Safflower, Carthamus tinctorius L. (2n = 2x = 24), is one of the world’s oldest oilseed crop with numerous past and present uses. Similar to the main crops, producibility limitations could be overcome by broadening the genetic base of breeding materials through interspecific hybridization. Natural and artificial hybridization between safflower and several wild relatives have been reported. The wild species of C. oxyacantha Bieb (syn. C. oxyacanthus ) (2n=2x=24), can be simply exploited to broaden the genetic base of safflower through interspecific hybridization. This species has comparable characteristics including agronomic traits, oil content and fatty acid composition and even resistance to safflower fly to cultivated safflower? however, there is lack of information on utilization of this species to enrich safflower genepool. Over the last few decades, molecular markers, such as AFLP (amplified fragment lengths polymorphism), ISSR (inter simple sequence repeat) and RAPDs (random amplified polymorphic DNA) have become so important for generation of genetic linkage maps. These DNA-based markers in safflower have just been applied to assess interaspecific genetic diversity among safflower genotypes and reclassification of the genus Carthanus L. Intersimple sequence repeats (ISSR) is a PCRbased marker in which 1625 bp long microsatellites prime in a single primer PCR reaction to amplify the intermicrosatellite regions. This technique has been extensively employed to estimate interand intraspecific genetic variation in a wide range of crops and recently among cultivated safflower and its wild relatives. In this research, ISSRpolymorphism was employed to study crossabilty of cultivated safflower, C. tinctorius L., and wild relative, C. oxyacantha Bieb., 2) determine the cultivated and wild parentals' genome contribution into F2 progenies and 3) asses the efficiency of transferring parental specific markers in three crosses between two cultivated genotypes, C4110 and C111, and one accession of wild species, ISF2 using 25 ISSR primers. Two spineless, white seeded and red-flowered genotype of cultivated safflower, C4110 and C111, and a spiny, yellow-flowered and brown-seeded accession of C. oxyacantha , ISF2, were used as parents in a paired reciprocal crosses established in 2007. F1 hybrids were detected on the basis of morphological characters and ISSR fingerprinting. Three F2 populations with totally 140 progenies were obtained after intercrossing ten F1 hybrids. Young leaves of parents, hybrids and F2 progenies were sampled after 4 weeks and stored frozen (20 ?C) until DNA extraction.Total genomic DNA was extracted from fully expanded young leaves of parents and F2 individuals according to the Cetyltrimethylammonium Bromide (CTAB) method. A set of 25 ISSR primers capable of generating polymorphic and repeatable bands were used to amplify inter-SSR sequences. Two of used primers w