Control of plant cell fate transitions by transcriptional and hormonal signals
- University of Heidelberg, Germany
- Heidelberg University, Germany
- University of California, San Diego, United States
Figures
Figure 1
SAM size and organ initiation rate are uncoupled in hec1,2,3.
(A–B) Representative views of 3D-reconstructed shoot meristems after nuclei segmentation from WT (A) and hec1,2,3 (B). Light blue: SAM cells; dark blue: primordia cells. (C) Shoot apical meristem …
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Figure 1—source data 1
SAM size quantification (panel C);
SAM cell number quantification (panel D); Quantification silique number (panel G); Mean plastochron quantification (panel H).
- https://doi.org/10.7554/eLife.30135.004
Figure 2 with 6 supplements
HEC function controls CZ to PZ fate transition.
(A–B) Representative expression of pWUS:3xYFP-NLS in SAMs of pCLV3:HEC1-linker-GR plants four days after mock (A) or dex (B) treatment. (C–D) Development of WUS (C) and CLV3 (D) domain sizes in pCLV3…
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Figure 2—source data 1
Intensity plot profiles: pWUS:3xYFP-NLS (panel C).
Intensity plot profiles: pCLV3:mCherry-NLS (panel D); CLV3 cell number quantification (panel G); Ratio CLV3/SAM cells: pCLV3:HEC1-linker-GR (Figure 2H); Mitotic index CZ quantification (panel K); Mitotic index PZ quantification (panel L).
- https://doi.org/10.7554/eLife.30135.012
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Figure 2—source data 2
SAM size measurement pCLV3:HEC1-linker-GR (Figure 2—figure supplement 2A);
SAM cell number quantification (Figure 2—figure supplement 6B); Mitotic index quantification (Figure 2—figure supplement 6C).
- https://doi.org/10.7554/eLife.30135.013
Figure 2—figure supplement 1
HEC1-linker-GFP protein mobility in the SAM.
(A) Schematic representation of the SAM with CLV3 (yellow), WUS (red) and HEC1 (green) mRNA expression pattern. Central zone (CZ, yellow), organising centre (OC, red), peripheral zone (PZ, light …
Figure 2—figure supplement 2
SAM behavior after promoting of HEC function at the CZ.
(A) Development of individual pCLV3:HEC1-linker-GR SAM size after mock or dex treatment (n > 6 per condition). (B) Representative view of pCLV3:HEC1-linker-GFP SAM 30 DAG. (C–D) Representative views …
Figure 2—figure supplement 3
Dynamics of WUS and CLV3 reporters after HEC1-GR induction at the CZ.
(A) Development of individual pCLV3:HEC1-linker-GR shoot meristems expressing pCLV3:mCherry-NLS and pWUS:3xYFP-NLS after mock or dex induction. pWUS:3xYFP-NLS (green) marks the OC whereas pCLV3:mCher…
Figure 2—figure supplement 4
Intensity plot profiles of WUS and CLV3 reporters after HEC1-GR induction at the CZ.
(A–B) Time series of intensity plot profiles in pCLV3:HEC1-linker-GR plants expressing pCLV3:mCherry-NLS and pWUS:3xYFP-NLS. The plots show the distribution of CLV3 signal (A) and WUS signal (B) in …
Figure 2—figure supplement 5
Time series of 3D-reconstructed pCLV3:HEC1-linker-GR shoot meristems expressing pCLV3:mCherry-NLS/UBQ10:3xGFP-NLS after mock or dex induction.
Yellow, orange and red: CLV3 positive cells; light blue: PZ cells; dark blue: primordia. Note the progressive increase in the SAM and the CZ size after dex treatment. (n > 9 per condition). Scale …
Figure 2—figure supplement 6
Dynamics of the SAM proliferative activity after HEC1-GR induction at the CZ.
(A) Time series of 3D-reconstructed pCLV3:HEC1-linker-GR shoot meristems expressing pKNOLLE:fast-mFluorescentTimer-NLS after mock or dex treatment. Bright red, orange, yellow: young dividing cells; …
Figure 3 with 1 supplement
HEC function controls PZ to organ primordia fate transition.
(A–B) Representative images of pCUC2:HEC1-linker-GR four days after mock (A) or dex treatment (B). (C) Quantification of primordia number formed in pCUC2:HEC1-linker-GR after mock and dex treatment …
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Figure 3—source data 1
Quantification primordia number (panel C).
