[Objective] The aim was to study on the genetic diversity of local varieties of Chinese Hu mulberry (Morus L.). [Method] The genetic diversity of 141 copies of Hu mulberry varieties was analyzed by ISSR molecular markers. [Result] 12 ISSR primers had amplified a total of 90 amplified, of which 57 bands were polymorphic, and the polymorphic rate was 63.33%. The genetic similarity coefficients of 141 Hu mulberry germplasm resources varied from 0.633 3 to 1.000 0 with the average of 0.483 35, indicating that there was difference on genetic diversity among different varieties of Hu mulberries. A dendrogram of all 141 Hu mulberry varieties based on the genetic similarity coefficients using ISSR molecular markers was generated by UPGMA cluster method. Clustering of the 141 Hu mulberry varieties did not correspond with the conventional classification involving differences in style, leaf, branch, fruit and other morphological or agronomical characters. [Conclusion] Four subgroups clearly represented the genetic relationships in the 141 accessions which were benefit for the variety improvement and germplasm resource conservation.