To understand genetic structure and diversity of parental cultivars involved in China Mainland sugarcane breeding programs,92 elite parents and 4 wild relatives were genotyped with 18 microsatellite DNA markers.The genetic similarity(GS) values among the cultivars ranged from 0.346 to 0.960 with an average of 0.533.Among the introduced cultivars,India accessions had the closest genetic distance to China Mainland accessions(0.447),while Australia accessions have the furthest distance(0.503).A comparison of allelic diversity among geographical origins showed that there were 22 China Mainland specific alleles,of which 28% were derived from native S.spontaneaum germplasm in China.Model-based genetic structure,clustering,and principal components analyses consistently revealed there were five groups within the 96 accessions.Groups 1,2,4,and 5 consisted of all cultivars and group 3 only contained wild germplasm.Group 2 was characterized as the Introduction group with 46 cultivars predominantly introduced from Australia,Taiwan of China,India,and USA.Groups 1,4,and 5 consisted of cultivars mostly originated from China Mainland,defined as the Complex group,Yacheng lines group,and F134/CP72-1210 group,respectively,upon their pedigree.By understanding the genetic relationships among the parental cultivars,breeders can gain a rational basis for expanding the gene pool and select the best parental accessions for crossing. To understand genetic structure and diversity of parental cultivars involved in China Mainland sugarcane breeding programs, 92 elite parents and 4 wild relatives were genotyped with 18 microsatellite DNA markers. Genetic similarity (GS) values ​​among the cultivars ranged from 0.346 to 0.960 with an average of 0.533.Among the cultivars, India accessions had the closest genetic distance to China Mainland accessions (0.447), while Australia accessions have the furthest distance (0.503) .A comparison of allelic diversity among geographical origins showed that there 22 alleles, of which 28% were derived from native S.spontaneaum germplasm in China. Model-based genetic structure, clustering, and principal components analyzes who found there were five groups within the 96 accessions. Groups 1,2,4, and 5 consisted of all cultivars and group 3 only contained wild germplasm. Group 2 was characterized as the Introduction group with 46 cultivars predominantl y introduced from Australia, Taiwan of China, India, and USA. Group 1, 4, and 5 consisted of cultivars mostly originated from China Mainland, defined as the Complex group, Yacheng lines group, and F134 / CP72-1210 group, respectively, upon their pedigree.By understanding the genetic relationships among the parental cultivars, breeders can gain a rational basis for expanding the gene pool and select the best parental accessions for crossing.