We have conducted the study of ultrastructural changes of wall and cytoplasm of Plantago lanceolata (English plantain) pollen grains during the first 10 min of hydration and activation processes, prior to germination, and localisation of Pla l 1, the major allergen of these pollen grains with immunocytochemical methods. The samples were fixed using conventional and freezing protocols for transmission electron microscopy. During the activation process, the intine is thickened in the apertural region and cytoplasm undergoes changes in the number of lipid bodies, amyloplasts, vacuoles and ribosomes. Also, we observed an association between lipid bodies, cisternae of rough endoplasmic reticulum, dictyosomes and vacuoles. An increase in the presence of allergenic particles (Pla l 1) in the exine, intine and the cytoplasm in activated pollen grains was detected, whereas this presence is not significant in mature pollen grains. The increase in the production and release of this allergen when pollen grains are activated suggest that Pla l 1 has an important role in pollen–stigma recognition and in the subsequent development of the pollen tube.