Karnal bunt (KB) of wheat, caused by Tilletia indica (Mitra) Mundkur, adversely affects international wheat trading and the movement of germplasm between countries due to quarantine restrictions. Breeding for host plant resistance requires the identification of KB resistance sources. Accessions of the D genome progenitor of bread wheat, Aegilops tauschii, were screened in a specially designed screen-house, where the optimum environmental conditions conducive for KB development were simulated by controlling temperature, humidity, fogging and shading. The 183 accessions were subjected to artificial inoculation with a mixture of nine KB isolates, and seven proved highly resistant and four moderately resistant over three rounds of screening over 3 years.