BACKGROUND: Lipids in rapeseed is of great significance to human health, and 'Fangyou 777' (No. GPD-2019-510073) has been identified as an excellent cultivar with high oil content. However, the change of lipid profile at different ripening stages remain unclear. Herein, UPLC-MS/MS was utilized for comprehensive lipidomics analysis of 'Fangyou 777' and its parents at four ripening stages. RESULTS: 778 lipids components across 25 subclasses were identified, and triglycerides (TGs), diglycerides (DGs), phosphatidylserines (PSs), phosphatidylinositols (PIs), phosphatidylglycerols (PGs), phosphatidylcholines (PCs), phosphatidylethanolamines (PEs), and free fatty acids (FFAs) were identified as the dominant lipid subclass. Due to heterotic vigor, the total lipids, TGs, FFAs, lysophosphatidylglycerol (LPGs) and PSs contents in 'Fangyou 777' were significantly higher than its parents. The PCA and OPLS-DA results elucidated that lipids in 'Fangyou 777' differed obviously from its parents at S1 (17 April, 2023
28 days before ripening, 28 DBR), S2 (1 May, 2023
14 DBR), and S3 (15 May, 2023
ripening day). TG(18:1_18:3_22:1), TG(18:1_22:1_18:2), TG(16:0_18:1_20:1), TG(16:0_18:1_22:1), TG(20:1_18:2_20:2), TG(18:1_18:1_20:1), and FFA(24:4) were recognized as key differential lipids. The glycerolipid metabolism and unsaturated fatty acid biosynthesis were the differential metabolic pathways at S1 and S3, while glycosylphosphatidylinositol (GPI)-anchor biosynthesis and glycerophospholipid metabolism were the differential metabolic pathways at S2 and S4 (7 days after ripening/physiologically ripened for one week). CONCLUSION: This study provided a comprehensive profile to facilitate the understanding lipids accumulation in 'Fangyou 777' and its parents during ripening stages, and offered a foundation to comprehend lipid metabolism.