The genetic diversity and differentiation of 147 Aegilops tauschii accessions from Henan, Shannxi, Xinjiang in China and the Middle East, were investigated by SSR assay with 32 pairs of primers homogeneously distributed on 14 chromosomes of Ae. tauschii. The results demonstrated that 26 loci were polymorphic, with an average of 4.15 alleles on each locus. Nei's genetic diversity index (He) and polymorphism information content (PIC) value were 0.243 and 0.226, respectively. Genetic variations among different populations varied greatly. The maximal diversity (values were found in Middle East population (He =0.607, PIC =0.551), which was much higher than those in Shannxi (He =0.055, PIC =0.047) and Henan (He =0.024, PIC =0.021) populations. Analysis of molecular variance (AMOVA) showed that the genetic differentiation among the geographical populations was significant (P<0.05) and accounted for about 52% of the total genetic variation. Meanwhile，the genetic differentiation was obvious between the populations of Henan and Shannxi(Fst =0.210). These results would offer useful information and evidence for further understanding the origin and evolution of the Ae. tauschii from China.