Improvement of yield-traits is one of the predominating objectives in wheat breeding. Homeodomain-leucine zipper (HD-ZIP) transcription factor plays significant roles in plant growth and development. The TaHDZ34 (A, B and D sub-genomics) genes consisting of three members of the HD-ZIP IV transcription factor gene subfamily in wheat (Triticum aestivum L.) were cloned. Two haplotypes of TaHDZ34-7A, TaHDZ34-7B or TaHDZ34-7D were respectively identified after the sequence polymorphism analysis, and three functional molecular markers were developed. The TaHDZ34 genes were divided into eight haplotype combinations. Association analysis and distinct population validation jointly indicated that TaHDZ34 had the function of modulating grain number per spike, effective spikelet number per spike, 1,000 kernel weight, and flag leaf area per plant in wheat. Among all haplotype combinations of TaHDZ34, Hap-ABD was the most excellent one. Subcelluar localization showed that TaHDZ34-7A was localized in the nucleus. Interaction proteins of TaHDZ34-7A protein proved to be involved in protein synthesis/degradation, energy production and transportation, and photosynthesis processes. Geographic distribution and frequencies of TaHDZ34 haplotype combinations suggested that the Hap-Abd and Hap-AbD were preferential selection in Chinese wheat breeding programs. The high-yield related haplotype combinations Hap-ABD of TaHDZ34 provided beneficial genetic resources for marker-assisted selection of new wheat cultivars.