Sorghum-Sudan grass which is an annual forage grass in Gramineae have been bred for biomass promotion. The recombinant inbred line (RIL) populations were derived from the F2:3 genetic mapping population of Sorghum 314A×zongke Sudan grass. The breeding selection based on agronomic characters and DNA molecular marker statistical analysis was carried among the progenies of the RIL populations, and 9 recombinant breeding progenies were determined as heterobeltiosis with high yield. According to DNA diversity analysis of simple sequence of repeats(SSR) marker, the genetic varieties were abundant among the families, and the recombinant breeds of high yield were distributed among the different RIL populations.2 family lines, which have similar agronomic characters and yield, and whose genetic distance nearly equal to zero, showed difference on arid tolerance. 4 breeding lines were selected on agronomic characters and resistance in the field trials, and 2 breed lines were qualified for the regional testing.