Quantitative Assessment of the Chloroplast Lipidome

In plants and algae, photosynthetic membranes have a unique lipid composition. They differ from all other cellular membranes by their very low amount of phospholipids, besides some phosphatidylglycerol (PG), and high proportion of glycolipids. These glycolipids are the uncharged galactolipids, that is, mono- and digalactosyldiacylglycerol (MGDG and DGDG), and an anionic sulfolipid, that is, sulfoquinovosyldiacylglycerol (SQDG). In all photosynthetic membranes analyzed to date, from cyanobacteria to algae, protists, and plants, the lipid quartet constituted by MGDG, DGDG, SQDG, and PG has been highly conserved, but the composition in fatty acids of these lipids can vary a lot from an organism to another. To better understand the chloroplast biogenesis, it is therefore essential to know their lipid content. Establishing chloroplast lipidome requires first to purify chloroplast from plant or algae tissue. Here we describe the methods to extract the lipid, quantify the lipid amount of the chloroplast, and qualify and quantify the different lipid classes that might be present in these fractions.