Carduus acanthoides L (spiny plumeless thistle) a biannual wild species with 2 n = 22 chromosomes was crossed with Helianthus annuus L. When crossing pollen from C . acanthoides germinated and pollinated the stigmas of sunflower lines HA 89A after 48 h. The crossability rate was low but seeds and hybrid plants were obtained. The F 1 plants strongly resembled the cultivated sunflower with the most important bio-morphological characters even though they had an intermediate type of heritability. The hybrid nature was confirmed by RAPD markers. The polymorphism between H . annuus C . acanthoides and their F 1 hybrids was studied using RAPD. The result showed introgression of C . acanthoides in the hybrid progeny. It was established that the wild species carried Rf genes for the CMS PET-1. After self-pollination and sib-pollination of the F 1 plants and back-crossing with cultivated sunflower F 2 BC 1 and next generation hybrid progenies were obtained. The investigation encompassed the period 2000–2007 and 2014–2018. Some of the new lines have been included in a heterosis breeding program for developing hybrids for the sunflower market.