Due to the different species of Epimedium separated by seasonal isolation and geography, developing a method for midterm storage of E. pubescens is necessary to facilitate germplasm conservation, interspecific hybridization and identification of hybrid species. In order to optimize the process of hand-pollination, in this work we have studied the conservation potential of E. pubescens pollen stored for approximately 2 months at -20°C, -70°C and -196°C. The optimal desiccation time of E. pubescens pollen was found to be 210 min. In vitro pollen germination of fresh pollen was 56.0%, and it was progressively reduced after conservation at -20°C, -70°C and -196°C reaching a minimum of 4.2%, 6.0% and 4.4%, respectively after 2 months of storage. The germination capacity of pollen stored at -20°C and -196°C was similar to that observed at -70°C, therefore for reasons of convenience and economy, pollen can be efficiently stored at -20°C. There were no fruit set differences between the field pollinations performed with stored pollen and pollinations performed with fresh pollen. These results indicate that E. pubescens pollen can effectively stored at subzero temperatures for several weeks to months.