Hordeum californicum ( H. californicum, 2n=2X=14, H cH c), one of the wild relatives of wheat ( Triticum aestivum L.), harbors many desirable genes and is a potential genetic resource for wheat improvement. In this study, an elite line ND646 was selected from a BC 4F 5 population, which was developed using 60Co-γ irradiated wheat- H. californicum disomic addition line WJ28-1 (DA6H c) as the donor parent and Ningchun 4 as the recurrent parent. ND646 was identified as a novel wheat- H. californicum 6H cS/6BL translocation line using genomic in situ hybridization (GISH), fluorescence in situ hybridization (FISH), and H. californicum -specific expressed sequence tag (EST) markers. Further evaluation revealed that ND646 had excellent performance in several traits, such as a higher sedimentation value (SV), higher water absorption rate (WAR), and higher hardness index (HI). More importantly, it had more kernels per spike (KPS), a higher grain yields (GY), and good resistance to powdery mildew, leaf rust, and 2,4-D butylate (2,4-D). Its excellent phenotypic performance laid the foundation for further investigation of its genetic architecture and makes ND646 a useful germplasm resource for wheat breeding.