Soil continuous cropping obstacles lead to yield and economic losses in agriculture. Reductive soil disinfestation (RSD) is an effective technology for alleviating it. However, the key factors influencing microbial community composition and how do they promote functional transformation of core microbes under RSD practice remain poorly understood. Hence, a short-term field experiment was performed integrating real-time polymerase chain reaction, average well color development (AWCD), and MiSeq pyrosequencing to investigate relationships between environmental factors and microorganisms in five different disinfestation treatments, i.e., untreated monoculture soil (CK), soil with high temperature heating (HT), soil with dazomet (DZ), RSD with sugarcane bagasse (SB), or with bean dregs (BD). The results showed that compared to non-RSD treatments, both RSD treatments significantly increased soil microbial abundance and soil available K and organic matter (OM). Further analysis found that available K and OM were the key factors inducing microbial community change. Additionally, relative to non-RSD treatments, the relative abundances of phyla Proteobacteria, Acidobacteria, Rokubacteria, and Ascomycota were higher, whereas those of Actinobacteria, Gemmatimonadetes, and Basidiomycota were lower in RSD treatments. Changes in microbial diversity and abundance led to variation of soil microbial community functions. AWCD and community function prediction showed that, in contrast with non-RSD treatments, soil metabolism activity significantly increased, bacterial community functions including terpenoids and polyketides metabolism, signal transduction and cell motility increased, and the number of saprotroph fungi increased under RSD treatments. Overall, RSD incorporated with sugarcane bagasse or bean dregs efficiently improved soil fertility, and considerably increased soil microbial activity and function, which may benefit future sustainable agriculture production.