Abstract: 【Objective】 This study aimed to analyze and investigate the genetic diversity of Vespa soror based on the mitochondrial cytochrome b （Cytb） gene， clarify the current status of genetic diversity and differentiation of Vespa soror populations in China， thereby providing a theoretical basis for conserving and sustainably utilizing Vespa soror resources.【Method】 Vespa soror samples were collected from 15 geographic populations across seven provinces or autonomous regions in China， followed by total genomic DNA extraction and PCR amplification. The gene sequence characteristics of the mitochondrial Cytb gene of Vespa soror were analyzed via sequencing， on the basis of which analyses of haplotype and genetic diversity， genetic differentiation and gene flow， mismatch distribution and molecular variance were performed to establish haplotype network diagrams and phylogenetic trees.【Result】 The length of clean reads of Cytb gene was 393 bp， with 15 variable sites， and the AT content of Cytb gene （69.9%） was higher than GC content （30.1%）， showing a marked preference for A/T bases. A total of 13 haplotypes were identified from the 15 Vespa soror populations， and H1 was the dominant haplotype， distributed in 8 populations， showing the highest occurrence frequency of 42.72%. For the five populations， the overall haplotype diversity index was 0.765， the nucleotide diversity index was 0.00614， and the average number of nucleotide differences was 2.415. Among the 105 comparison groups of the 15 populations， the fixed coefficient （F_ST） index of the 74 comparison groups was above 0.25， taking up 70.48% of the total comparison group； the gene flow （N_m） of the 80 comparison groups<1.00， accounting for 76.19%. The observed values of the mismatch distribution curve of the populations showed a distinct multimodal distribution. Genetic variation mainly existed among populations， with a contribution rate of 68.75586%.【Conclusion】 Vespa soror populations showed high genetic diversity， with high haplotype and nucleotide diversity indexes. Genetic differentiation between populations is obvious， and most pairwise populations show high levels of genetic differentiation and generally low levels of gene flow. Genetic variation is mainly attributed to differences among populations. The haplotype network has a radial structure， and the phylogenetic tree forms four major clades， showing a distinct pattern of geographic cluster and differentiation.