Microalgae have outstanding abilities to transform carbon dioxide (CO2) into useful lipids, which makes them extremely promising as renewable sources for manufacturing beneficial compounds. However, during this process, reactive oxygen species (ROS) can be inevitably formed via electron transfers in basal metabolisms. While the excessive accumulation of ROS can have negative effects, it has been supported that proper accumulation of ROS is essential to these organisms. Recent studies have shown that ROS increases are closely related to total lipid in microalgae under stress conditions. However, the exact mechanism behind this phenomenon remains largely unknown. Therefore, this paper aims to introduce the production and elimination of ROS in microalgae. The roles of ROS in three different signaling pathways for lipid biosynthesis are then reviewed: receptor proteins and phosphatases, as well as redox-sensitive transcription factors. Moreover, the strategies and applications of ROS-induced lipid biosynthesis in microalgae are summarized. Finally, future perspectives in this emerging field are also mentioned, appealing to more researchers to further explore the relative mechanisms. This may contribute to improving lipid accumulation in microalgae.