Two field experiments were conducted to study the effects of 6-year plastic film mulching on bacterial diversity, organic matter of paddy soil and water use efficiency on different soils with great environmental variabilities in Zhejiang Province, China, under non-flooding condition. The experiment started in 2001 at two sites with one rice crop annually. Three treatments included plastic film mulching with no flooding (PM), no plastic film mulching and no flooding (UM), and traditional flooding management (TF). Soil samples were collected and analyzed for bacterial diversity by DGGE and organic matter content, and water use efficiency (WUE) was calculated. The results showed that PM treatment favored the development of a more total bacterial community compared with TF management, the total number of bands was 33.3, 31.7 at tiller stage and heading stage (p < 0.05*). Hence, organic matter content was decreased by 36.7% and 51.4% under PM at two sites. PM also produced similar rice grain yield as TF at Duntou site and Dingqiao site, the average was 7924 kg?ha?1 and 7015 kg?ha?1 for PM and 8150 kg?ha?1 and 6990 kg?ha?1 for TF, respectively. Compared to TF, WUE and irrigation water use efficiency were increased significantly by 70.2% - 80.4% and 273.7% - 1300.0% for PM. It is essential to develop the water-saving agriculture.