National Monitoring Station of Soil Fertility as a research platform was used combined with UV-Visible absorption spectroscopy to explore how the following fertilizer application: (1) CK (non-fertilization); (2)NPK (applied chemical nitrogen, phosphorus and potassium); (3) NPKS (chemical nitrogen, phosphorus and potassium combined with straw); (4) 1.5NPKS (1.5 times of chemical nitrogen, phosphorus and potassium combined with straw); (5) NPKM (chemical nitrogen, phosphorus and potassium combined with manure) to influence the DOM content and structure in 0~20 and 20~40 and 40~60 cm soil layers. The results showed that long term application chemical fertilizers had less effects on soil DOM content, but chemical fertilizers combined with straw and organic manure significantly increased soil DOM content by 39.1 and 12.1 mg·kg-1 in topsoil. Compared to CK treatment and NPK treatment, chemical fertilizers combined with manure and straw could decrease C/N in 0~20 and 20~40 cm soil layers, and improved the supply of nitrogen in soil, but the DOM and C/N had not significantly difference between different fertilizations in 40~60 cm soil layer. In 0~20 and 20~40 cm soil layers, compared to CK treatment and NPK treatment, the treatments of NPKM, NPKS and 1.5 NPKS increased conjugated structure and humus of soil DOM . In 40~60 cm soil layer, the increased by different fertilizations were not obvious. Chemical fertilizers combined with straw or manure can increase ultraviolet spectrum absorption values of DOM (SUVA254 and SUVA260 and SUVA280) and α (355) in 0~20 cm soil layer, and decrease Ultraviolet spectrum absorption ratios of soil DOM (A250/A365). It showed that the fragrance, hydrophobicity, molecular weight and CDOM increased. Meanwhile, it was not obvious in 20~40 and 40~60 cm soil layers. A300/A400 in different soil layers was greater than 3.5, and it showed this soil was rich in Fulvic Acid. Long term chemical fertilizers combined with manure and straw can increase DOM content in 0~20 and 20~40 cm soil layers, decrease C/N, and increase soil fertility. Meanwhile, it also can increase the conjugated structure, humus, fragrance, hydrophobicity and molecular weight of DOM in 0~20 cm soil layer. Long term fertilization has less effects on soil DOM content and structure in 40~60 cm soil layer.