MADS-box genes interact with TB1 to regulate plant organ morphogenesis. In rice, OsMADS57 interacts with OsTB1 to control OsD14 transcription. In this study, we aimed to determine the relationships among these genes in barley. We identified a natural mutant of HvTB1 (HvTB1x) formed by a C→A transition at position 230, which resulted in a premature stop codon. We cloned the HvMADS57 and HvD14 genes and studied their expression in the tb1 mutant. The results showed that tHvMADS57 is a MIKCc-type MADS-box gene, and the expression levels of both HvMADS57 and HvD14 were significantly reduced in the tb1 mutant when compared to those in the wildtype gene. These results indicate that, HvMADS57 regulates plant growth and development by interacting with HvTB1 to suppress the transcription of HvD14 in barley which is similar to the relationships among the orthologs of these genes in rice.