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First published online 24 July 2008
doi: 10.1242/dev.023648

Development 135, 2839-2843 (2008)
Published by The Company of Biologists 2008
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Vascular signalling mediated by ZWILLE potentiates WUSCHEL function during shoot meristem stem cell development in the Arabidopsis embryo

Matthew R. Tucker1, Annika Hinze1, Elise J. Tucker1, Shinobu Takada2, Gerd Jürgens2 and Thomas Laux1,*

1 Faculty of Biology, Schaenzlestrasse 1, University of Freiburg, D-79104 Freiburg, Germany.
2 Developmental Genetics, Centre for Molecular Biology of Plants, University of Tübingen, D-72076 Tübingen, Germany.


Figure 1
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  		Fig. 1. pCLV3:GFP-ER expression is altered in zll-1 embryos. (A-C) pCLV3:GFP-ER expression (green) in wild-type (WT) Arabidopsis embryos. (A) Transition stage embryo (B) Close-up of stem cells in a torpedo stage embryo. (C) Bent cotyledon stage embryo. (D-F) pCLV3:GFP-ER expression in zll-1. (D) Transition stage embryo. (E) Close-up of stem cells at torpedo stage showing slightly weaker expression than in wild type. (F) Bent cotyledon stage embryo showing no pCLV3:GFP-ER expression. cp, cotyledon primordium; c, cotyledon; sc, stem cells; L1, L2 and L3, tissue layers 1, 2 and 3, respectively; sam, shoot apical meristem; h, hypocotyl. Scale bars: 10 µm in A,B,D,E; 20 µm in C,F.

 

Figure 2
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  		Fig. 2. pZLL:YFP-ZLL expression relative to the stem cell niche. (A) YFP-ZLL (yellow) accumulates in the apical cell of the 2-cell stage Arabidopsis embryo (blue, DAPI). (B,C) YFP-ZLL localisation relative to WUS-GFP3 (green). (B) 16- to 32-cell stage embryo. (C) Early heart stage embryo. The dashed line indicates the uppermost layer of YFP-ZLL-expressing cells. (D,E) YFP-ZLL localisation relative to pCLV3:CFP-ER expression (cyan). (D) Early heart stage embryo. (E) Late heart stage embryo. The dashed line in E indicates the same boundary as in C. (F) Pro-vascular pATHB8:YFP-ZLL (yellow) expression does not overlap with pCLV3:CFP-ER (pink) or gWUS-GFP3 (white) expression during embryogenesis. A torpedo stage embryo is shown. Insets show the differential contrast/fluorescent overlay. c, cotyledon; sam, shoot apical meristem; vp, vascular primordium; ad, adaxial cotyledon domain. Scale bars: 5 µm.

 

Figure 3
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  		Fig. 3. ZLL functions from outside of the stem cells during embryogenesis. Expression of (A-C) pZLL:YFP-ZLL, (D-F) pCLV3:YFP-ZLL, (G-I) pAS2:YFP-ZLL, (J-L) pATHB8:YFP-ZLL, (M-O) pARR5:YFP-ZLL and (P-R) pAS1:YFP-ZLL in zll-1 mutants. Transition/early heart and torpedo stage Arabidopsis embryos are shown, and seedlings are at 16 days post-germination. c, cotyledon; sam, shoot apical meristem; vp, vascular primordium; l, leaf; sc, stem cells; tm, terminated meristem. Scale bars: 10 µm, except 2.5 mm in C,F,I,L,O,R.

 

Figure 4
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  		Fig. 4. WUS expression and function is altered in zll-1 mutant embryos. (A-C) gWUS-GFP3 expression in wild-type (WT) Arabidopsis embryo. (A) 16-cell stage showing WUS-GFP3 (green). (B) Late heart stage embryo showing WUS-GFP3 marking the organising centre. (C) Walking-stick stage embryo. The meristem dome is indicated with a dashed line. Inset shows endogenous WUS mRNA localisation in an embryo at the same stage. (D-F) gWUS-GFP3 in zll-1. (D) 16-cell stage embryo. (E) Late heart stage embryo. (F) Walking-stick stage embryo showing a broad domain of gWUS-GFP3 expression. The flat meristem dome is indicated with a dashed line. Inset shows endogenous WUS mRNA localisation at the same stage. (G) Wild-type seedling. (H) Wild-type ATRPS5A>>WUS seedling showing ectopic outgrowths on the hypocotyl (arrows) and stunted cotyledons. (I) zll-1 seedling. (J) zll-1 ATRPS5A>>WUS seedling. All seedlings shown are at 11 days post-germination. c, cotyledon; sam, shoot apical meristem; h, hypocotyl; l, leaf; ts, terminal structure. Scale bars: 10 µm in A-F; 0.5 mm in G-J.

 

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