Sorghum is the fifth important crop in the world. It is also the major food resource in African countries. Striga asiatica is a parasitism weed on sorghum and some other important crops. In this report, two sorghum lines with the difference in response to Striga asiatica, SRN39 (lower Germination Stimulant-GermStim production) and Shanguihong (high GermStim production), were selected as the parents for the construction of a recombinant inbred (RI) population. Ninety-four RI lines were collected for the molecular analysis and GermStim production evaluation. A genetic map was constructed with 251 molecular markers that distributed on 10 different linkage groups. The map covers sorghum genome of 1,779 cm with an average map distance of 7.1 cm between linked markers. It is one of the complete sorghum molecular map in the world. Co-segregation analysis indicated that the germination stimulant gene (GermStim) was located on linkage group J, which was at a distance of 13 cm from the closed marker. Further RAPD analysis between two parents and two DNA pools different in the amount of germination stimulant production, several polymorphic DNA fragments were identified and cloned. Mapping results showed two of them flanked with the GermStim gene at a distance of 1.6 cm and 2.1 cm respectively.