Plant extracellular vesicles (EVs) are membrane-enclosed nanoparticles that play diverse roles in plant development and response. Recently, impressive progress has been made in the isolation and identification of the proteins and RNAs carried in plant EVs; however, the analysis of EV lipid compositions remains rudimentary. Here, we performed lipidomic analysis of Arabidopsis rosette leaf EVs, revealing a high abundance of certain groups of lipids, in particular sphingolipids, in the EVs. Remarkably, the EV sphingolipids are composed of nearly pure glycosylinositolphosphoceramides (GIPCs), which are green lineage abundant and negatively charged. We further showed that the Arabidopsis TETRASPANIN 8 (TET8) knockout mutant has a lower amount of cellular GIPCs and secrets fewer EVs, companied with impaired reactive oxygen species (ROS) burst toward stresses. Exogenous application of GIPCs promoted the secretion of EVs and ROS burst in both the WT and tet8 mutant. The characteristic enrichment of sphingolipid GIPCs provides valuable insights into the biogenesis and function of plant EVs.

植物细胞外囊泡（EVs）是膜包裹的纳米颗粒，在植物发育和响应中起着多种作用。最近，在分离和鉴定植物电动汽车中携带的蛋白质和RNA方面取得了令人瞩目的进展。然而，对EV脂质组成的分析仍然是基本的。在这里，我们对拟南芥玫瑰叶片电动车进行了脂质组学分析，揭示了电动汽车中某些类脂特别是鞘脂的丰度很高。值得注意的是，EV鞘脂由几乎纯净的糖基肌醇磷酸神经酰胺（GIPC）组成，这些谱系富含绿色谱系并且带负电。我们进一步证明了拟南芥TETRASPANIN 8（TET8）敲除突变体的细胞GIPC数量较少，分泌的EV较少，同时伴随着朝向压力爆发的活性氧（ROS）受损。GIPCs的外源应用促进了WT和tet8突变体中EV的分泌和ROS爆发。鞘脂GIPCs的特征富集为植物EV的生物发生和功能提供了有价值的见解。