The soil physicochemical properties and soil microbial communities were affected by different fertilizer management. Fertilizer regime were closely relative to the soil texture and nutrient status in a doublecropping paddy field of southern China. However, there was limited information about the influence of different manure nitrogen (N) input on soil microbial communities in a double-cropping rice (Oryza sativa L.) field. Therefore, the short-term different manure N input rate management on soil bacterial and fungal diversity in a double-cropping paddy field of southern China were studied by using Illumina sequencing and quantitative real-time polymerase chain reaction technology in the present paper. The filed experiment were including 100% N of chemical fertilizer (M0), 30% N of organic manure and 70% N of chemical fertilizer (M30), 50% N of organic manure and 50% N of chemical fertilizer (M50), 100% N of organic manure (M100), and without N fertilizer input as control (CK). The results showed that diversity indices of soil microbial communities with application of organic manure and chemical N fertilizer treatments were higher than that of CK treatment. Application of organic manure and chemical N fertilizer management increase soil bacterial abundance of the phylum Actinobacteria, Proteobacteria and Gammaproteobacteria, and soil fungi abundance of the phylum Basidiomycota and Zygomycota were also increased. Compared with CK treatment, the value of Richness, Shannon and McIntosh indices, and taxonomic diversity were increased with M30, M50 and M100 treatments. This finding demonstrated that M30, M50 and M100 treatments modify soil bacterial and fungal diversity. Therefore, the combined application of organic manure and chemical fertilizer N management could significantly increase the abundance of profitable functional bacteria and fungi species in a doublecropping rice field of southern China.