Molecular phylogeny of selected North American Chrysobothris species (Coleoptera: Buprestidae) inferred from assembled mitochondrial genomes

The genus Chrysobothris (Coleoptera: Buprestidae) has approximately 134 species occurring in North America north of Mexico. Several are considered pests of economically important forest, nut, fruit, and landscape trees. Members of the Chrysobothris femorata species complex are morphologically similar, posing significant challenges for accurate identification. This challenge highlights the need for rapid and reliable species identification to develop species-specific IPM strategies. We applied next-generation methods to differentiate 12 taxa within the C. femorata group and to evaluate taxonomic relationships across Chrysobothris and closely related genera. Adult beetle specimens from field collections, private collections, and museums were analyzed. We optimized non-destructive DNA extraction methods to preserve rare museum specimens. Illumina sequencing data from selected specimens were used to assemble mitochondrial genomes with GetOrganelle. Genome annotations were performed using MITOS2, and circular genome maps were generated with OGDRAW. While NCBI currently lists nucleotide information for 26 Chrysobothris species based mostly on partial COI sequences, we provide complete mitochondrial genomes for 56 Chrysobothris species and five closely related buprestid taxa. These mitogenomes average ~15,000 base pairs in length and include sequence data for 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), and 2 ribosomal RNAs (rRNAs). A maximum likelihood phylogenetic reconstruction was performed using IQ-TREE2, incorporating all 37 mitochondrial genes from each sampled Chrysobothris species and representative outgroups. The resulting phylogeny is expected to clarify species relationships within the C. femorata group and support accurate identification, ultimately aiding in the development of improved pest management strategies for nursery production systems.