Hordeum chilense is a wild barley species that has a high degree of genetic variability and significant potential for use in plant breeding. To establish a series of trisomics in H. chilense (2n = 14), plants with 2n + 1 chromosome numbers were isolated from the progenies of selfed triploid plants. Based on both fluorescent in situ hybridization with pAs1 and pTa71 repetitive DNA probes and C-banding patterns, seven different trisomics were tentatively identified. Primary trisomic plants were for chromosomes 1Hch, 4Hch, 5Hch, 6Hch and 7Hch. A secondary trisomic carrying a 5HchS-5HchS isochromosome as the extra chromosome and a trisomic for chromosome 3Hch heterozygous for the 3HchS-4HchL and 4HchS–3HchL interchange were identified. The trisomic for chromosome 1Hch cannot be phenotypically distinguished from the diploid. The rest of the trisomic types were distinguishable from the diploid by their morphological characteristics (relatively poor vigour, decreased size and shorter spikes) but they were morphologically indistinguishable from each other. The frequencies of trisomics among the progenies derived from self-fertilization of these aneuploids ranged from 10.7% to 37.5%, with an average frequency of 26.1%.