Abstract
The dung beetle family Geotrupidae (Scarabaeoidea) plays a vital ecological role in nutrient cycling and soil health, yet the scarcity of complete mitochondrial genome (mitogenome) data has hindered phylogenetic and comparative studies within this family. Here, we sequenced, assembled, and annotated the first complete mitogenomes of Geotrupes stercorarius and Phelotrupes auratus, collected from the Qinghai-Tibetan Plateau. Comparative analysis of these two novel mitogenomes with eight existing mitogenomes revealed conserved architectural features across Geotrupidae, such as gene arrangement, tRNA secondary structures, and small intergenic spacers. Nucleotide composition was largely conserved, though marked divergence occurred at the third codon positions. Substantial structural variation was observed in non-coding regions, particularly in the control region and the nad2-trnW spacer. Evolutionary analyses indicated strong purifying selection across all protein-coding genes, with no evidence of widespread positive selection linked to high-altitude adaptation. Phylogenetic reconstruction consistently recovered the relationships (Bolboceratinae, (Lethrinae, Geotrupinae)), with Anoplotrupes and Geotrupes forming sister genera within Geotrupinae. This study provides additional mitogenomic resources and a well-supported phylogenetic framework for Geotrupidae, resolving key taxonomic uncertainties and establishing a basis for future evolutionary and ecological research.