Abstract: The anthers in uninucleate stage were inoculated and plantlets of three cultivars such as Surprise des haues, Redgauntlet and Harukaoli were regenerated directly from the callus formed on MS medium supplemented with 4 ppm LAA, 2 ppm K and 2 ppm BA. Plantlets of Sofia, Hokowasei, Gorella. Redcoat were differentiated directly from the callus formed on MS medium in which the component and concentration of hormones were changed. The four main strawberry viruses including SMoV, SMYEV, SVBV and SCrA were not detected in the anther plantlets of Surprise des haues, Redgautlet, Harukaoli, Hokowasei and Sofia by means of leaf grafting method. The growth and yield of virus-free plants of four cultivars wese considerably different from the control cultivars.