Abstract: This study aimed to elucidate the intra- and inter-population genetic variation of Pastor roseus in Xinjiang, China. Sequences of the mitochondrial genes COI and Cytb of 108 individuals from 10 distinct geographical populations across four regions of Xinjiang Uyghur Autonomous Region were analyzed. The mitochondrial genes were 1551 bp and 1143 bp in full length, respectively, and the AT content of bases was greater than the GC content. Based on the molecular variation in COI and Cytb, 62 and 69 haplotypes were detected, respectively; the average haplotype diversity (Hd) values were 0.976±0.006 and 0.944±0.018, respectively, and the nucleotide diversity (π) values were 0.00316±0.00016 and 0.00292±0.00021, respectively, indicating that there was high genetic diversity among the 10 population. Analysis of molecular variance (AMOVA) indicated that the major source of genetic variation was within the populations. Analysis of molecular signatures of neutrality indicated that Tajima’s D value was not significant, but Fu’s FS was significant, suggesting that P. roseus populations have recently experienced a large population expansion, but that the populations are currently relatively stable and the selection pressure is low. The results of the study not only provide a theoretical basis for the conservation of P. roseus, but also help to utilize this species in the comprehensive management of grassland locusts.