- https://doi.org/10.7554/eLife.30135.016
Figure 3—figure supplement 1
SAM behaviour after promoting HEC function at the PZ/BZ.
(A) Representative activity of pCUC2:mCherry-GR-linker-NLS in the shoot meristem (n > 3). Green signal marks mCherry-NLS. (B) pCUC2:HEC1-linker-GFP inflorescence 30 DAG (n > 30). Note the pin-like …
Figure 4 with 1 supplement
HEC genes control cell differentiation dynamics in the SAM.
Computational simulations displaying in silico estimation (line) and observed in vivo quantification (boxes) for CZ cell number, total cell number, CZ/total cell ratio, number of unseparated …
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Figure 4—source data 1
Quantification auxin maxima pDR5v2:3xYFP-NLS (panel E).
- https://doi.org/10.7554/eLife.30135.019
Figure 4—figure supplement 1
Computational model simulations.
(A–G) Model simulations displaying in silico estimation (line) and observed in vivo quantification (whisker box) for CZ cell count, total cell count, ratio CZ/total cell count, ratio specified …
Figure 5 with 1 supplement
HEC genes promote cytokinin signalling.
(A–B) Representative expression of pTCSn:erGFP in WT (A) and hec1,2,3 (B) SAMs. (C) Average intensity plot profile of pTCSn:erGFP signal in WT and hec1,2,3 SAM (n > 14 per genotype). (D–F) Analysis …
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Figure 5—source data 1
Intensity plot profiles pTCSn:erGFP (panel C);
TCS quantification plot profile (panel F); Source data also included for panels G and H.
- https://doi.org/10.7554/eLife.30135.022
Figure 5—figure supplement 1
Dynamics of cytokinin responses after modulation of HEC function.
(A–B) Intensity based representation of root apical meristems expressing pTCSn:erGFP in WT (n = 5)(A) and hec1,2,3 (n = 10) (B). (C) Time series of individual pCLV3:HEC1-linker-GR shoot meristems …
Figure 6 with 3 supplements
HEC function modulates auxin signalling.
(A–B) pDR5v2:3xYFP-NLS expression three days after mock (A) or dex treatment (B) of pCUC2:HEC1-linker-GR/pRPS5a:mCherry-NLS plants. (C) Quantification of auxin maxima in pCUC2:HEC1-linker-GR after …
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Figure 6—source data 1
Quantification auxin maxima SAM (panel C).
- https://doi.org/10.7554/eLife.30135.027
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Figure 6—source data 2
Quantification number of auxin maxima (Figure 6—figure supplement 2C);
Root Expression data Li et al., 2016 (Figure 6—figure supplement 3A); RAM size measurement p16:HEC1-linker-GR (Figure 6—figure supplement 3G); Quantification cortex cell number p16:HEC1-linker-GR (Figure 6—figure supplement 3H); Cortex cell lenght measurement p16:HEC1-linker-GR (Figure 6—figure supplement 3J).
- https://doi.org/10.7554/eLife.30135.028
Figure 6—figure supplement 1
Auxin responses at the SAM in HEC loss-of-function mutants.
(A–B) Representative expression of pDR5v2:3xYFP-NLS in WT (n = 20) (A) and hec1,2,3 (n = 17) (B) shoot meristems (intensity based colour coding). White asterisks mark primordia (C–D) Representative …
Figure 6—figure supplement 2
Dynamics of auxin responses at the SAM in HEC gain-of-function mutants.
(A) Time series of pDR5v2:3xYFP-NLS activity in individual pCUC2:HEC1-linker-GR/pRPS5A:dtTomato-NLS shoot meristems after mock or dex treatment (n > 7 per condition).(B) Time series of pDR5v2:3xYFP-N…
Figure 6—figure supplement 3
RAM developmental dynamics in HEC gain-of-function mutants.
(A) Average expression level of HECATE genes and SHORTROOT (SHR) in different root domains as calculated in Li et al. (2016). (B) Representative image of p16:mCherry-GR-linker-NLS root three days …
Figure 7 with 2 supplements
HEC1 DNA binding profile.
(A) Genome-wide distribution of HEC1-bound regions relative to gene models. (B) Motif enrichment analysis from HEC1 ChIP-seq. Position weight matrix depicting G-box, ARE and GARP motifs and …
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Figure 7—source data 1
Calculation ARE_G-box enrichments (Figure 7—figure supplement 1G–I).
