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. 2018 Nov 21;70(3):897–908. doi: 10.1093/jxb/ery413

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© The Author(s) 2018. 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. 1. — Treatment with aminoethoxyvinylglycine (AVG) results in the formation of single guard cells (SGCs). (A) Abaxial epidermis of cotyledons in the wild-type (WT) with or without AVG treatment. Arrows indicate SGCs. Scale bars are 20 μm. (B) E1728 expression in the abaxial epidermis of cotyledons in the wild-type before and after AVG treatment. Arrow indicates SGCs. Scale bars are 100 μm. (C) Abaxial epidermis of a cotyledon in the wild-type with AVG treatment. The confocal z-projection was visualized using propidium iodide staining with a confocal laser-scanning microscope. The yellow arrow indicates a SGC without a central cell wall. White arrows indicate normal stomata with two GCs and a thickened central cell wall. The scale bar is 50 μm. (D) GCs and SGCs with DAPI fluorescence. Scale bars are 5 μm. (E) Quantitative analysis of DAPI fluorescence intensity in GCs and SGCs. The data are means (±SD) (n=10) and the significant difference between SGCs and GCs was determined using Student’s t-test: *P<0.01. (F) Frequency of cotyledons with different numbers of SGCs on the abaxial epidermis of AVG-treated wild-type plants. A total of 50 cotyledons from 25 seedlings were examined. (G) Mean number of SGCs per cotyledon in the wild-type with or without (Mock) AVG treatment. The significant difference between the means was determined using Student’s t-test: *P<0.01. A total of 50 cotyledons from 25 seedlings were examined.