Understanding of seed germination requirements and storage methods is very important to successfully conserve and restore aquatic vegetation. The main question addressed by the research was germination requirements and suitable seed storage methods of Hydrocharis dubia seeds. Furthermore, the water content and respiration rate of H. dubia seeds were studied under different storage conditions. The study found that light and high seed clustering density had a positive effect on germination, while burial had a negative effect. Germination percentages were 60.67 ± 6.11% and 28.40 ± 6.79% in light and dark conditions, respectively. Under clustering densities of 1 and 50, germination percentages were 6.00 ± 2.00% and 59.33 ± 0.67%, respectively. Germination percentages were 50.40 ± 5.00%, 3.20 ± 3.20%, and 0.80 ± 0.80% at depths of 0, 2, and 3 cm, respectively. Oxygen, water level, and substratum had no significant effect on seed germination. Storage method had a significant effect on seed germination, moisture content, and respiration rate. The germination percentages were 64.00 ± 1.67%, 85.20 ± 5.04%, and 92.80 ± 4.27% under the storage conditions of 4 °C-Dry, 4 °C-Wet, and Ambient water temperature-Wet for 2 years, respectively. The seeds had no germination under the storage conditions of Ambient air temperature-Wet and Ambient air temperature-Dry. Overall, the study indicates that seed germination of H. dubia is restricted by light, burial depth, and seed clustering density. Additionally, it was found that H. dubia seeds can be stored in wet environmental conditions at ambient water temperature, similar to seed banks. Specifically, the seeds can be stored in sand and submerged underwater at ambient water temperatures ranging from 4 °C to 25 °C. This study will help with the conservation and restoration of aquatic plants, such as H. dubia.