Spectral reflectance and humic substances components were determined aiming at studying the effects of different fertilization on humic substances content and color of black soil based on 30 years long-term location trial. A total of 5treatments were included in this study: (1) Fallow; (2) No fertilizer application (CK); (3) Only chemical fertilizer (NPK); (4) Organic fertilizer partly replaced chemical fertilizer (NPKM); (5) Straw partly replaced chemical fertilizer (NPKS). Results indicated that soil organic carbon content in Fallow, NPKM, NPKS were significantly increased by 49.7%, 74.3%, 27.0% in contrast with NPK (p<0.05). The highest content of humic acid (HA) was 3.9 g/kg observed in NPKM, and following was in order of CK, NPKS, NPK, Fallow. Soil fulvic acid (FA) contents in NPKM, NPKS and Fallow were in range of 2.2 to 2.3 g/kg, significantly higher than NPK and CK. Soil humin (HM) content in NPKM was 18.6 g/kg, significantly higher than other treatments (p<0.05). The order of soil spectral reflectance from high to low was as following NPK > Fallow, CK > NPKS > NPKM. In contrast with CK, soil spectral reflectance in NPK was increased by 6.5% on average, while in NPKS and NPKM were decreased by 11.1% and 15.1% in average, respectively. According to the linear correlation analysis results, the spectral reflectance of black soil was significantly negatively correlated with soil HA and HM contents (p<0.01), and the correlation coefficient (r) were -0.858 and -0.681, respectively. Summing up the results above, it can be concluded that long-term organic materials application could significantly enhance soil humic substance content, lower soil spectral reflectance, and make black soil darker. Oppositely, long-term chemical fertilizer application increased soil spectral reflectance, and made black soil "faded". The effects of organic manure on improving soil organic materials and humic substances were better than straw.