The invention discloses a sainfoin anther callus differentiation culture medium, which comprises KNO3, (NH4)2SO4, Ca(NO3)2, MgSO4*7H2O, KH2PO4, K2HPO4, KI, H3BO3 and other trace elements, iron glycinate, inositol, nicotinic acid, pyridoxine hydrochloride, thiamine hydrochloride, valine, taurine, glutamine, tea polyphenol, ZT, diethylaminoethyl hexanoate, titanium citrate, lanthanum nitrate, polyvinylpyrrolidone, sodium nitroprusside, colchicine, maltose, plant gel and mulberry leaf juice. According to the sainfoin anther callus differentiation culture medium, the green seedling differentiationrate of the sainfoin anther callus is greatly improved, and thus the acquisition rate of regenerated plants from flower culture is improved, and a foundation is laid for the practical application ofthe sainfoin haploid breeding.