Pollen germination and subsequent pollen tube elongation are cellular processes exhibiting massive activation of membrane trafficking and cell metabolism. However, our knowledge of the dynamics of cellular phospholipids in these processes remains scarce. In this paper, we studied the turnover of the glycerolipid composition during the establishment of cell polarity and elongation processes in tobacco pollen. Moreover, we analyzed the lipid composition of pollen plasma membrane-enriched fraction for the first time.