- https://doi.org/10.7554/eLife.30135.032
Figure 7—figure supplement 1
Analysis of HEC regulatory patterns in genome-wide profiles.
(A) Early HEC1-response genes as shown by the overlap between genes bound by HEC1 in p35S:HEC1-GFP ChIP-seq and regulated in p16:HEC1-linker-GR RNA-seq experiments. (B) Relative PIN3 expression in p1…
Figure 7—figure supplement 2
Visualization of HEC1 binding peaks at the loci of ARFs and Aux-IAAs expressed in the SAM.
(A) IGV visualization of HEC1 binding peaks at the promoter of ARFs expressed in the SAM. (B) IGV visualization of HEC1 peaks at the promoter of Aux-IAAs expressed in the SAM (Vernoux et al., 2011). …
Figure 8 with 1 supplement
HEC factors interact with MP and unbalance the auxin feedback system.
(A) Yeast-two-Hybrid assay between MONOPTEROS (MP) and HEC1, HEC2 or HEC3. Empty vector: negative control; BODENLOS (BDL): positive control. Blue staining denotes physical interaction. (B) …
Figure 8—figure supplement 1
Dynamics of MP-GFP, PIN1-GFP and R2D2 after HEC1-GR induction at the PZ/BZ.
(A) Time series of pMP:MP-GFP in individual pCUC2:HEC1-linker-GR shoot meristems after mock or dex treatment (n > 6 per condition). (B) Time series of auxin signalling input in individual pCUC2:HEC1-…
Figure 9 with 1 supplement
Auxin feedback stabilization suppresses HEC function at the SAM periphery.
(A–B) Relative expression of MP (A) and SPT (B) 24 hr after p16:HEC1-linker-GR induction (M/M: mock/mock, N/M: NAA/mock, M/D: mock/dex, N/D: NAA/dex) (C–E) Representative views of pCUC2:HEC-linker-GR…
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Figure 9—source data 1
qRT-PCR (panel 9A and B);
Quantification primordia number for pCUC2:HEC1-linker-GR (panel I); Quantification primordia number for p16:HEC1-linker-GR (panel J).
- https://doi.org/10.7554/eLife.30135.037
Figure 9—figure supplement 1
Dynamics of primordia initiation after HEC1-GR induction at the PZ/BZ and auxin co-treatment.
(A–D) Representative view of shoot meristems expressing pCUC2:3xGFP-NLS following 24 hr (A,B) or 48 hr (C,D) of mock (A,C) or NAA (B,D) treatment (n > 3 per condition). (E–F) Development of pCUC2:HEC…
Figure 10 with 1 supplement
HEC function integrates environmental signals at the SAM.
(A–D) Reconstructed views of shoot meristems in wild type (A, C) and hec1,2,3 (B, D) plants grown under normal (A, B) and low light intensity conditions (C, D) (n > 13). (E) Shoot meristem size in …
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Figure 10—source data 1
Source data provided for panel E and intensity plot profiles pTCSn:erGFP under different light regimes (panel J).
- https://doi.org/10.7554/eLife.30135.040
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Figure 10—source data 2
Cell surface area MorphographX (Figure 10—figure supplement 1E);
Cell surface area: MorphoGraphX individual plants (Figure 10—figure supplement 1F).
- https://doi.org/10.7554/eLife.30135.041
Figure 10—figure supplement 1
Cell size at the SAM after transfer to low light conditions.
(A–D) Cell area representation of wild type (A, C) and hec1,2,3 (B, D) plants grown under normal (A, B) and low light intensity conditions (B, D). Heat map colors depict cell size (µm2). (E–F) Cell …
Figure 11
Theoretical model depicting HEC activity controlling cellular fate transition in the SAM by modulating the auxin-cytokinin phytohormonal balance and integrating environmental signals.
https://doi.org/10.7554/eLife.30135.042Additional files
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Supplementary file 1
ChIPseq, RNAseq and QRT-PCR analyses
- https://doi.org/10.7554/eLife.30135.043
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Supplementary file 2
List of oligonucleotides
- https://doi.org/10.7554/eLife.30135.044
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Supplementary file 3
Description of computational model
- https://doi.org/10.7554/eLife.30135.045
