Abstract: Harmonia axyridis, one of the most important insect predators, has been widely used in biological control in agriculture. In this study, the mitochondrial COII gene was used as genetic marker to investigate the genetic structure and diversity of H. axyridis geographic populations distributed in Shaanxi Province. Level of genetic differentiation and gene flow among different populations of H. axyridis were analyzed. For all the 529 H. axyridis individuals from 21 populations, 15 haplotypes were observed (named as Hap1-Hap15). Hap1 and Hap2 were dominant haplotypes in all sampled individuals, accounting for 34.4% and 37.6% respectively. The overall haplotype diver-sity was 0.732, and it ranged from 0.652 to 0.786 within population. The average gene flow (N_m) among 21 populations was 10.13, and the overall F_ST value was 0.024, implying low genetic diffe-rentiation among all populations. Results from neutrality tests suggested that H. axyridis populations followed the neutral evolution model and the population size remained stable. Molecular variance analysis (AMOVA) showed high proportion of total genetic variance attributed to variations within populations. Phylogenetic tree based on Nei’s genetic distance of different geographic populations showed clear divergence between northern, central, and southern Shaanxi groups. Results from Mantel test suggested that the genetic distance was weakly correlated with geographical distance between populations. Geographical pattern of genetic structure and diversity in populations from different regions indicated weak gene flow between northern, central, and southern Shaanxi due to Qinling Mountains barrier and climate difference.