The diversity within 16 natural progenies of the grapevine dieback fungus, Eutypa lata, was investigated by sampling single-ascospore isolates mainly in France and using random amplified polymorphic DNA (RAPD) markers, vegetative compatibility (VC), and pathogenicity testing. The combination of RAPD and VC data identified each isolate as a unique genotype within each progeny. Only three RAPD haplotypes did not cluster within the expected groups, i.e. the ascospore families. Within each set of clustering haplotypes, Mendelian 1:1 ratios for absence and presence were observed for RAPD markers, indicating that each progeny was the result of a biparental cross. Only one mycelium was obtained when isolation was performed from the discolored wood sustaining the perithecial stroma. This mycelium was identified as a likely parent of the corresponding progeny by RAPD analysis. The level of diversity measured by the average distance between haplotypes calculated from RAPD data, the percentage of vegetatively compatible pairs and the range of pathogenicity appeared similar between all but one progeny, indicating that crosses occurred within a random-mating population. All the results were consistent with the hypothesis that E. lata is a random-mating species having a high degree of genetic diversity.