Abstract: Genetic diversity is essential for genetic improvement of a given crops. If the information on genetic diversity is not enough to utilize, the available genetic diversity study should be crucial. The objective of this study was to estimate the variability, heritability and genetic advance of barley landraces collection from Gamo highlands of Ethiopia. Accordingly, thirty six barley landraces were evaluated at Chencha, Southern Ethiopia in 2014/2015 cropping season to obtain information on genetic variability, heritability and genetic advance for grain yield and 12 yield related characters in Ethiopian barley landraces. The plot design used for the experiment was a randomized complete block design with three replications. The analysis of variance for the 36 barley landraces revealed significant difference among the landraces for the 13 quantitative characters studied, except spike length. The phenotypic coefficients of variability were higher than genotypic coefficients of variability for all of the characters indicating each and every characters were influenced by environmental factors up to some extent. The greater difference between GCV and PCV was observed spike and peduncle length indicating that these characters were influenced by environmental factors to greater extent. Heritability coupled with high genetic advance was observed for characters biomass per plant, grain yield and number of tiller per plant indicating that selection for these characters could be more effective due to additive gene action. Thus, this study revealed the presence of sufficient variability among the barley landraces in the country that can be exploited for germplasm enhancement.