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. 2014 Aug 23;65(20):5903–5918. doi: 10.1093/jxb/eru330

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© The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 3. — Structural features of WT and vtc2 tissues during SE. (A, B) Medial longitudinal sections of a explants at D7I (+2,4-D) showing cell division within the cotyledon. EA, embryo axis; Cot, cotyledon. Arrows show areas of callus initiation. Bars, 250 µm. (A) WT embryo with evidence of some cell division in the cotyledon. (B) In vtc2-5, tissues were re-organizing and beginning to develop callus in the cotyledon. (C, D) In situ hybridization of SHOOTMERISTEMLESS in proliferating tissue at D7I in WT (C) and vtc2-5 (D) using an anti-STM probe (sense probe, inset). Bars, 20 µm. (E–I) TEM images of WT versus vtc2 somatic embryos during induction. (E) A typical whole cell from a WT somatic embryo. Note the lack of mitochondria and endomembrane system relative to vtc2. N, nucleus; Nu, nucleolus; M, mitochondria; O, oil body; C, chloroplast; ER, endoplasmic reticulum. Bar, 2 µm. (F) A chloroplast from a WT SE showing multiple starch granules (S). Bar, 500nm. (G) Extensive endomembranous structures and ribosomes (R) are found in the cytoplasm of a vtc2 somatic embryo cell. G, Golgi; V, vesicle; ER, endoplasmic reticulum; M, mitochondria; O, oil body; CW, cell wall; V, vacuole. Bar, 300nm. (H) A chloroplast from a vtc2 somatic embryo. Starch (S) is rare and smaller than in the WT. Thylakoid membranes are indicated by an asterisk. Bar, 500nm. (I) Many mitochondria (M) in the cytoplasm of a vtc2 somatic embryo cell. Bar, 2 µm. (This figure is available in colour at JXB online.)