This study investigated the effects of artificial intelligence (AI)-based chatbot and peer feedback mechanisms integrated into the instructional videos (IVs) as a feedback tool on learning performance and intrinsic motivation of pre-service teachers (PTs) in online learning. The participants were 144 PTs from a university in Turkey. A pretest–posttest quasi-experimental design was adopted in this study. Two experimental (EG-1: Immediately elaborated feedback with a chatbot for IVs; EG-2: Delayed peer feedback with comments for IVs) groups and a control group (teaching with IVs) were selected. To collect qualitative data, a survey consisting of open-ended questions was conducted in the experimental groups. The results showed that the learning performance and intrinsic motivation scores of chatbot-based and peer feedback groups were higher than the scores in the traditional learning group. The implications for AI-powered feedback mechanisms and directions for future studies were discussed in this study.