Miscanthus is a C₄ herbaceous perennial genus, and it was chosen as a bioenergy crop due to high biomass yield. Miscanthus sinensis has many phenotypes which are adapted to various environments in China. In this study, 421 accessions of M. sinensis were collected from 22 provinces, and the genetic polymorphisms amongst these germplasm collections were identified using 20 primer pairs designed against 10 each from expressed sequence tag-simple sequence repeats [EST-SSR (eSSR)] and genomic SSR (gSSR) transferable markers from barley. A total of 95 SSR polymorphic bands were detected producing a 100% polymorphic rate among these M. sinensis accessions. The gSSR markers showed a richer genetic polymorphism than eSSR markers. Based on the unweighted pair group method with arithmetic mean (UPGMA) clustering, there was a distinct sub-population separation in M. sinensis, which indicates that geographical differences and natural selection are the driving forces for genetic variation and evolution in the species. The 20 pairs of barley markers matched to 26 polymorphic bands associated with date of heading, plant height, leaf weight, stem weight, leaf/stem ratio, and total biomass yield. Eleven marker polymorphic bands were associated with the date of heading, 4 with plant height, 10 with leaf weight, 7 with stem weight, 3 with leaf/stem ratio, and 10 with biomass yield.