Fig. 3. Cellular monitoring during in vitro anther culture. Microspore development (a–d) and proembryos formation (e–h). Toluidine blue staining (a, b, d–g), DAPI staining
for DNA under confocal microscope (c), and iodide-staining for starch (h). (a) Vacuolate microspore at the beginning of the culture; (b and c) microspore-derived two-cell
structures after one week of culture; (d) multicellular pollen grain with dense cytoplasms after two weeks of culture; (e) panoramic view of an anther after 2 months in
culture showing a rounded multicellular mass or proembryo (arrow); (f) proembryo surrounded by callus cells; (g) detail of proembryo cells; (h) starch cytochemistry, the
proembryo cells (in the centre) show small and scarce starch granules whereas the surrounding cells exhibit larger and more abundant starch inclusions. Bars represent in
(a–d): 10 m, in (e, f and h): 100 m, in (g): 50 m.
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