Production of microspore-derived embryos from cultured anthers is now a well-established technique for the isolation of homozygous lines in sunflower. Experiment was conducted to optimize the culture composition for wild Helianthus and inerspecific hybrids. Anthers of uninucleate microspores were cultured in basal MS media with different hormonal combinations. The induction of callus was quick and proliferation high with the presence of 2,4-D, low cytokinin and auxin contents. Excess cytokinin and auxin had no significance in callus induction. Regeneration potential increased with the increased/optimum amount of casein hydrolysate and benzyl amino purine. Kinetin had no specific influence of regeneration frequency of callus. Genotype had a significant effect on the capacity of callus induction and plantlet formation. H.annuus × H.tuberosus responded well and produced plantlets in optimum culture medium. Androgenesis of interspecific hybrid H.annuus × H.tuberosus formed plantlets that surpassed the embryoid stage.