The major environmental factors limiting rice growth and production are osmotic stresses such as drought and high salinity. High osmotic stresses directly disrupt cellular activities, leading to plant growth retardation or death. Plants have various response mechanisms to survive under such stresses. Understanding rice’s stress response mechanisms is necessary to enhance the osmotic stress tolerance of rice. However, assessing specific physiological responses to osmotic stresses is difficult because multiple environmental factors affect rice growth. Here, we describe a simple method for analyzing the osmotic stress responses of rice plants using a hydroponic culture system. This method allows comprehensive gene expression and phenotypic analyses under osmotic stress conditions in rice. Various osmotic stress conditions and samples can be tested simultaneously because this method is small-scale. In addition, the procedure is easy, and highly reproductive results can be obtained.