[Objective] To evaluate the genetic background of triploid female and diploid male of Siraitia grosvenorii and to provide biological references for fine varieties breeding of seedless S. grosvenorii. [Methods]Inter simple sequence repeats(ISSR) marker was developed to analyze the genetic background among 28 samples of S. grosvenorii,and cluster analysis and double principal coordinate analysis were revealed by the NTSYS-pc software and Gen AIEx software,respectively. [Results]13 ISSR primers selected out of 100 primers for amplification,and a total of 131 unambiguous bands were obtained,among which 99(PPB = 75. 57%) were polymorphic. The results of cluster analysis and double principal coordinate analysis showed that there was a certain richness of genetic background in triploid female and diploid male of S. grosvenorii. But the genetic similarity coefficients of majority were relatively great with close genetic distance. [Conclusions] The triploid female and diploid male of S. grosvenorii had relatively low complexity of genetic background,so that germplasm innovation strategies should be carried out to enrich the genetic background of seedless S. grosvenorii. [Objective] To evaluate the genetic background of triploid female and diploid male of Siraitia grosvenorii and to provide biological references for fine varieties breeding of seedless S. grosvenorii. [Methods] Inter simple sequence repeats (ISSR) marker was developed to analyze the genetic background among 28 samples of S. grosvenorii, and cluster analysis and double principal coordinate analysis were revealed by the NTSYS-pc software and Gen AIEx software, respectively. [Results] 13 ISSR primers selected out of 100 primers for amplification, and a total of 131 The results of cluster analysis and double principal coordinate analysis showed that there was a certain richness of genetic background in triploid female and diploid male of S. grosvenorii. But the genetic similarity coefficients of majority of relatively great with close genetic distance. [Conclusions] The triploid female and diploid male of S. grosven orii had relatively low complexity of genetic background, so that germplasm innovation strategies should be carried out to enrich the genetic background of seedless S. grosvenorii.