Light microscopical study of endosperm formation in Brassica napus L.

The cellularization of the endosperm of Brassica napus was investigated with light microscopy after embedment in Technovit 7100. The microtubular cytoskeleton was visualized by immunofluorescence techniques after embedment in butyl methyl metacrylate (BMM). The analyses of sectioned seeds, sampled at various developmental stages, revealed that the endosperm has a nuclear phase up to the early heart shaped stage of the embryo. From the heart shaped stage onwards cells and alveoli are formed in the endosperm. The cellularization of endosperm was preceded by alveolus formation except in the region surrounding the embryo suspensor and in the chalazal zone of the embryo sac where the nuclear endosperm vacuolated and cell walls were formed all around the nuclei. Alveolus formation only occurred when nuclei had attained a dense distribution in the layer of cytoplasm aligning the wall of the central cell. When nuclei divided within the alveoli, cell plate formation resulted in the formation of mononuclear endosperm cells along the embryo sac wall. When the walls of the alveoli grew towards the centre of the embryo sac, alveoli regularly closed leaving space for enlargement of the remaining alveoli. In this way endosperm cells enlarged going from the periphery to the central area of the embryo sac. The microtubular cytoskeleton was visualized in the nuclear, alveolar and cellular endosperm. The pattern observed, showed that the organization and function of the microtubular arrays was as generally found during endosperm development.