In order to understand the genetic diversity of Cymbidium tortisepulum, the fingerprints of 32 C. tortisepulum cultivars were constructed by SSR. The results showed that there were 95 alleles were detected by selected 12 pairs of primers, the number of alleles selected by one primers ranged from 4 to 18, and the number of mean alleles (N_A) and effective alleles (N_E) were 61.489 and 5.124, respectively. The Shannon information index (I) and polymorphism information content values (PIC) of 32 cultivars ranged from 0.806 to 2.624, and 0.789 to 0.953, respectively. Among 12 paris of primer, the primer SSR03 was the highest in N_A, N_E, observed heterozygosity (H_O), I and PIC. All of 32 cultivars had different specific bands by 12 pairs of primer. The SSR molecular fingerprints of 32 cultivars were built by 3 core pairs of primers, including SSR02, SSR03 and SSR12. All of 32 cultivars could be distinguished each other by combined primers of SSR02 + SSR03 + SSR123. These would provide theoretical basis and technical support for studying in the identification of C. tortisepalum cultivars, genetic diversity analysis and molecular breeding.