Stripe rust is a major fungal disease of wheat. It frequently becomes epidemic in southeastern Gansu province, a stripe rust hot spot in China. Evaluations of wheat germplasm response are crucial for developing cultivars to control the disease. In total, 57 wheat cultivars and lines from Europe and other countries, comprising 36 cultivars with documented stripe rust resistance genes and 21 with unknown genes, were tested annually with multiple races of Puccinia striiformis f. sp. tritici in the field at Tianshui in Gansu province from 1993 to 2013. Seven wheat lines were highly resistant, with infection type (IT) 0 during the entire period; 16 were moderately resistant (IT 0;-2); and 26 were moderately susceptible (IT 0;-4), with low maximum disease severity compared with the susceptible control Huixianhong. 'Strampelli' and 'Libellula', with three and five quantitative trait loci, respectively, for stripe rust resistance have displayed durable resistance in this region for four decades. Ten cultivars, including 'Lantian 15', 'Lantian 26', and 'Lantian 31', with stripe rust resistance derived from European lines, were developed in our breeding program and have made a significant impact on controlling stripe rust in southeastern Gansu. Breeding resistant cultivars with multiple adult-plant resistance genes seems to be a promising strategy in wheat breeding for managing stripe rust in this region and other hot spots.