*2.3. Development of Bulblets from Somatic Embryos*

For the development of bulblets, single somatic embryos (SEs), clusters with five somatic embryos each, and a single embryo with cotyledonary leaf obtained from cultures under CW5000K or WW2700K lights were further transferred to fresh MSBM medium in glass bottles as described in Section 2.2. Each bottle contained seven explants. These bottles were incubated in a culture room with a temperature set at 25 ± 2 ◦C, and under light (white fluorescent tubes, illumination intensity of 34 µmol m−<sup>2</sup> s −1 ) with a 16 h light and 8 h dark cycle. Observations of bulblet development in all three types of SEs were recorded after three months of incubation.

Microphotographs of cultures were taken by a digital camera (Nikon D90, Tokyo, Japan). For SEM, samples of bulblets were frozen in liquid nitrogen and observed using a scanning electron microscope (JEOL JSM-6330F, Tokyo, Japan).
