Abstract: β-glucan content spatial distribution characteristics of cultivated barley in Qinghai-Tibet Plateau were investigated based on the data measured from 83 sampling sites. The results showed that β-glucan content exhibited a macrocosm pattern of spatial distribution along the horizontal direction in Qinghai-Tibet Plateau, with staggered patches of different values. Two regions of higher β-glucan content were observed with centers of Guide and Tongde in Qinghai Province and Xiahe and Hezuo in Gansu Province (northeast), and Jiangzi, Bailang, Xietongmen, Shigatse, Lazi, Kangma, Gongga and Qushui in Tibet (southwest). Along the vertical direction, β-glucan content distribution pattern appeared with double peak curves. There were two high value zones between the altitudes from 2700 m to 3000 m and from 3600 m to 3900 m, with average β-glucan content value of (5.7±1.7)% and (4.6±1.1)%, respectively. Influencing factors of β-glucan content with importance value index more than 40.0% were as follows: grain color ＞ ear density ＞ average relative humidity in September ＞ soil available N content ＞ soil available K content ＞ average diurnal temperature range in June ＞ ≥10 ℃ accumulated temperature ＞ average annual temperature ＞ soil available P content ＞ average diurnal temperature range in September.