Drought is one of the most important types of abiotic stress that affects stability and amount of yield. This study was conducted to screen for drought tolerance at early seedling stages for 318 ecogeographically diverse wild barley (HordeumvulgareL.spp.spontaneum) diversity collection (WBDC). Considerable variation was observed for all the seedling characters examined. Seedling growth was significantly reduced by 17% polyethyleneglycol -induced osmotic stress with significant variation among accessions. Comparison of mean performance under normal and osmotic stress conditions indicated that shoot length was the most sensitive trait, followed by seedling length, seminal root number, root-shoot length ratio and root length. The mean of root-shoot length ratio, however, increased under osmotic stress. Correlation studies indicated that the root length was the most important trait, followed by shoot length and root-shoot length ratio. The accessions from the Fertile Crescent and North Africa showed more drought tolerance than those from the other geographical regions. The accessions WBDC009 (Jordan), WBDC075 (Libya), WBDC181 (Jordan), WBDC242 (Jordan) and WBDC280 (Israel) exhibited the lowest stress susceptibility index ‘S’ values. Consequently, these accessions showed tolerance to drought at the early seedling stage and are considered to be good sources of drought tolerance for cultivated barley improvement. This work was supported by a grant from Regional Subgenebank Support Program of Rural Development Administration, Republic of Korea .