[Objectives]  To clone the cDNA sequences of the A, B, C and D subunit genes of the Spodoptera frugiperda V-ATPase V₁ region and compare the expression of these genes in different developmental stages in order to both improve the prevention and control of S. frugiperda and identify new target genes for pesticides. [Methods]  The sequences of the genes encoding the V-ATPase subunits A, B, C and D were cloned with PCR (polymerase chain reaction), and the expression of each gene was analyzed in different development stages with real-time quantitative PCR. [Results]  The A, B, C and D subunit genes of the V₁ region are highly conserved in the Lepidoptera. All four genes were expressed during the entire developmental process but were more highly expressed in the larval and adult stages, suggesting that V-ATPase plays an important role in the growth and development of larvae and in adult reproduction. The expression of the four subunit genes differed in different development stages, suggesting that each has different functions. [Conclusion]  The four V-ATPase V₁ subunit A, B, C and D genes had different expression levels in different developmental stages of S. frugiperda, which suggests that they play an important role in regulating the growth, development and reproduction of this species.