Abstract: Anthers of tobacco varieties Honghuadajinyuan and Yunyan 97 were dried to detect the impact of different drying conditions on pollen viability. Pollen collection rate and viability were determined after dried at the temperature of 10℃, 25℃, 30℃, 40℃, 50℃, for 12 h, 24 h, 36 h, and 48 h. These samples were then stored for 12 h, 24 h, and 36 h at 4℃, and checked for viability every 12 hours. The results showed that, (1) The pollen collection rate at different drying temperature of Honghuadajinyuan and Yunyan 97 went down as the drying time prolonged, both reached the highest at 12 h. (2) Honghuadajinyuan pollen dried under 10℃ and 25℃, and Yunyan 97 pollen dried under 25℃ had higher vigor, while both pollen viability decreased as drying temperature increasing and time prolonging when drying temperature surpassed 25℃ for Honghuadajinyuan and 10℃ for Yunyan 97. (3) The pollen vigor which collected at the same drying temperature decreased as storage time increasing at 4℃, and dropped faster along with drying time. Pollen viability collected at 25℃ declined more slowly than at 10℃, which was more fitful for storage. (4) Honghuadajinyuan and Yunyan 97 pollens collected at 25℃, 12 h could be stored for 36 h at 4℃ with the highest viability. In summary, the most suitable drying condition of Honghuadajinyuan and Yunyan 97 pollen was 25℃ and 12 h, which could guarantee the highest collection rate, pollen viability, and pollen storability.