91 wild barley samples from the Near East，Central Asia and Tibet of China were selected and phylogenetically analyzed based on the alcohol dehydrogenase Ⅲ （ADH3） gene to study the genetic evolution relationship of barley.The results showed that the number of haplotypes （H=22） and genetic diversity （H_d=0.914，π=0.012 65） of the wild barley from Near East were significantly higher than that of the wild barley from Tibet （H=5，H_d=0.753，π=0.012 16） and Central Asia （H=8，H_d=0.810，π=0.011 10）.Ninety-one wild barley samples were mainly divided into two groups （Ⅰ and Ⅱ）.81% of wild barley from the Near East and 80% of wild barley from Tibet were clustered in group Ⅰ. 91% of wild barley from Central Asian and a few parts of wild barley from the Near East and Tibet were clustered in group Ⅱ. The results of analyzing population structure further confirmed that the wild barley samples from Tibet were more closely related to wild barley from the Near East than to wild barley from the Central Asia.It is clarified the genetic evolution relationship of wild barley in the Near East，Tibet and Central Asia.It will provide the molecular basis for studying the origin of cultivated barley.