To study the relationship between different Saccharum spontaneum L. genotype clones, karyotype analysis of these clones were performed by using the method of wall degradation and low osmosis. The results showed that most of the chromosomes were median region (m) types, only a few were submedian region (sm), median point (M), terminal point (T) and terminal region (t) types in all of the materials used for study. The karyotype formulas of each accession were summarized based on the datum. The karyotype of 2 accessions exhibited 1B type, 7 accessions for 2B and 1 accession for 2C. All of the materials showed karyotypic difference and asymmetry types.