Cultivated barley (Hordeum vulgare L. ssp. vulgare) is one of oldest domesticated crops and four most important cereal species. In addition to end-use for animal feeding, malting and human diets using barley grains, the straw of cultivated barley serves as a silage grass for animals in some parts of barley cultivation areas (e.g. Tibet). In order to identify elite barley resources in future breeding for silage barley varieties, this study analyzed the phenotypic variation 102 barley germplasm accessions from National Crop Genebank of China in Chongqing, south-west part of China. We determined the phenotypic variation on 15 traits and conducted statistical analysis using Shannon-Weaver Index, correlation analysis, cluster analysis and PCA analysis. These results suggested that the Shannon-Weaver Index (H’) ranged from 0.729 to 0.957. The higher phenotypic variation was detected on leaf yellowing at heading and filling stages, plant lodging and 1000-grain weight, but the narrowed diversity was found on grain width and plant height. In addition, we found that the phenotypic diversity of winter barley ecological region (0.721) was slightly higher than that of southwest and Tibet Plateau region (0.706). Four subpopulations were clarified by clustering analysis, being consistent with the geographic origins of tested materials. Moreover, a number of barley germplasm resources, which showed high plant height, lodging resistance, large leaves, staying green of old leaves, were identified. These elite barley germplasms might become useful potentially in breeding for silage barley varieties in areas of Chongqing, China.