Abstract: In this paper, the zygotic embryos were used as storage materials to further analyze the changes of physiological and biochemical indexes of the zygotic embryos before and after cryopreservation, and the effects of freezing method and dewatering time on physiological and biochemical indexes were clarified, which provided physiological data support for the establishment of long-term storage methods of fine genetic materials of Fraxinus mandshurica. Under the condition of dehydration for 120 min, the activity of dehydrogenase in zygote embryos was the highest, and the activity of catalase, peroxidase and mass fraction of proline reached the maximum. They were 1 168.85 U·g^-1·min^-1, 338.33 U·g^-1·min^-1 and 394.99 μg·g^-1, respectively. Dehydrogenase activity, proline content, peroxidase activity and catalase activity in zygotic embryo cells were all the minimum values after vitrification. The soluble protein fraction and superoxide dismutase activity in zygote embryos reached the maximum values after storage by slow freezing method, which were 18.82 mg·g^-1 and 361.97 U·g^-1, respectively, which were significantly higher than those by other freezing methods. The activity of zygotic embryos reached the highest value under the conditions of drying and dewatering for 120 min and fast freezing.The results showed that the survival rate and physiological indexes of the zygotic embryos of F. mandshurica were affected by different freezing methods, dehydration time and their interaction. In addition to the decrease of malondialdehyde content, the dehydrogenase activity, antioxidant enzyme activity, proline content and soluble protein content in the zygotic embryo cells of F. mandshurica increased with the increase of dehydration time after being stored in different freezing methods. The survival rates of zygotic embryos and somatic embryos after cryopreservation were 62.26% and 51.68%. Therefore, the method of drying and dewatering for 120 min combined with fast freezing is suitable for cryopreservation of F. mandshurica embryos.