Pollen germination and subsequent pollen tube elongation are essential for successful land plant reproduction. These processes are achieved through well-documented activation of membrane trafficking and cell metabolism. Despite this, our knowledge of the dynamics of cellular phospholipids remains scarce. In this project, we analyzed the turnover of the glycerolipid composition during the establishment of cell polarity and elongation processes in tobacco pollen and described the proteo-lipid composition of pollen plasma membrane-enriched. To achieve this, we have combined several techniques, such as lipidomics, live-cell microscopy, and plasma membrane isolation coupled with mass spectrometry-based proteomic characterization.