Sesame charcoal rot is an important disease affecting sesame production in China, seriously threatening both yield and quality. To explore resistant germplasm resources, a total of 420 sesame accessions (including 27 introduced from abroad, 212 landraces and 181 modern varieties) were evaluated for resistance under field conditions in Pingyu County, Henan Province, over two consecutive years (2023–2024). The disease index (DI) and resistance index (RI) were used for systematic evaluation. The results showed that no immune (I) accessions were identified. Twenty accessions (4.77%) were consistently classified as highly resistant (HR) over the two years, while 17 accessions (4.06%) were consistently highly susceptible (HS). A total of 112 accessions (26.73%) showed stable moderate resistance (MR) or above and 182 accessions (43.44%) showed stable moderate susceptibility (MS) or below. The average DI of modern varieties was significantly lower than that of introduced germplasm and landraces, and the proportions of HR and MR materials were higher, indicating that artificial selection has effectively improved resistance levels. Among different ecological regions, the Huang-Huai summer sesame region had the highest proportion of HR germplasm (34.5% in 2023, 12.5% in 2024), while the Northeast and North China spring sesame regions had higher proportions of HS materials. Cluster analysis divided the 420 accessions into four groups: HR, MR, MS and HS. Combining both evaluation methods, a set of excellent resistant germplasm accessions were identified, such as Zhu 044, Xixiangxian Zhima, Wanzhi 24 and Fuzhi 7, providing important germplasm resources and a theoretical basis for genetic improvement of charcoal rot resistance and breeding of resistant sesame varieties.