Abstract: In order to understand the mitochondrial genome structure of Luciola kagiana Matsumura, 1928（Coleoptera: Lampyridae） and phylogenetic analysis of Luciolinae, the mitochondrial genome of L. kagiana was sequenced, and the genome sequences were then assembled, annotated and characterized. The results showed that the complete mitochondrial genome of L. kagiana was 16 317 bp in length, including 13 proteincoding genes（PCGs）, 22 tRNA genes, 2 rRNA genes, and one control region. At the same time, the composition and structure of the mitochondrial genomes of 21 fireflies（including 19 species belonging to the subfamily Luciolinae and 2 species belonging to the subfamily Lampyrinae） were analyzed. It was found that the basic features of mitochondrial genome composition and structure remained consistent in Lampyridae, but there was some variability under subfamilies in whole genome, length of control region, A+T skew and intergenic sequences. And the phylogenetic relationship was constructed by using maximum likelihood method based on the 13 PCGs of 21 species of fireflies. The results showed that L. kagiana gathered together with other two firefly species of Luciola on a branch, and the fireflies of Luciolinae and Lampyrinae each gathered together on a branch, and the fireflies of the same genus under the Luciolinae were clustered on a branch, respectively. This study shows that the development of molecular classification technology can assist and promote the systematization and accuracy of traditional morphological classification.