We investigated the allelic nature and map locations of Hordeum vulgare (barley) homologs to three classes of Arabidopsis low temperature (LT) regulatory genes-CBFs, ICE1, and ZAT12-to determine if there were any candidates for winterhardiness-related quantitative trait loci (QTL). We phenotyped the Dicktoo x Morex (DxM) mapping population under controlled freezing conditions and in addition to the previously reported 5H-L Fr-H1 QTL, observed three additional LT tolerance QTLs on 1H-L, 4H-S, and 4H-L. We identified and assigned either linkage map or chromosome locations to 1 ICE1 homolog, 2 ZAT12 homologs, and 17 of 20 CBF homologs. Twelve of the CBF genes were located on 5H-L and the 11 with assigned linkage map positions formed 2 tandem clusters on 5H-L. A subset of these CBF genes was confirmed to be physically linked, validating the map position clustering. The tandem CBF clusters are not candidates for the DxM LT tolerance Fr-H1 QTL, as they are approximately 30 cM distal to the QTL peak. No LT tolerance QTL was detected in conjunction with the CBF gene clusters in Dicktoo x Morex. However, comparative mapping using common markers and BIN positions established the CBF clusters are coincident with reported Triticeae LT tolerance and COR gene accumulation QTLs and suggest one or more of the CBF genes may be candidates for Fr-H2 in some germplasm combinations. These results suggest members of the CBF gene family may function as components of winter-hardiness in the Triticeae and underscore both the importance of extending results from model systems to economically important crop species and in viewing QTL mapping results in the context of multiple germplasm combinations.