Aim of the study was to investigate the effect of long-term fertilization on the quality of wheat grain including protein, starch, fat content and the amino acid was analyzed by cluster analysis. The effect of fertilization on the quality of wheat seed was investigated based on continues application of fertilizer for 26 and 36 years in Gleyic Cambisols. The results showed that the amounts of protein were 14.12% and 14.1% for N_2 and N_2PK treatments which was higher than others. In addition, the total amounts of amino acid decreased by fertilization. However, the amounts of starch decreased and the less decreased treatment was N_2PK treatment for 0.07%. The highest fat was 0.17% for N_2PK treatment.