Gray leaf spot caused by Stemphylium spp., is a major disease of tomatoes, and it threatens its cultivation worldwide, especially in warm and humid areas. This study was conducted on 223 tomato germplasm conserved at the National Agrobiodiversity Center to select the resources resistant to the gray leaf spot pathogen strain previously isolated in Korea, using a bioassay and genotypic analysis of the resistance gene (Sm). Two weeks after inoculation with Stemphylium lycopersici, the disease index (rated on a scale of 0-4) of gray leaf spot was assessed in detached tomato leaves. The results showed that 22 resources were resistant, with a disease index of 0-1. Additionally, 65 genetic resources were found to be moderately resistant, with a disease index between 1.0 and 2.0. Subsequently, Hybridization Probe Melting (HPM) analysis of the 22 resistant genetic resources confirmed the genotype of the gray leaf spot resistance gene (Sm). Among them, 20 genetic resources showed a homozygous resistant genotype. The resources selected in this research may contribute to the breeding of new tomato varieties resistant to gray leaf spot and may serve as a basis for further genotypic analysis studies.