Populational variations of Echium italicum were analysed based on RAPD, fatty acid and micromorphological patterns of the nutlets. Some minor differences in nutlet morphology and the surface sculpturing were detected. Total oil concentrations ranged from 19.3 [A2(E)] to 23.5 % (A1) among the populations. Significant variations between accessions were detected for fatty acid concentrations. Major unsaturated fatty acids were alpha-linolenic, linoleic, oleic, stearidonic and gamma-linolenic acids respectively. The highest values for stearidonic (15.48 %) and gamma-linolenic acid (7.66 %) were obtained from A2(E) population. The total amplified products of 15 RAPD primers was 220 (average of 14.6 bands per primer), of which 156 bands were polymorphic, corresponding to nearly 70.9 % genetic diversity. The number of bands for each RAPD primer varied from 7 (OPD14) to 30 (OPC6). Percent of polymorphic bands ranged from 0.7 % (OPC19) to 100 % (OPC4 and OPD19). Examined traits showing high polymorphism seems to be useful for the taxonomical circumscription, infraspecific delineations and segregation of the populations of E. italicum. Based on the dendrograms of RAPD and fatty acid data, investigated accessions may be separated into subgroups A1, C2, and Irano-Turanian and Mediterranean. Obtained results may be also evaluated for characterization of E. italicum germplasm in selection of high yield genotypes for the production of long chain omega-3 PUFAs including SDA and GLA, and improvement of oil crops in the field.