A new piece in the repeatome puzzle of Triatominae bugs: The analysis of Triatoma rubrofasciata reveals the role of satellite DNAs in the karyotypic evolution of distinct lineages.

The genome of Triatoma rubrofasciata, a representative of the North American Triatomini lineage, was analysed to characterise its repetitive DNA content and satellite DNA (satDNA) organisation. Using RepeatExplorer2, we determined that repetitive elements comprise approximately 25% of the genome in a male sample from Vietnam and 16% in a female sample from China, with satDNA being the most abundant component. The satellitome analysis revealed 126 satDNA families in the male and 114 in the female, with marked quantitative differences driven by the amplification of two satDNA families: TrubSat001-166 and TrubSat002-9. Fluorescence in situ hybridization (FISH) confirmed that TrubSat002-9 is enriched in the Y chromosome, explaining its lesser abundance in the female genome. Chromosomal mapping revealed three distribution patterns of satDNA: (i) localisation in autosomal heterochromatin, (ii) restriction to the Y chromosome, and (iii) presence in euchromatin. SatDNA landscapes showed sharp peaks at low divergence values, consistent with recent amplifications in heterochromatic regions, and broader peaks at higher divergence levels, suggesting older satDNAs located in euchromatic regions. Additionally, several satDNA families are conserved among T. rubrofasciata, T. infestans, T. delpontei and Rhodnius prolixus, supporting the "library hypothesis" of satDNA evolution. Our findings highlight the differential amplification of satDNA families linked to heterochromatin expansion, particularly in autosomes, and the conservation of Y-linked repeats. This study provides new insights into the dynamic role of satDNAs in the karyotypic evolution of Triatominae bugs.