Genetic diversity was analysed in 52 Argentinian populations of Avena barbata, a tetraploid grass introduced in America from Spain during the colonization period. Nine isozyme systems were studied and 14 loci identified, five of which were polymorphic. Cluster analysis based on Hedrick's index revealed a high similarity among populations. The total diversity (Pt) in the 52 populations was 0.144, the mean diversity (Ps) was 0.04, while between population diversity (Dst) was 0.103. The resulting coefficient of differentiation (Gst) was 0.714, indicating that diversity among populations was an important contributor to the total variability. Genetic diversity was structured into multilocus associations; 122 different complexes were found among 3311 individuals, but only two complexes occurred at a high frequency. The distribution pattern of these frequent multilocus genotypes was associated with environmental factors, mainly rainfall and temperature. The comparison of these results with those of previous studies on A. barbata from Spain indicated that Spanish and Argentinian populations are closely similar in allelic composition on a locus-by-locus basis but different in multilocus genotypic composition. We concluded that selection was the main force involved in the reorganization of the Spanish genepool into novel multilocus associations adapted to specific habitats in Argentina.