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. 2020 Jan 24;71(9):2612–2628. doi: 10.1093/jxb/eraa041

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© The Author(s) 2020. 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/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 7. — Callose deposition precedes WOX2 gene expression during 35S:BBM-induced somatic embryogenesis. (A) Callose staining (green) on the cotyledon tip in 2-day-old 35S:BBM seedlings. Callose is present in PD located in the primary pit fields (arrow) and in the cell plates of newly divided stomatal meristemoids (open arrow). WOX2:NLS-YFP expression was not detected at this stage. (B) Callose staining (purple) along the cotyledon tip and margin in 4-day-old 35S:BBM seedlings. Inset, higher magnification showing callose (arrow). WOX2:NLS-YFP expression was not detected at this stage. (C, D) Five-day-old (C) and 6-day-old (D) 35S:BBM seedlings showing WOX2:NLS-YFP expression (green) and callose staining (purple) in non-overlapping regions. The cotyledon border is marked by a white line. (E, F) Overview of an 8-day-old seedling explant showing WOX2:NLS-YFP expression (green) at the cotyledon margin (E; inset optical section showing gene expression) and tip (F). PD callose (purple) is not detected at this stage in the embryogenic region. (G, H) Callose (purple) and WOX2:NLS-YFP expression (green) colocalize in the same cells as embryogenic protrusions increase in size (G, 10 d old) and differentiate into somatic embryos (H, 12 d old). Scale bars: (A) 30 µm; (D, E inset, F, G) 50 µm; (B, B inset, C) 20 µm; (E, H) 5 mm.