Abstract: Present in this study, random amplified polymorphic DNA(RAPD) was used to analyze polymorphisms of eight wheat yellow rust pathotypes from Gansu, and the specific RAPD markers among pathotypes were characterized. Out of 220 random 10 bp length primers were analyzed, 147 primers were giving distinct amplification bands. The genetic variations among pathotypes of wheat yellow rust were abundance. A specific 507 bp length DNA fragment was obtained from CY33 via PCR amplification with primer S301; a specific 183 bp length DNA fragment was obtained from CY32-1 with primer S39; a specific 510 bp length DNA fragment was obtained from Hybrid46-8 with primer S36; a specific 317 bp length DNA fragment was obtained from Su11-4 with primer S2140, respectively. The DNA fragment obtained from Su11-4 with primer S2140 was cloned and sequenced, specific forward and revise primers were designed for Su11-4 and the sequence-characterized amplified region SCAR marker was developed as well. The results indicate that a molecular detection system for Puccinia striiformisf.sp tritici could be established by mean of searching for specific RAPD fragments among pathotypes. The results also suggest that development of SCAR marker would be a important approach to accurately detect pathotype Su11-4 of Puccinia striiformisf. sp tritici.