The stomatal complex on the surface of maize leaves is closely related to photosynthesis and transpiration, and the study of maize stomatal phenotypes and the discovery of their regulatory genes are of great importance for the breeding of high-quality and high-yielding maize. In this study, rapid scanning electron microscopy was used to obtain images of the abaxial stomata of 457 maize inbred lines with extensive genetic variation, and stomata density was obtained by counting. The results of correlation showed that stomata density was significantly correlated with leaf width, and Analysis of variance found that there were significant differences (P value < 0.05) in stomata density among different leaf width and 100-grain weight. The highest stomata density was found in the inbred lines with wide and short leaves and higher 100-grain weight. Furthermore, genome-wide association analysis was performed using a mixed linear model. It showed that eight SNPs significantly associated with stomata density were obtained, which could explain 35.507% of the phenotypic variation. Among these, four SNPs on chromosome 5 were tightly linked, mainly formatting two haplotypes, CTTA (0.636) and TCCG (0.330). Twelve genes with functional annotation were identified within 100 kb upstream and downstream of the eight SNPs. One gene, GRMZM2G068277, which had been shown to be involved in plant mitotic processes and exhibited high expression at the leaf base, was therefore the most likely candidate gene for stomata density. The results presented here could provide references for further cloning of functional genes related to stomata density.