Abstract:Gui culture medium has been developed by adjusting the concentration of macro- and micro- elements, based on the studies of various media applying to wheat ( Triticum aestivum L. ) anther culture breeding for many years. An appropriate ration among NH4N03, KN03, MgSO4 and MnSO4 in the medium was selected by orthogonal design L16 (45). For comparison of different media namely C17, W14 and N6 medium, the average frequencies of callus induction were increased by 30.31%, 50.66% and 57.96% respectively when Gui medium was used. And the Gui medium showed all its advantages in various crosses. The frequencies of wheat anther callus induction showed an increase of 54.25 % to 64.07 % and the callus development was also promoted when adding 0.9 mg/L rare earth addition to the Gui medium.