Methyl-binding domain proteins (MBDs) are a group of trans-acting factors that bind to methylated DNA sites, and play important roles during the developmental regulation in plants. In this study, ‘Chinese Spring’ was used as materials, and bioinformatic analyses were performed to identify the TaMBD genes in the wheat genome, including composition, sequence characteristics, chromosomal location and expression patterns, Additionally, the spatiotemporal expression patterns of TaMBD6 and TaMBD9 genes were studied by using qRT-PCR technology. The results showed that: (1) 16 TaMBD members (44 gene loci) were included in the wheat genome, and they were distributed on the 1st, 2nd, 5th, 6th and 7th chromosome groups, respectively. Cluster analysis indicated that TaMBDs were sorted into six subclasses Ⅰ, Ⅱ, Ⅲ, Ⅴ, Ⅶ and Ⅷ of eight MBD subclasses in plants. Additionally, 5 conserved binding sites, specific to methylated DNA regions, were found in the TaMBD members of the subclass Ⅱ, Ⅲ and Ⅷ. Gene structure analysis showed that the number of introns is varied at the range of 1 to 10 in TaMBD genes, and the light- and hormone-responsive elements were commonly found in the region of TaMBD promoters. The distribution characteristics of genomic structure and regulatory elements were highly similar among TaMBD members in the same subclass. (2) Gene expression profile analysis of RNA-Seq data showed that most members of TaMBD gene family were highly expressed in grain and spike tissues at the time point of earlier development stages, and some members were obviously responding to both of drought and heat stresses. (3) The qRT-PCR results showed that the 3 homologous genes of TaMBD6 and TaMBD9 were differentially expressed in different tissues, but they were all expressed at the highest level in young spike tissues. The results indicated that the MBD genes might play important regulatory roles during the process of developmental regulation and abiotic stress responding in wheat.