Abstract
(1) Background: The insect mitogenome encodes essential genetic components and serves as an effective marker for molecular identification and phylogenetic analysis in insects due to its small size, maternal inheritance, and rapid evolution. The morphological identification of Ablabesmyia is challenging, particularly for non-experts. Thus, there is an increasing need for molecular data to improve classification accuracy and phylogenetic analysis. (2) Methods: Our analysis encompassed eight species of Ablabesmyia, a single species of Conchapelopia, one species of Denopelopia, and one species of Thienemannimyia, all originating from China. We then performed a comprehensive analysis of the nucleotide composition, sequence length, and evolutionary rate. (3) Results: All newly assembled mitogenomes displayed a negative GC-skew, indicating a cytosine bias, while most exhibited a positive AT-skew, reflecting an adenine and thymine abundance. All thirteen protein-coding genes (PCGs) featured the conventional start codon ATN, aligning closely with the typical mitochondrial start codon observed in insects. The evolutionary rates of these PCGs can be ordered as follows: ND2 > ATP8 > ND6 > ND4 > ND5 > ND3 > ND4L > ND1 > CYTB > COIII > ATP6 > COII > COI. (4) Conclusions: These newly sequenced mitogenomes exhibit structural features and nucleotide compositions that closely align with those of previously reported Chironomidae species, marking a significant expansion of the chironomid mitogenome database.