The goal of this study is to understand the evolution relationship of the members of the B-hordein gene family in hull-less barley by analysis of their structure and to explore their utility in grain quality improvement.Six copies of the B-hordein gene (Hn1-Hn3,Hn7-Hn9) were cloned from six hull-less barley cultivars collected from Qinghai-Tibet Pleateau and molecularly characterized.Comparison of their predicted polypeptide sequences with the published data suggested that they all share the same basic protein structures.In addition,we found that the C-terminal end sequences of all B-hordeins shared a similar feature.In the six clones and the other three published genes (Hn4,Hn5,Hn6) from hull-less barley, Hn2 and Hn7 contained the identical C-terminal end sequence DIMPVDFWH. Hn3,Hn4,Hn5,Hn8 and Hn9 also shared the common sequence DIMPPDFWH, which was similar to that of a B-hordein reported previously. Both Hn1 and Hn6 exhibited differences in their C-terminal end sequences,and they clustered into different subgroups.The B-hordeins with the identical C-terminal end sequences were clustered into the same subgroup, so we believe that B-hordein gene subfamilies possiblely can be classified on the basis of the conserved C-terminal end sequences of predicted polypeptide. Phyloginetic analysis also indicated that there is a relatively weak identity between our predicted B-hordein and those reported from H.chilense and H.brevisubulatum. All of our nine predicted B-hordeins were clustered together and other B-hordeins formed another cluster.The possible use of these genes in relation to barley quality is discussed.