Leaf Oil Body Functions as a Subcellular Factory for the Production of a Phytoalexin in Arabidopsis

Oil bodies are intracellular structures present in the seed and leaf cells of many land plants. Seed oil bodies are known to function as storage compartments for lipids. However, the physiological function of leaf oil bodies is unknown. Here, we show that leaf oil bodies function as subcellular factories for the production of a stable phytoalexin in response to fungal infection and senescence. Proteomic analysis of oil bodies prepared from Arabidopsis (Arabidopsis thaliana) leaves identified caleosin (CLO3) and a-dioxygenase (a-DOX1). Both CLO3 and a-DOX1 were localized on the surface of oil bodies. Infection with the pathogenic fungus Colletotrichum higginsianum promoted the formation of CLO3-and a-DOX1-positive oil bodies in perilesional areas surrounding the site of infection. a-DOX1 catalyzes the reaction from a-linolenic acid (a major fatty acid component of oil bodies) to an unstable compound, 2-hydroperoxyoctadecatrienoic acid (2-HPOT). Intriguingly, a combination of a-DOX1 and CLO3 produced a stable compound, 2-hydroxyoctadecatrienoic acid (2-HOT), from a-linolenic acid. This suggests that the colocalization of a-DOX1 and CLO3 on oil bodies might prevent the degradation of unstable 2-HPOT by efficiently converting 2-HPOT into the stable compound 2-HOT. We found that 2-HOT had antifungal activity against members of the genus Colletotrichum and that infection with C. higginsianum induced 2-HOT production. These results defined 2-HOT as an Arabidopsis phytoalexin. This study provides, to our knowledge, the first evidence that leaf oil bodies produce a phytoalexin under a pathological condition, which suggests a new mechanism of plant defense.