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. 2002 Aug;129(4):1807–1819. doi: 10.1104/pp.000810

Figure 1.

Figure 1

Development of alfalfa subsp. varia A2 leaf protoplast-derived cells cultured at different 2,4-D and Fe (Fe-EDTA) concentrations. Leaf protoplast-derived cells of the embryogenic A2 alfalfa genotype cultured in a normal medium containing 100 μm Fe-EDTA at 1 μm 2,4-D (A and B) or 10 μm 2,4-D (C and D) concentration and in a medium with excess (1 mm) Fe-EDTA in the presence of 1 μm 2,4-D (E and F) at 5 d in culture (A, C, and E) and cells from the same cultures after staining by toluidine blue (B, D, and F). Cells of proembryogenic cell colonies express the AGL-15 epitope (Perry et al., 1999) in their nuclei as a characteristic of embryogenic cells (G, immunolabeling; H, nuclear staining by 4,6-diamidino-2-phenylindole; and I, transmitted light). Stages of somatic embryogenesis initiated from the proembryogenic colonies after decreasing the embryogenic (10 μm) 2,4-D concentration to 1 μm in the medium (J, globular; K, heart; and L, torpedo).