Abstract: Grain protein content of 287 barley germplasms from different origins were analyzed by using near infrared method in this study to understand the property of protein content and extend parent resources. Distribution and differences of grain protein content among diverse origins were compared and the results showed, grain protein content of 287 barley accessions ranged from 8.00% to 20.10%, averaged 12.43%, and assembled around the scope between 10.00% and 14.00%. The minimum protein content of accessions, 8.00%, derived from Yunnan Province, while the maximum, 20.10%, from Qinghai. Significant divergence of protein content existed among origins with the highest value of 14.49% arisen in Henan Province and the lowest value of 9.62% in Inner Mongolia. The order of origins based on protein content from high to low was Henan, Qinghai, Xinjiang, Jiangsu, Hubei, Yunnan, Anhui and Inner Mongolia. Variation of barley grain content among different origins reflected its diverse uses.