CASP microdomain formation requires cross cell wall stabilization of domains and non-cell autonomous action of LOTR1
- Department of Plant Molecular Biology, University of Lausanne, Switzerland
- Electron Microscopy Facility, University of Lausanne, Switzerland
- Department of Applied Biological Chemistry, The University of Tokyo, Japan
Figures
Figure 1 with 1 supplement
Ectopic domain formation in lotr1.
(A) Schematic representation of endodermal maturation with stages of Casparian Strip (CS) development (green). Elongated endodermal cells focalize CASP1-GFP into aligned micro-domains which are …
Figure 1—figure supplement 1
Establishment of ectopic domain formations in lotr1.
(A) Localization of ectopic deposition of CASP1-GFP (green). Section view of z-stack through roots 5–10 cells after onset of protoxylem differentiation, cell wall stained with Calcofluor White …
Figure 2 with 1 supplement
CASP deposition is coordinated between neighboring endodermal cells.
(A) Double membrane phenotype visible with CASP1::CASP1-GFP during string-of-pearls stage of CS development, scale bars depict 10 µm. Red and gray arrows indicate separate membranes measured in (B). …
Figure 2—figure supplement 1
CASP1-GFP membrane coordination.
Double membrane phenotype of CASP1-GFP before and after ablation of one adjacent endodermal cell in early CSs. Cell walls (red) are stained with Propidium iodide (PI), yellow arrows indicate …
Figure 3 with 1 supplement
Endodermal neighbors are required for proper CSD establishment.
(A) Endodermal cell ablation leads to instability of the CSD in membranes facing dead cells. Endodermal cells were ablated 3–5 cells prior to onset of CASP1-GFP (white) expression and followed over …
Figure 3—figure supplement 1
Cell ablation impacts CS establishment.
(A) CASP1 promoter activity highlighted by nls-GFP (green) after ablation of all neighboring cells prior to onset of expression, PI (red) outlines cells and confirmed cell destruction. Scale bar = …
Figure 4 with 1 supplement
LOTR1 restricts CSD establishment through independent pathway.
(A) CASP1-GFP fluorescence of known CS mutants in wildtype and lotr1 background. Yellow arrows indicate parental CS mutant phenotype while blue arrows highlight lotr1 specific phenotypes; scale bar …
Figure 4—figure supplement 1
Investigation of putative post-translational processing in LOTR1.
(A) Lignin accumulation (magenta) in wildtype and sgn3 with CSDs labeled by CASP1-GFP (green). (B) Schematic view of LOTR1 with position of identified loss-of-function alleles indicated. (C) …
Figure 5 with 1 supplement
Stele-specific LOTR1 restricts CSD establishment.
(A) Mis-expression of LOTR1 in cortical cells causes dominant loss-of-function phenotypes. CASP1-GFP phenotypes of seedlings expressing LOTR1-complementation construct (used in Figure 4F) under …
Figure 5—figure supplement 1
Neprosin domain analysis in the HOLO family.
(A) Native LOTR1 expression is not dramatically changed in lines transformed with tissue-specific expression constructs used in Figure 5A. The data presented here are representative of three …
Figure 6
Speculative model of LOTR1 function.
The following model for LOTR1 function is simply one scenario that can plausibly integrate the data on LOTR1’s predicted activity, expression, subcellular localization, as well as phenotypes by …
Additional files
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Transparent reporting form
- https://cdn.elifesciences.org/articles/69602/elife-69602-transrepform1-v2.docx
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Source data 1
Source data files.
- https://cdn.elifesciences.org/articles/69602/elife-69602-supp1-v2.zip
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Supplementary file 1
Table S1.
- https://cdn.elifesciences.org/articles/69602/elife-69602-supp2-v2.xlsx
