The first mitogenomes of the subfamily Epipleminae (Lepidoptera, Uraniidae) and phylogenetic analysis of Macroheterocera

The subfamily Epipleminae, the largest group within Uraniidae (Lepidoptera, Macroheterocera, Geometroidea), comprises small, nocturnal moths primarily distributed in tropical regions. Taxonomic and molecular phylogenetic studies of Epipleminae have long been challenging, and this group remains genetically understudied, with no mitochondrial genomes (mitogenomes) reported to date, despite their wide utility in phylogenetic research. Here, we sequenced, assembled, and annotated the first complete mitogenomes of four epiplemine species: <i>Dysaethria flavistriga</i> (15,404 bp), <i>Monobolodes prunaria</i> (15,258 bp), <i>Phazaca alikangensis</i> (15,482 bp), and <i>Warreniplema fumicosta</i> (15,467 bp), using high-throughput sequencing technology. These mitogenomes exhibit typical gene arrangement of ditrysian Lepidoptera, along with distinctive features such as rare (TA)n microsatellite repeats in the 16S rRNA. Most protein-coding genes (PCGs) initiate with standard ATN start codons and terminate with TAA or a single T residue. Codon usage analysis revealed UUA (Leu2), AUU (Ile), UUU (Phe), AUA (Met), and AAU (Asn) as the five most frequently used codons. All tRNAs display canonical cloverleaf secondary structures, except for trnS1, which lacks the DHU arm. Comprehensive phylogenetic analyses that incorporated existing macroheteroceran mitogenomic data provided robust support for the placement of Epipleminae under Uraniidae, offered the first mitogenome-based evidence supporting the monophyly of Geometroidea on family level, and strongly supported a sister relationship between Geometroidea and Noctuoidea.