The isonucleus-allocytoplasmic materials ms(S)-Xinong 1376, (A)-Xinong 1376, and ms(A)-Xinong 1376 were used to investigate the protein expression profile in mononucleated anaphase anthers of wheat (Triticum aestivum L.) after silver staining and analysis by PDQuest software. About 320 to 350 distinct protein spots were detected reproducibly. The results indicated that there were some difference in the protein quantity between the two sterile lines and the maintainer line. Sev-eral specific spots were present in the maps of two sterile lines, but were absent in the maintainer line. The two sterile lines had their own specific expression protein spots with different sterile mechanism. We compared the three maps and con-cluded that the sterile lines may inhibit related protein expression, and turned on some proteins related to male sterility, which disturbs the normal metabolism of substances and energy in anthers and leads to the occurrence of male-sterility.