Abstract:The complete mitochondrial genome of Acrossocheilus wenchowensis was obtained by PCR based on 8 primers, which were designed on the basis of published fish mitochondrial genome (GenBank accession number: KC_495074). The complete mitochondrial genome was 16 591 bp in length, including 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and a non-coding control region. The organization and location of genes in the mitochondrial genome of A.wenchowensis were consistent with Cyprinidae fishes published in GenBank. Of 37 genes, 1 protein-coding gene(ND6) and 8 tRNA genes(tRNAGln, tRNAAla, tRNAAsn, tRNACys, tRNATyr, tRNASer(UCN), tRNAGlu and tRNAPro) were encoded on the L strand, the remainders on the H strand. The nucleotide composition was 30.93% (A), 24.86% (T), 16.41% (G) and 27.81% (C). Most tRNA genes could form typical secondary structures except tRNASer(AGN). The COⅠ gene used GTG as an initiation codon, with the remaining 12 using ATG of total 13 protein genes, 10 genes had complete stop codons,and COⅡ, ND4 and Cytb had an incomplete stop codon T. The phylogenetic analysis of A.wenchowensis on Cyprinidae fishes was made from the level of mitochondrial genome based on amino acids sequence of all protein-coding genes in 18 species of 17 genera, and the complete mitochondrial genome of A.wenchowensis will help to provide basic data for the classification of Acrossocheilu and Cyprinidae fishes.