DEV023648 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 - Adobe PDF

• Supplemental Table S2 - Adobe PDF

• Supplemental Table S3 - Adobe PDF

• Supplemental Table S4 - Adobe PDF

• Supplemental Table S5 - Adobe PDF

• Supplemental Table S6 - Adobe PDF

• Supplemental Table S7 - Adobe PDF

• Supplemental Table S8 - Adobe PDF

• Supplemental Figure S1 -

  Fig. S1. pZLL:YFP-ZLL is expressed in the vascular primordium, adaxial side of the cotyledons and the shoot meristem during late embryogenesis. (A-A′′) Mature zll-1 embryo expressing pZLL:YFP-ZLL (green). The shoot apical meristem (SAM) is outlined with a dotted line. c, cotyledon; h, hypocotyl; vp, vascular primordium.

• Supplemental Figure S2 -

  Fig. S2. CLV3 expression is not induced by ectopic expression of ZLL from the AGO1 promoter. (A,A′) pAGO1:ZLL-YFP expression in transition and torpedo stages. (B) Section of a WT late torpedo embryo showing hybridisation of CLV3 antisense probe in the stem cell region. (C) WT late torpedo embryo showing pCLV3:GFP-ER expression. (D) Section of a zll-1; pAGO1:ZLL late torpedo embryo showing hybridisation of CLV3 antisense probe in the stem cell region, similar to WT. (E) zll-1; pAGO1:ZLL late torpedo embryo showing similar pCLV3:GFP-ER expression to WT. c, cotyledon; SAM, shoot apical meristem; sc, stem cells; AS, antisense.

• Supplemental Figure S3 -

  Fig. S3. ATRPS5A>>WUS mRNA is induced to similar levels in WT and zll-1 embryos. (A-B′) ATRPS5A promoter activity, measured by dsRED2 expression (ATRPS5A>>dsRED2), is indistinguishable between WT and zll-1 embryos. (C) Semi-quantitative RT-PCR of WUS mRNA accumulation in WT; ATRPS5A>>WUS and zll-1; ATRPS5A>>WUS torpedo stage embryos. Thirty cycles of PCR were used to detect WUS and 26 cycles were used to detect the ACTIN 7 (ACT) control. The WUS primers amplify a larger product from WT genomic DNA (gDNA), and this control was used to verify that the RNA contained no detectable endogenous DNA. Endogenous levels of WUS transcript were only detectable after 38 cycles of PCR (*).

• Supplemental Figure S4 -

  Fig. S4. WUS and CLV3 expression are altered during embryogenesis in ago1-1 mutants. (A-C) gWUS-GFP₃ in ago1-1, with insets showing WT siblings. (A) Late heart stage embryo. (B) Walking-stick stage embryo, enlarged to show the meristematic region. (C) Mature ago1-1 embryo. (D) WT sibling of C. (E-G) pCLV3:GFP-ER in ago1-1, with insets showing WT siblings. (E) Heart stage embryo showing no detectable CLV3 expression. (F) Walking-stick stage embryo, enlarged to show the meristematic region. (G) Mature ago1-1 embryo. (H) WT sibling of G. sam, shoot apical meristem. Scale bars: 10 µm in A,B,E,F; 50 µm in C,D,G,H.