Raising rice seedlings in flat trays has become the main method for mechanized transplanting of rice in China. However, seedling blocks raised by this method were easily cracked in practice, and this problem can be solved by padding a thin ramie fiber nonwoven film on the bottom surface of seedling tray. This study was conducted to determine the effects of this film on root-zone environment of rice seedlings. The results showed that on the 10th day after sowing, the soil inorganic nitrogen, especially nitrate nitrogen, content in the root-zone of the film treatment were considerably higher than in the no-film treatment, in contrast, the soil organic matter content was lower in the film treatment, and by the 20th day, the gap between treatments was enlarged. After applying the film, the Chao 1 index and Shannon index values for the soil bacterial community diversity decreased, and the rice seedlings were shorter, had higher root/shoot ratios, lower nitrate contents, and higher soluble sugar contents. We conclude that application of the ramie fiber nonwoven film resulted in substantial changes in the soil nutrient and bacterial community in root-zone in a short time, which significantly impacted the growth and development of rice seedlings.