In vitro flowering and embryo rescue are common techniques for improving safflower hybrid fertility, even though anther culture speeds up the generation of superior types. The aim of the study was to evaluate the effect of plant hormones and genotypes on anther culture responses in order to develop a suitable protocol for anther culture. Callus, shoot, and root formation of the local and Turkan safflower genotypes were examined in ten factorial hormone treatments with three replications. On the MS medium, different doses of Thidiazuron (TDZ) at 1 mg/l indole butyric acid (IBA) and variable benzylaminopurine (BAP) at 0.5 mg/l naphthalene acetic acid (NAA) were used for callus induction and shoot regeneration, respectively. Half-strength MS media with different IBA concentrations at a constant 0.5 mg/l NAA were used for root regeneration. Of the two genotypes, Turkan was superior in all aspects of callus induction, while the local safflower performed better in shoot and root regeneration. The average callus generation for both genotypes was 38 and 39% at 0.25 and 0.50 mg/l TDZ; 17% at 2.0 mg/l BAP, and 18% at 1.0 mg/l IBA for shoot and root formation, respectively. For genotype-hormone interaction, the Turkan genotype showed a significant degree of callus formation (46%) at 0.5 mg/l TDZ. Similarly, the local genotype had the maximum shoot (20%) and root (21%) at 2.0 mg/l BAP and 1.0 mg/l IBA phytohormonal treatment, respectively. After being acclimated, the plantlets underperformed in the growth chamber, and none of them survived in the glasshouse

Keywords: Callus induction, Genotypes, phytohormones, Haploids, Regeneration,, Safflower