Wheat is one of the world's most important crops whose grain production is increasing year after year. However, its production is badly constrained by wheat rusts. Stripe rust caused by <i>Puccinia striiformis</i> f. sp<i>. tritici</i> is an important disease of wheat resulting significant yield failure in wheat growing areas around the globe. The pathogen is one of the very important yield limiting factors in Ethiopia. The severity is worse due to emergency of virulent stripe rust races at one point of the world spread to the rest of wheat producing countries by wind and human travels. Thus, development and cultivation of hereditarily diverse and tolerant varieties is the most sustainable option to overcome these diseases. The present study was carried out with the aim to identify possible sources of stripe rust resistance among Ethiopian bread wheat breeding pipelines to enhance cultivar improvement efforts and identify physiologic races involved during screening process. A total of four mono-pustule isolates were collected from Meraro and Kulumsa, stripe rust hot spot locations. Out of these, two P. striformis races; namely, PstS2 and PstS11 were identified. The former was detected at Meraro and virulent to seven of the 19-diffential lines while PstS11 displayed across Meraro and Kulumsa and virulent to nine of the19-diffential lines. Twenty-eight advanced bread wheat pipelines and a universal susceptible cultivar, Morocco were evaluated for their resistance at the seedling stage against identified stripe rust races (PstS2 and PstS11) in a controlled environment. Of the 28, twenty and seventeen lines exhibited susceptible seedling reactions to PstS2 and PstS11 with infection types ranging from 7 to 9, respectively. Those groups of lines that showed susceptible reaction at seedling stage are expected to possess poly minor genes that could be used for durable stripe rust resistance breeding in wheat. However, is advised to evaluate for adult plant resistance and postulate inherent resistance genes in these lines for fruitful recommendations.