Two mechanisms regulate directional cell growth in Arabidopsis lateral roots
- University of Oxford, United Kingdom
- University Carlos III of Madrid, Spain
Figures
Figure 1 with 3 supplements
Inhibition of RAB-A5c function caused increased anisotropy of the CMT array in meristematic cells.
(A,B) Maximum intensity projections of confocal stacks of lateral roots co-expressing YFP:NPSN12 (top) and RFP:TUB6 (middle) either in wild-type (A) or in a RPS5a > Dex > RAB-A5c[N125I] line (B) …
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Figure 1—source data 1
Quantification of CMT anisotropy and mean orientation in the absence and presence of RAB-A5c[N125I].
- https://doi.org/10.7554/eLife.47988.008
Figure 1—figure supplement 1
CMTs in the absence and presence of RAB-A5cNI.
(A,B) Confocal stacks of lateral roots co-expressing YFP:NPSN12 and RFP:TUB6 either in wild-type (A) or in a RPS5a > Dex > RAB-A5c[N125I] line (B) three days after seedlings were transferred to agar …
Figure 1—figure supplement 2
CMT rearrangement occurs during early stages of RAB-A5c[N125I] induction.
(A–F) Confocal stacks of seedlings expressing YFP:NPSN12 (red), RFP:TUB6 (green), and the Dex-inducible RAB-A5c[N125I]. Images were acquired 24 hr (A–C) and 44 hr (D–F) after seedlings were …
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Figure 1—figure supplement 2—source data 1
Quantification of CMT anisotropy in epidermal lateral root cells during early stages of RAB-A5c[N125I] induction.
- https://doi.org/10.7554/eLife.47988.006
Figure 1—figure supplement 3
GCP2 and RAB-A5c localisation in lateral roots.
(A,B) Average time projections of maximum intensity projections of confocal stacks of lateral roots expressing mCherry:CESA1 in a wild-type (A) or RPS5a > Dex > RAB-A5c[N125I] background (B). …
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Figure 1—figure supplement 3—source data 1
Quantification of colocalisation between YFP:RAB-A5c and GCP2:GFP at cell edges.
- https://doi.org/10.7554/eLife.47988.012
Figure 2 with 2 supplements
A 3D FE model of a meristematic root cell.
(A) Morphology of the uninflated model from the outside (left) and in section (right) of a cell with isotropic CMFs at all faces (left, 1), isotropic CMFs at transverse anticlinal and the outer …
Figure 2—figure supplement 1
Stress distribution in in silico meristematic cells.
(A,B) Effect of selective increase or reduction of the shear moduli at cell edges (µedges) compared to shear moduli at faces (µfaces) on stress distribution for simulations shown in Figure 2B,C.
Figure 2—figure supplement 2
Effect of varying model parameters.
(A,B) Effect of variation in fibre fraction on cell morphology when shear moduli at cell edges (µedges) were selectively increased or decreased compared to shear moduli at faces (µfaces) for …
Figure 3 with 1 supplement
Phenotypic interactions between microtubule mutants and RAB-A5c[N125I].
(A–H) Confocal stacks of lateral roots expressing RPS5a > Dex > RAB-A5c[N125I] in wild-type (A,C,D,E,G), erh3-3 (B), pom2-4 (F), or clasp-1 (H) backgrounds; 3 days after transfer to 0.1% DMSO (left) …
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Figure 3—source data 1
Quantification of lateral root thickness in microtubule mutants in the absence or prasence of RAB-A5c[N125I].
- https://doi.org/10.7554/eLife.47988.016
Figure 3—figure supplement 1
Synergistic effect of microtubule inhibitors and RAB-A5c[N125I] expression on lateral root morphology.
(A,B) Maximum intensity projections of confocal stacks of lateral root meristematic cells expressing YFP:NPSN12 (green) and RFP:TUB6 (red) in a wild-type (A,C,E) and RPS5a > Dex > RAB-A5c[N125I] (B,D…
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Figure 3—figure supplement 1—source data 1
Quantification of lateral root thickness in response to treatment with oryzalin and taxol in the absence or presence of RAB-A5c[N125I].
- https://doi.org/10.7554/eLife.47988.015
Figure 4 with 5 supplements
YFP:RAB-A5c localisation and CMT organisation.
(A) Maximum intensity projections of 3D confocal stacks from lateral roots expressing YFP:RAB-A5c (left) and RFP:TUB6 (middle). Scale bar 20 µm. (B,C) Maximum intensity projections of 3D confocal …
Figure 4—figure supplement 1
YFP:RAB-A5c and RFP:TUB6 dynamics during lateral root development.
(A) Maximum intensity projections of a 4D confocal stack from a lateral root expressing YFP:RAB-A5c (green) and RFP:TUB6 (red) at consecutive 40 min time intervals. (B) Close-up of the box shown in …
Figure 4—figure supplement 2
YFP:RAB-A5c localisation in microtubule mutants.
(A,B) Plots as those shown in Figure 4 showing relative enrichment of YFP:RAB-A5c intensity at an edge normalised against mean edge intensity in the respective cell (Figure 4I) and mean transverse …
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Figure 4—figure supplement 2—source data 1
Quantification of YFP:RAB-A5c intensity at cell edges in microtubule mutant backgrounds.
