Revisiting the Algal Chloroplast Lipid Droplet: The Absence of an Entity That Is Unlikely to Exist

The precise localization of the lipid droplets and the metabolic pathways associated with oil production are crucial to the engineering of microalgae for biofuel production. Several studies have reported detecting lipid droplets within the chloroplast of the microalga Chlamydomonas reinhardtii, which accumulates considerable amounts of triacylglycerol and starch within the cell under nitrogen deprivation or high-light stress conditions. Starch undoubtedly accumulates within the chloroplast, but there have been debates on the localization of the lipid droplets, which are cytosolic organelles in other organisms. Although it is impossible to prove an absence, we tried to repeat experiments that previously indicated the presence of lipid droplets in chloroplasts. Here, we present microscopic results showing no evidence for the presence of lipid droplets within the chloroplast stroma, even though some lipid droplets existed in close association with the chloroplast or were largely engulfed by the chloroplasts. These lipid droplets are cytosolic structures, distinct from the plastoglobules present in the chloroplast stroma. These results not only contrast with the old ideas but also point out that what were previously thought to be chloroplast lipid droplets are likely to be embedded within chloroplast invaginations in association with the outer envelope of the chloroplast without intervention of the endoplasmic reticulum. These findings point to the intriguing possibility of a tight metabolic flow from the chloroplast to the lipid droplet through a close association rather than direct contact of both organelles.