The level of β-glucan which is a major soluble dietary fiber found in the grain endosperm cell wall was highly variable among 25 barley genotypes grown at four locations including Suwon, Naju, Jinju, and Jeju. Statistically significant genotypic effects were observed for β-glucan content at each or across growing sites (P<0.001). On average, 'Chalssalbori' showed the lowest percentage β-glucan (4.04%) among genotypes in the grain, whereas 'Yonezawa Mochi' was highest in percentage β-glucan (6.46%) compared to other genotypes. The significant difference between genotypes was approximately 1-2% across environments. The effects of location or interaction between locations and genotypes were not significant on the variation of β-glucan contents. High β-glucan content seemed to be greatly associated with such grain traits as waxiness and presence of husk except for 'Chalssalbori'. The waxy genotypes had a mean of 5.37% and values ranging from 5.28 to 5.47%, but normal genotypes had a mean of 4.78% and values ranging from 4.69 to 4.88% over environments. Hulless barley genotypes were also higher than hulled barley genotypes for the average β-glucan content in both individual and over all environments. The difference between the hulled and hulless gene pools was on average of 0.37% with ranges from 0.19% to 0.56% at four environments. β-glucan content measured from a mapping population of F5 -derived 107 lines derived from the cross between 'Yonezawa Mochi' and 'Neulssalbori' was not significantly associated with other agronomic traits except for 1,000-kernel weight at the '01 Suwon environment. Not too much information on the relationship of β-glucan content to agronomic traits was available.