A long-term experiment was set up in Yingtan of Jiangxi Province to investigate the effects of long-term application of inorganic fertilizers on the biochemical properties of a rice-planting soil derived from Quaternary red earth. Noncultivated soils are extremely eroded and characterized by a low pH and deficiencies in available nutrients, in particular P and N. After 13 years of inorganic fertilization in cultivation for double-cropped rice, the biochemical properties of the soil were changed. The nitrification potential and urease activity were higher in the treatments with N application than those without N application. Acid phosphatase activity and dehydrogenase activity were also higher in the treatments with P application than in those without P application. The dehydrogenase activity correlated well with the concentrations of both total P and hydrolysable N and with rice crop yield, suggesting that dehydrogenase activity might be a suitable indicator for improvement in soil fertility.