Characterization of the Complete Mitochondrial Genome and Evaluation of COI Barcoding in Philonis inermis (Coleoptera: Curculionidae: Cryptorhynchinae) Using Genome Skimming

<ns7:p> Background <ns7:italic>Philonis inermis</ns7:italic> is a Neotropical stem-galling weevil specialized on the invasive vine <ns7:italic>Passiflora foetida</ns7:italic> and represents a promising candidate for biological control. However, no genomic or barcoding data have previously been available for this genus, limiting its taxonomic resolution and risk assessment potential. Methods We used shallow whole-genome sequencing of two individuals reared under controlled conditions to assemble the complete mitochondrial genome of <ns7:italic>P. inermis.</ns7:italic> The mitogenome was annotated and compared with other Cryptorhynchinae species. Cytochrome c oxidase subunit I (COI) sequences from 20 Colombian individuals were analyzed alongside 24 American Cryptorhynchinae barcodes retrieved from GenBank to evaluate intra- and interspecific genetic divergence. Results The <ns7:italic>P. inermis</ns7:italic> mitogenome is 15,120 bp in length, AT-rich (77.0%), and contains 36 genes, including 13 protein-coding genes, 21 tRNAs, and two rRNAs. The tRNA-Ile was not detected, likely obscured within the variable control region, as reported for other cryptorhynchines. Phylogenetic analysis based on mitogenomic sequences placed <ns7:italic>P. inermis</ns7:italic> as a well-supported clade closely related to <ns7:italic>Eucryptorrhynchus.</ns7:italic> COI barcode analysis revealed extremely low intraspecific divergence (pairwise K2P ≤ 0.006) and a pronounced barcode gap distinguishing <ns7:italic>P. inermis</ns7:italic> from other Cryptorhynchinae species. In addition, 196 single-copy orthologs were identified from the shallow assemblies, providing a basis for future phylogenomic investigations. Conclusion This study presents the first complete mitochondrial genome for the genus <ns7:italic>Philonis</ns7:italic> and confirms the reliability of COI barcoding for its accurate identification. These genomic resources lay the foundation for integrative taxonomic, comparative, and evolutionary studies, and support the evaluation of <ns7:italic>P. inermis</ns7:italic> as a potential biological control agent against <ns7:italic>P. foetida.</ns7:italic> </ns7:p>