Abstract: The subtropical zone in China is one of the regions most affected by nitrogen deposition. Soil dissolved organic matter (DOM) is considered to be an important indicator of soil organic matter. Nitrogen deposition may alter the quality and quantity of soil DOM by changing soil microbial activity. In this study, we explored the effects of nitrogen addition on soil DOM content, its spectral characteristics and microbial extraceller enzyme activity in the Moso bamboo plantations by setting control (CT), low-nitrogen (LN), and high-nitrogen (HN) addition levels for three-year nitrogen addition. The results showed that there was no significant change in soil pH, dissolved organic carbon, dissolved organic nitrogen, and aroma index following nitrogen addition, while the humification index increased significantly, microbial enzyme activities increased first and then decreased with the increases of nitrogen addition. Fourier transform infrared spectroscopy results showed that soil DOM had similar absorption peaks in seven regions, and that the absorption peaks of 1000 to 1260 cm^-1 were the strongest, indicating an enhanced amount of polysaccharides, alcohols, carboxyl acids, and esters after nitrogen addition. The results of three-dimensional fluorescence spectroscopy showed that soil DOM structure significantly changed following nitrogen addition, with a decrease in low-molecular substances such as protein-like substances and microbial metabolites and a significant increase in high-molecular substances such as humus-like substances. In general, nitrogen addition made soil nitrogen compatible with microbial requirements. Microorganisms decompose substances that were easily degraded in DOM. The structure of soil DOM was more complex after nitrogen addition. Therefore, short-term nitrogen deposition might be conducive to preserving soil fertility.