Abstract: In order to screen the key factors affecting the cross-pollination of Aeonium Series succulents and optimize the artificial cross-pollination technology. In this paper, the pollen and stigma activity of seven Aeonium Series succulent varieties were detected by TTC staining, in vitro germination and benzidine-hydrogen peroxide method to screen the appropriate pollination time and cross parent; 'Voodoo' (♀) × 'Green Velour' (♂) as a cross-pollination combination, the effects of different pollination methods, gibberellin concentration, wet bag interval on the changes of pistil oxidase activity and malondialdehyde content were compared to explore the key techniques for artificial cross-pollination of Aeonium Series succulent plants. The results showed that the pollen and stigma activity was strongest in most varieties at the early flowering stage, which is a suitable period for pollination. Pollen and stigma activity varied significantly among varieties, with 'Voodoo', 'Green Velour' and 'Short Black' having higher pollen vigor and 'Black Kingkong' and 'Ink Painting' were lower. The stigmas were more fertile for 'Voodoo', 'Short Black' and 'Black Kingkong', and 'Green Velour', 'Halloween' were less fertile. Gibberellin concentration, wet bag interval and pollination method had significant effects on pistil oxidase activity and malondialdehyde content after hybridization. The results of ANOVA showed that 30 mg/LGA, 2 h wet bag interval and three treatments of repeated pollination were the most effective, and POD, SOD, β-1,3-glucanase activities were at high levels in pistils and MDA content was lower in 4～24 h after cross-pollination. The results of the study provide a reference for further exploring the physiological and biochemical mechanisms of succulent plants after hybridization, which can help to optimize the cross-pollination technology of succulent plants and improve the hybridization efficiency.