An ancient irrigated paddy soil from the Neolithic age was excavated at Chuodunshan Site in the Yangtze River Delta, close to Suzhou, China. The soil organic matter (SOM) content in the prehistoric rice soil is comparable to the average SOM content of present rice soils in this region, but it is about 5 times higher than that in the parent materials. As possible biomarkers to indicate the presence of the prehistoric paddy soil, the bacterial communities were investigated using the techniques of aerobic and anaerobic oligotrophic bacteria enumeration, Biolog analysis, and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The results showed that in the buried soil layers, the prehistoric paddy soil had the largest number of aerobic and anaerobic oligotrophic bacteria, up to 6.12 and 5.86 log cfu g-1 dry soil, respectively. The prehistoric paddy soil displayed better carbon utilization potential and higher functional diversity compared to the parent materials and a prehistoric loess layer. The Shannon index and richness based on DGGE profiles of bacterial 16S rRNA genes were higher in prehistoric paddy soil than those in the prehistoric loess soil. It might be concluded that the prehistoric irrigated rice cultivation accumulated the SOM in plowed soil layer, and thus increased soil bacterial populations, metabolic activity, functional diversity and genetic diversity. Bacterial communities might be considered as the sensitive indicators of the presence of the prehistoric paddy soil in China’s Yangtze River Delta. An ancient irrigated paddy soil from the Neolithic age was excavated at Chuodunshan Site in the Yangtze River Delta, close to Suzhou, China. The soil organic matter (SOM) content in the prehistoric rice soil is comparable to the average SOM content of present rice soils in this region, but it is about 5 times higher than that in the parent materials. As possible biomarkers to indicate the presence of the prehistoric paddy soil, the bacterial communities were investigated using the techniques of aerobic and anaerobic oligotrophic bacteria enumeration, Biolog analysis, and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The results showed that in the buried soil layers, the prehistoric paddy soil had the largest number of aerobic and anaerobic oligotrophic bacteria, up to 6.12 and 5.86 log cfu g-1 dry soil, respectively. The prehistoric paddy soil displayed better carbon utilization potential and higher functional diversity compared to the parent materials a nd a prehistoric loess layer. The Shannon index and richness based on DGGE profiles of bacterial 16S rRNA genes were higher in prehistoric paddy soil than those in the prehistoric loess soil. It might be concluded that the prehistoric irrigated rice cultivated the SOM in plowed soil layer, and increased soil bacterial populations, metabolic activity, functional diversity and genetic diversity. Bacterial communities might be considered as the sensitive indicators of the presence of the prehistoric paddy soil in China’s Yangtze River Delta.