Structural changes during the first divisions of embryos resulting from anther and free microspore culture inBrassica napus

Summary Ultrastructural and cytochemical features of embryo development during anther and free microspore culture inBrassica napus have been followed from the late uninucleate microspore stage through the first embryonic division. On transfer to culture, the microspore cytoplasm possesses a large vacuole, often containing electron opaque aggregates, and a peripheral nucleus. Mitochondria, endoplasmic reticulum and starch-free plastids are distributed throughout the cytoplasm. The conditions of culture induce a number of major changes in the cytoplasmic organisation of the microspore. First, the central vacuole becomes fragmented allowing the nucleus to assume a central position within the cell. Secondly, starch synthesis commences in the plastids which, in turn, are seen to occupy a domain investing the nucleus. Thirdly, the cell develops a thick fibrillar wall, situated immediately adjacent to the intine of the immature pollen wall. Finally, the microspores develop large cytoplasmic aggregates of globular material. The nature of this substance remains unknown, but it remains present until the young embryos have reached the 30 cell stage. The first division of cultured microspores destined to become embryos is generally symmetrical, in contrast to the asymmetric division seen in normal development in vivo. Consideration is given to the differences observed between embryos developing from anthers and free microspores in culture.