The effects of continuous Eucalyptus cropping on soil properties and microbial characteristics and the specific factors influencing tree species growth remain elusive. In this study, three Eucalyptus stands of three different ages were selected, and soil nutrients, microbial biomass, enzyme activity, microbial community composition, and diversity were quantified for each. The findings indicated a significant decline in soil pH, soil cation exchange, soil organic matter, and available phosphorus content with the plantation age. Simultaneously, there was an observed increase in soil alkaline hydrolyzed nitrogen content. In addition, urease and acid phosphatase activities did not show a significant difference with age. In spite of this, catalase activity exhibited a decline corresponding to the advancement in plantation age. The carbon and nitrogen content of the soil microbial biomass increased with the progression of Eucalyptus planting time. The high-throughput sequencing data demonstrated a reduction in microbial diversity in Eucalyptus soils as the planting age increased. Interestingly, the microbial community structure exhibited minimal alterations, and did not exhibit a predominantly oligotrophic state overall. In conclusion, the study results showed that short-term successive Eucalyptus cropping exerts a significant negative impact on the soil system.