Wheat (Triticum aestivum) rust outbreak especially yellow rust is the primary production constraint in Ethiopia. Arsi Zone are hot spots areas for yellow rust (Puccinia striiformis (Pst)) particularly Meraro and Bekoji sites are frequently epidemic of yellow rust was occur due to higher pressure of inoculums in the areas. The objective of this study was to identify sources of resistance against yellow rust at adult plant stage under hot spot sites. Three hundred seventy five elite breeding lines, selected and advanced at Meraro, Arsi-Robe and Bekoji were planted in yellow rust hot spot areas of the country under natural infections. Yellow rust scores of 34,10 and 54% of the line at Meraro, 61, 23.5 and 15.5% of lines at Arsi-Robe and 35.5, 21.3 and 43.2% lines at Bekoji were resistant to moderately resistant, intermediate and moderately susceptible to susceptible category, respectively. Overall coefficient of infection (ACI) at Meraro and Bekoji was greater than Arsi-Robe that is indicating higher disease pressure in Meraro and Bekoji than that of Arsi-Robe. Among the lines, 34%, 35.5% and 61% at Meraro, Bekoji and Arsi-Robe showed good levels of resistance, with a severity less than 20% respectively. However, only 43.7% of the lines showed a good level of resistance (<30% severity) in all locations. These lines could have resistance to the current yellow rust diseases pressure at all testing sites Generally, most of the lines, which showed better resistance at Arsi-Robe were susceptible at Meraro and Bekoji.