-quality reference genome of the African hermit spider, Nephilingis cruentata, and sex chromosome evolution in spiders

Background: Chromosome-level genome assemblies are increasingly enabling tests of chromosome evolution, conserved synteny, and sex chromosome conservation across diverse animal lineages, including spiders. Results: Here, we present a chromosome-level genome assembly for the African hermit spider, Nephilingis cruentata, a species with extreme female-biased sexual size dimorphism and a cytogenetically inferred XX2 sex chromosome system. The final Hi-C-assisted assembly spans 1.72 Gbp, with 99.5% of bases assigned to 13 pseudochromosomes, a scaffold N50 of 131.6 Mbp, and a BUSCO completeness score of 98.8%. We annotated 20,021 protein-coding genes, and repetitive elements accounted for 42.7% of the genome. Sex-specific whole-genome resequencing identified Chr02 and Chr07 as candidate X chromosomes based on reduced male coverage, consistent with the expected XX2 system. Using comparative whole-genome alignments across existing chromosome-scale spider assemblies, we also show that sex-linked chromosomes retain broad homologous identity across sampled spider lineages but exhibit lower synteny conservation and greater chromosome-length divergence than autosomes. Conclusions: These results suggest that spider sex chromosomes are conserved in homologous identity but more labile in structure, providing a comparative framework for studying sex chromosome conservation and divergence across Araneae.