【Aim】 This study aims to identify Parastagonospora pseudonodorum in barley imported from French and evaluate its pathogenicity. 【Method】 A fungal isolate 5225-17 was obtained from the barley sample imported from French by the plate culture method, and it was identified based on colony morphology as well as ITS, β-tubulin, and β-xylosidase sequences. Furthermore, the pathogenicity of this isolate was tested. 【Result】 The colony of 5225-17 cultured on potato dextrose agar (PDA) appeared white flocculent in the front view and light pink in the center in the back view, with a regular margin. The isolate experienced rapid growth at a rate of 0.5~1.0 cm·d^-1. It showed low sporulation on PDA and sporulation on water agar after 21 days of incubation at 20 ℃. The conidia were cylindrical, transparent, slightly curved or straight at the upper end, blunt or round at the end, with 3~5 septa, oil balls, and the average size of 26.8 μm×3.6 μm. The asci were rod-shaped, transparent, with a round top, (40~80) μm×(8~11) μm. Each ascus contained 8 ascospores, being light yellow to light yellowish brown, spindle-shaped, straight or slightly curved, obviously constricted at septa, with 3 septa, occasionally 2 or 4, and average size of 20.8 μm×4.1 μm. The ITS, β-tubulin, and β-xylosidasesequences of the isolate were identical to those of Parastagonospora pseudonodorum. The phylogenetic tree built based on multiple genes indicated that isolate 5225-17 shared the same clade with P. pseudonodorum and had a distant genetic relationship with P. nodorum. The pathogenicity test showed that isolate 5225-17 caused typical symptoms on both detached and living leaves. 【Conclusion】 The isolate 5225-17 is identified as P. pseudonodorum.