In order to investigate the response characteristics of the soil fungal community to short-term fertilization in Eucalyptus artificial forest, Illumina high-throughput sequencing and FUNGuild platforms were performed in this study. The soil fungal community composition, functional groups and their impact factors were analyzed under the short-term fertilization and non-fertilization (control, CK) treatments. As the result shown, Basidiomycota and Ascomycota were identified as the dominant fungi at the class level, accounting for 48.61%-46.74% and 39.31%-38.08%, respectively. The soil fungal community at the class level was not significantly changed by short-term fertilization. On the order level, the relative abundance of (23.73%) and Eurotiales (19.95%) was significantly increased by 7.65 and 1.72 times under the fertilization treatment, respectively, when compared with CK treatment. However, the ratios of Archaeorhizomycetales, Onygenales, Tremellales, and other dominant orders were obviously decreased in soil under the fertilization treatment. The alpha diversity and abundance index of soil fungi were slightly decreased under the fertilization treatment. Meanwhile, the principal coordinate analysis indicated that the fungal community composition was significant difference between the fertilization and CK treatments. Spatrotroph group was identified as the most dominate trophic group in the soil of Eucalyptus artificial forest. The ratio of symbiotroph group was significantly increased by 4.19 times under the fertilization treatment when compared with CK treatment. In addition, the ectomycorrhizal fungus Pisolithus was mainly increased in the symbiotroph group, whereas the relative abundance of other trophic group fungi was obviously decreased. Moreover, the redundancy analysis showed that the soil fungal community was mainly influenced by the soil factors including the contents of soil available nutrition, soil organic carbon, microbial biomass carbon, pH value, and soil moisture. Our results indicated that the short-term fertilization treatment altered the soil fungal community structure and functional group in Eucalyptus artificial forest, and also increased the proportion of symbiotic fungi and improved the soil quality.