[Objective] This study aimed to investigate the effects of substrate en- zymes activities on nursing of watermelon seedlings. [Method] The composted mushroom residue was mixed with garden soil according to a certain proportion to prepare nursing substrate for watermelon seedlings. During the nursing, the activity variation in alkaline phosphatase, acid phosphatase, neutral phosphatase and urease was investigated. In addition, the correlations between pH value, total nitrogen con- tent, total phosphorus content and organic matter content in substrate and enzyme activity were studied. At different young seedling stages, the rhizospheric substrates with different formulas were sampled for determination of enzymes activities. [Result] The enzyme activity differed significantly among different substrates. The correlation analysis results showed that the higher the organic matter content and total nitrogen content in substrate are, the higher the urease activity is; the phosphatase activity was significantly related to the organic matter content, total nitrogen content and to- tal phosphorus content in substrate; the pH value of substrate was significantly relat- ed to rhizospheric alkaline phosphatase activity; the shoot dry weight was positively related to urease activity; there was a significant correlation between phosphatase activity and root dry weight. [Conclusion] Through determining enzymes activities in the rhizospheric substrate for nursing watermelon seedlings, the optimum substrate M3 was screened out. The activities of rhizospheric urease, alkaline phosphatase, acid phosphatase and neutral phosphatase in the substrate M3 were all higher than those in the substrate fertilized with manure.