- https://doi.org/10.7554/eLife.47988.020
Figure 4—video 1
3D maximum intensity projections of 3D confocal stacks from lateral roots expressing YFP:RAB-A5c (top, green) and RFP:MBD (middle, red).
https://doi.org/10.7554/eLife.47988.021
Figure 4—video 2
3D maximum intensity projections of 3D confocal stacks from lateral roots expressing YFP:RAB-A5c (top, green) and RFP:MBD (middle, red).
https://doi.org/10.7554/eLife.47988.022
Figure 4—video 3
Maximum intensity projections of a 4D confocal stack from a lateral root expressing YFP:RAB-A5c (green) and RFP:TUB6 (red) at consecutive 40 min time intervals.
https://doi.org/10.7554/eLife.47988.023
Figure 5 with 2 supplements
Two mechanisms drive growth anisotropy in meristematic lateral root cells.
(A,B) Snapshots from time-lapse confocal series of two lateral roots expressing YFP:NPSN12 at 0 hr (top) and 6 hr (middle, bottom). Epidermal cells were segmented in 3D using MorphoGraphX (Barbier …
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Figure 5—source data 1
Quantification of directional growth in epidermal cells of lateral roots.
- https://doi.org/10.7554/eLife.47988.027
Figure 5—figure supplement 1
Cell growth rates in lateral roots.
(A) Plot showing relative volume growth for roots shown in Figure 5A along the longitudinal axis of the roots. The dotted line corresponds to 1.5-fold volume growth, which was identified manually as …
Figure 5—figure supplement 2
YFP:RAB-A5c localisation in young primary roots.
(A–C) Maximum intensity projections of confocal stacks from 2d old seedlings expressing YFP:RAB-A5c (left) and mCHerry:NPSN12 (middle). (B) Arrows indicate cell plates. Note that edge label is most …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Arabidopsis thaliana) | RAB-A5c/ARA4 | PMID: 1748311 PMID: 26906735 | AT2G43130 | |
| Gene (Arabidopsis thaliana) | KTN | PMID: 11283338 | AT1G80350 | |
| Gene (Arabidopsis thaliana) | CLASP | PMID: 17873093 | AT2G20190 | |
| Gene (Arabidopsis thaliana) | CSI1/POM2 | PMID: 20616083 | AT2G22125 | |
| Gene (Arabidopsis thaliana) | TUB6 | PMID: 1498609 | AT5G12250 | |
| Gene (Arabidopsis thaliana) | GCP2 | PMID: 17714428 | AT5G05620 | |
| Gene (Arabidopsis thaliana) | CESA1/RSW1 | PMID: 9445479 | AT4G32410 | |
| Strain, strain background (Arabidopsis thaliana) | WT; Wild- Type; Col0 | NASC | Nasc stock number: N1093 | |
| Genetic reagent (Arabidopsis thaliana) | erh3-3 | PMID: 11782406 | ||
| Genetic reagent (Arabidopsis thaliana) | spr3 | PMID: 19509058 | ||
| Genetic reagent (Arabidopsis thaliana) | clasp-1 | PMID: 17873093 | ||
| Genetic reagent (Arabidopsis thaliana) | pom2-4 | PMID: 22294619 | ||
| Genetic reagent (Arabidopsis thaliana) | RPS5a > Dex > RAB-A5c[N125I] | PMID: 26906735 | ||
| Genetic reagent (Arabidopsis thaliana) | RAB-A5c::YFP:RAB-A5c | PMID: 26906735 | ||
| Genetic reagent (Arabidopsis thaliana) | UBQ10:: YFP:NPSN12 | PMID: 19309456 | ||
| Genetic reagent (Arabidopsis thaliana) | UBQ10:: mCherry:NPSN12 | PMID: 19309456 | ||
| Genetic reagent (Arabidopsis thaliana) | CLASP::GFP:CLASP | PMID: 17873093 | ||
| Genetic reagent (Arabidopsis thaliana) | UBQ1::RFP:TUB:6 | PMID: 21847104 | ||
| Genetic reagent (Arabidopsis thaliana) | GCP2::GCP2:3xGFP | PMID: 20935636 | ||
| Genetic reagent (Arabidopsis thaliana) | p35S::RFP:MBD | This paper | ||
| Chemical compound, drug | Oryzalin | Sigma-Aldrich | CAS: 19044-88-3 | |
| Chemical compound, drug | Taxol | Sigma-Aldrich | CAS: 33069-62-4 | |
| Chemical compound, drug | Dexamethasone | Sigma-Aldrich | CAS: 50-02-2 | |
| Software, algorithm | Fiji (Fiji is just ImageJ) | PMID: 22743772 | https://imagej.net/Fiji | |
| Software, algorithm | MorphoGraphX | PMID: 25946108 | https://www.mpipz.mpg.de/MorphoGraphX |
Additional files
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Source code 1
Computational model, abaqus subroutine.
- https://doi.org/10.7554/eLife.47988.028
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Transparent reporting form
- https://doi.org/10.7554/eLife.47988.029
