Abstract: Wheat sterile mainly expressed through the performance of pollen abortion. The study on infertility material of wheat heterosis has great significance. Overseas research shows that among certain parts of the common wheat hybrids F₁, pollen sterility is controlled by nuclear genome of pollen killing gene Ki. In order to screen linked markers of pollen killing gene Ki in wheat, modern molecular biology techniques were used to locate the gene, clone fragment pollen lethal gene linked markers, provide an effective selection marker for transformation of male sterility in wheat germplasm and study the molecular markers of pollen killing gene Ki in wheat. A BC₁F₁ population derived from the crossing between Chinese spring wheat and Australian spring wheat was used for molecular mapping of Ki gene. The sterile and fertile near isogenic bulks was constructed from the above BC₁F₁ population. By using Bulked Segregant Analysis (BSA) method, 85 SSR primers on wheat 6B chromosome were screened, and two SSR markers (Xgwm626 and Xgpw4138) were found to be linked to Ki gene. These markers could be used in fine mapping of Ki and marker-assisted selection. Two SSR markers Xgwm626 and Xgpw4138 located on the long arm of chromosome 6B were linked to Ki, and their genetic distances to Ki were 9.2 cM and 6.9 cM, respectively, indicating that these two markers could be used in molecular-assisted breeding.