TILLING (Targeting Induced Local Lesions IN Genomes) is a reverse genetics method which identifies individuals carrying allelic variants at a gene of interest, within a chemically mutagenized population. TILLING has a number of positive features, including the capacity to provide the investigator with single point mutations offering a wide range of genetic effects useful in functional studies or in breeding applications. Additionally, mutant lines are not regulated as genetically modified. In barley, TILLING has been applied since 2004 and at least six TILLING resources have been described. Mutation densities ranged from 1 per 154 to 1 per 2,500 kb, suggesting that efficient barley TILLING requires a few thousands mutagenized plants. In this review, we will address the main aspects and results of barley TILLING applications. We will give special emphasis on technical novelties in molecular screening approaches, and on rationale and opportunities of using TILLING in barley breeding.