Abstract:6 SSR markers ralated to traits of diastatic power and enzyme extraction of malting quality were applied to analyze genetic diversity in one hundred and three accessions of wild barley (Hordeum vulgare L.) from Guizhou. As shown in the present study, a total of 38 alleles were detected with an average of 6.33 per SSR primer pair. Our results indicated that locus GMS001 was effectively applied to detect DNA variation among Guizhou wild barleys, based on higher values of polymorphism information content(PIC) as compared with those of Australian barley cultivars. The dendrogram by UPGMA cluster analysis revealed that the selected SSR markers may be applied to separate phenotype and resource classification of 103 materials obtained from Guizhou. Namely, genetic diversity of the wild barley obtained from Zunyi district was abundant, as compared with materials from the other districts. Moreover, more significant genetic difference was observed in six-row wild barley than in -row material. It is suggested that choice of beer barley parents should be performed among populations of six-rowed wild barley in Guizhou, based on their abundant genetic diversity.