Abstract: Three pistils is a mutation in common wheat，it could potentionally increase the number of gains per spike, so it is a valuable mutant for wheat breeding. To study the molecular mechanism of the three pistils mutation in wheat we cloned and characterized the cDNA sequence of TPUCR. Then, the acquired sequence and the amino acids were analyzed and their sequences alignment, secondary structure and functional domains were predicted. Moreover, the TPURD gene was over-expressed in E. coli. The expression patterns of TPURDwere analyzed through real-time quantitative RT-PCR in different developmental stages of young spikes. The open reading frame (ORF) of TPUCR was 381 bp, and it encodes 126 amino acids. Alignment analysis indicates that the deduced amino acid sequence share a high homology with those of other several grain crops. Functional domains analysis indicated that the TPURD contained a conserved domain. Expression analysis indicates that there are higher levels of transcripts in the earlier developmental stages of young spikes. The transcript levels in the young spikes of Chinese Spring three pistils line increased by 4-fold compared with that in the young spikes of the Chinese Spring line. The data provide a basis for further research on function of TPURD, and a rationale for the molecular mechanism of the three pistil mutation in wheat