Abstract: 339 hexaploid Triticale from International Maize an d Wheat Improvement Center （CIMMYT）were analyzed using 100 microsatellite markers distributed across the whole Triticale genome. 100 pairs of primers detected 323 alleles, ranged from 1 to 6, with an average abundance of allelic variation was 3.23; polymorphic information content (PIC) ranged from 0 to 0.748, an average of PIC was 0.465, belong to moderate polymorphic loci; average genetic diversity index (H') was 0.1439, indicating that the genetic diversity of hexaploid Triticale was relatively rich. Clustering results based on Nei's genetic distance showed that the tested materials can be divided into six groups. Among them, genetic distance between groups Ⅵ and others was very obvious.