X1-homologous genes (XHS) encode plant specific proteins containing three basic domains (XH, XS, zf-XS). In spite of their physiological importance, systematic analyses of ZmXHS genes have not yet been explored. In this study, we isolated and characterized ten ZmXHS genes in a whole-of-genome analysis of the maize genome. A total of ten members of this family were identified in maize genome. The ten ZmXHS genes were distributed on seven maize chromosomes. Multiple alignment and motif display results revealed that most ZmXHS proteins share all the three conserved domains. Putative cis-elements involved in abiotic stress responsive, phytohormone, pollen-specific and quantitative, seed development and germination, light and circadian rhythms regulation, Ca2+-responsive, root hair cell-specific, and CO2-responsive transcriptional activation were observed in the promoters of ZmXHS genes. Yeast hybrid assay revealed that the XH domain of ZmXHS5 was necessary for interaction with itself and ZmXHS2. Microarray data showed that the ZmXHS genes had tissue-specific expression patterns in the maize developmental steps and biotic stresses response. Quantitative real-time PCR analysis results indicated that, except ZmXHS9, the other nine ZmXHS genes were induced in the seedling leaves by at least one of the four abiotic stresses applied.