Monitoring and protecting germplasm in genebanks using in situ collections while preserving its original genetic integrity is a priority of germplasm curation. Many germplasm accessions need to be regenerated due to their demand and/or seed condition. The regeneration of wild Helianthus (sunflower) species poses several challenges due to the diversity of 53 wild species. Fatty acid composition of sunflower oil is an important quality factor for the crop. Since oil quality is environmentally influenced, and evaluation of this trait is usually performed on oil from achenes from the original accessions of wild sunflower species, we conducted a study on 72 accessions of eight annual and four perennial taxa of wild sunflower species to compare the oil quality of the original accessions and those regenerated for genebank maintenance. The results showed that the fatty acid composition profiles of achenes from the original and regenerated accessions are not the same. It seems that selection for specific fatty acids in several species will require the analysis of both populations to identify germplasm accessions for use in breeding programmes. It should be borne in mind that accessions of wild species are open-pollinated segregating populations, so one would expect some variability in each succeeding generation. While there may be differences between the original and regenerated accessions, the interrelationships of fatty acids are generally similar in wild and cultivated sunflower species, so there should be no detrimental effects on oil quality when using the wild species for other traits. As more regenerated accessions become available, a more precise relationship between the original and regenerated accessions should emerge.