Roles for polarity and nuclear determinants in specifying daughter cell fates after an asymmetric cell division in the maize leaf  Kimberly Gallagher, Laurie G. Smith  Current Biology  Volume 10, Issue 19, Pages 1229-1232 (October 2000) DOI: 10.1016/S0960-9822(00)00730-2

Fig. 1 Formation of stomata in maize. The first step in formation of a stomate is an asymmetric division that forms a guard mother cell (GMC). Nuclei in the flanking subsidiary mother cells (SMC) then migrate to become aligned with the GMC. SMCs then divide asymmetrically to form small, lens-shaped subsidiary cells. Following subsidiary cell formation, the GMC divides to produce two guard cells. Current Biology 2000 10, 1229-1232DOI: (10.1016/S0960-9822(00)00730-2)

Fig. 2 Differentiation of subsidiary cells in wild type and four SMC division mutants. (a–d) Markers of subsidiary cell fate in wild-type stomata. (a) Subsidiary cell walls stain pink with toluidine blue O (TBO). (b) Subsidiary cell walls lack UV autofluorescence. (c) Subsidiary cells fail to accumulate anthocyanins in pigmented leaves. (d) Subsidiary cells are differentially stained with sodium cobalt nitrate. (e–g) TBO-stained mutant leaves with white arrowheads indicating daughters of abnormal SMC divisions that differentiated as subsidiaries and black arrowheads indicating those that did not. (e) dcd1; (f) brk1; (g) pan1. (h–o) UV autofluorescence of abnormally divided SMCs with detached daughters (arrowheads) in dcd1;dcd2 double mutants. Guard cells are indicated by arrowheads labelled ‘G’. The sister of each detached daughter is outlined in green; nuclei are false colored in red. In (h,j,l,n) the detached daughter differentiated as a subsidiary and the sister did not; in (I,k,m,o) the sister differentiated as a subsidiary and the detached daughter did not. The scale barsrepresent 100?m. Current Biology 2000 10, 1229-1232DOI: (10.1016/S0960-9822(00)00730-2)

Fig. 3 Intracellular organization of SMCs in wild-type and SMC division mutants. (a) SMCs were examined to determine the proportion having a nucleus aligned with the GMC and the proportion with an actin patch. (b) Schematic summary of intracellular organization in SMCs of each genotype, showing an example of a prophase SMC on the left and an SMC undergoing cytokinesis on the right. Nuclei are shown in yellow, microtubules in green, and actin filaments in red. (c) SMCs undergoing cytokinesis were examined to determine the proportion containing an abnormally oriented phragmoplast in which neither daughter nucleus was aligned with the GMC. TBO stained mature leaves were examined to determine the proportion of abnormal SMC divisions resulting in differentiation of neither daughter as a subsidiary. In (a,c), error bars show standard errors. For further information about this analysis, see Supplementary material. Current Biology 2000 10, 1229-1232DOI: (10.1016/S0960-9822(00)00730-2)

Fig. 4 Model for roles of cell polarity and nuclear determinants in specification of subsidiary cell fate. Brk1 and Pan1 are required for polarization of SMCs; Dcd1 and Dcd2 are required later for phragmoplast guidance. We propose that during prophase, subsidiary cell fate determinants (shown in pink) are a localized to the actin patch along with the nucleus. Following nuclear division, these determinants enter the inner daughter nucleus, and the cell that inherits this nucleus differentiates as a subsidiary cell regardless of its size, shape or position. Current Biology 2000 10, 1229-1232DOI: (10.1016/S0960-9822(00)00730-2)