According to the sequence of 100 ISSR universal primers in UBC801-900 published by Columbia University, 12 materials were selected from 8 species, including Cymbidium goeringii、Cymbidium tortisepalum、Cymbidium goeringii var. longibracteatum、Cymbidium sinense、Cymbidium faberi、Cymbidium kanran、Cymbidium ensifoliumand self-fertility. The results showed that a total of 67 alleles were detected with 11 primers in 96 materials, with an average of 6.09 alleles per locus.The number of Na alleles is 1.9254, the number of effective Ne alleles is 1.4501, the genetic diversity of Nei"s is 0.2767(0.2458-0.3640), and the Shannon index of I is 0.427. The number of polymorphic loci was 62, the percentage of polymorphic loci (PPL) was 92.54%, the genetic diversity (Hs) was 0.1934, the genetic differentiation coefficient (GST) was 0.3009, and the total genetic diversity (HT) 0.2767. Cymbidium has high degree of genetic differentiation and were divided into three groups by systematic clustering anlysis, Cymbidium goeringii and Cymbidium sinense belong to one category, Cymbidium kanran, Cymbidium goeringii var. longibracteatum, Cymbidium faberi, Cymbidium tortisepalum and Cymbidium ensifolium belong to the second category, and the self-fertility is a unique category, which has a large genetic distance from the other two categories. The results of this study can provide an important basis for the breeding and variety identification of cymbidium in the future.