Abstract: The high-throughput sequencing of illumina platform was used to sequence the whole mitochondrial genome of Rana kukunoris. Then, the mitochondrial genome was assembled and annotated. The total length of the sequence was 21913 bp, which mainly included four parts: tRNA gene (22), rRNA gene (2), CDS (13) and D-loop (1). ExceptNAD6in the light chain, other protein coding genes were on the same chain. Of the coding genes, three overlapping genes were found:COX1/trnS2,ATP6/ATP8andNAD4L/NAD4. Mitochondrial genome of Rana kukunoris had AT preference. Leu and Ser were the most frequently used amino acids in protein coding genes, and all tRNA had the classic clover structure. KaKs analysis showed that the coding genes of mitochondrial genome were not significantly positively selected in Rana kukunoris. The phylogenetic tree constructed based on the whole mitochondrial genome showed that Rana kukunoris was closely related to Rana chensinensis. It provided the genetic information of Rana kukunoris mitochondrial genome and aid future investigation of phylogenetics, speciation and evolution of Rana genus.

Key words: Rana kukunoris, mitochondrial genome, sequence analysis, phylogeny, species differentiation