Abstract:The WRKY proteins are broadly investigated in plants in the past twenty years. Presently, many members of this gene family have been identified and characterized in more than 10 plant species. WRKY genes were also isolated from wheat, but functional characterization of these genes is largely restricted by the lack of bioinformatic analysis. In this study we cloned the TaWRKY74-c and TaWRKY74-d genes, which shared high similarity with TaWRKY74 genes in public GenBank database and encoded transcription factors with 344 and 371 amino acids, respectively. These proteins contain highly conserved WRKY domain and belongto Class III of WRKY transcription factors family. Real-time PCR indicated that target gene expressed in flower, leaves and stem with the highest level in stem and the lowest in flower. In order to comprehensively clarify the genes functions, Genevestigator tools were used to detect genes expression patterns under 331 environmental stimuli, 10 developmental stages and 21 tissues or organs. Results showed that genes played important roles throughout plant development stages and in plant responses to cold acclimation and pathogens. We then carried out RT-PCR to verify these results. The data indicated that the expression patterns of TaWRKY74 gene under different conditions are highly consistent with bioinformatic predictions. Bioinformatic tools with some recently developed programmes in this study enriched our knowledge in wheat WRKY gene functions and provided crucial clues and information for further clarification of these genes.