Leaf stripe disease, a seed-borne fungal disease caused by Pyrenophora graminea, poses a significant threat to hulless barley (Hordeum vulgare var. nudum) production on the Qinghai-Tibet Plateau. This study aimed to identify genetic factors conferring resistance to leaf stripe by analyzing an F3 population derived from a cross between the resistant landrace Teliteqingke and the susceptible landrace Dulihuang. Genetic analysis revealed that resistance in Teliteqingke was governed by two dominant genes. Using bulked segregant analysis combined with SNP array (BSA-SNP) and RNA-seq, we identified two candidate regions on chromosomes 3H and 7H. Further analysis focused on chromosome 3H, which revealed a candidate genomic region containing seven potential disease-resistance genes. Among these, RT-qPCR experiments demonstrated that HORVU.MOREX.r3.3HG0232110.1 (encoding a RING/U-box superfamily protein) and HORVU.MOREX.r3.3HG0232410.1 (encoding a bZIP transcription factor) showed significant expression induction following inoculation with P. graminea. These genes can be candidate genes involved in resistance mechanism against leaf stripe in Teliteqingke. These results provide a foundation for functional validation of these genes and offer valuable insights for breeding disease-resistant hulless barley.