An isolate of Puccinia coronata f.sp. avenae with virulence against the oat crown rust resistance gene Pc94 was discovered in oat fields in Córdoba, Spain. In order to identify alternative sources of resistance to this virulent isolate, a collection of 159 Avena accessions, including 107 A. sativa and 29 A. byzantina landraces and 23 commercial A. sativa and A. strigosa cultivars, was screened. Eight resistant landraces and four cultivars were selected according to macroscopic assessment to further characterize the operative defence mechanisms. Histological studies showed a range of defence mechanisms, acting alone or in combination, which impeded fungal development at different stages. Some accessions allowed reduced fungal growth before mesophyll penetration. In others, the fungus was arrested at the penetration stage by mesophyll cell wall strengthening and/or papilla deposition. Mesophyll cells of several accessions were penetrated by the fungus, but then the hypersensitive response (HR) leading to cell death hampered fungal development. In some cases cell death was very fast and colonies aborted early, whereas in other cases necrosis was observed later and associated with numerous secondary hyphae, suggesting a slow HR. Characterization of defence mechanisms will be useful for breeding programmes and for further cellular and molecular studies to unravel the bases of resistance. Commonalities with the resistance of the same oat collection to powdery mildew are discussed.