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. 2014 Sep 16;3:e03115. doi: 10.7554/eLife.03115

Figure 1. SGN3 receptor-like kinase is important to establish a functional endodermal barrier.

(A) Lack of endodermal diffusion barrier in sgn3-3 visualized by presence of propidium iodide (PI) in stele. (B) Diagram of the SGN3 protein showing the different domains, T-DNA insertion lines (indicated with triangles) and the ethyl methanesulfonate (EMS)-induced mutations (see also Figure 1—figure supplement 2). (C) Surface view of Casparian strip, visualized by autofluorescence after clearing. Note discontinuous lignin deposition in sgn3-3. Pictures are maximum projections of confocal z-stacks. Arrowheads indicate discontinuities in sgn3. Spiral-like signal in WT is from deeper-lying xylem vessel. (D) Fluorol yellow staining of suberin lamellae deposition in sgn3-3 and WT. Pictures are overlays of transmitted light image (gray) with fluorescent signal from suberin dye (yellow). (E) Occurrence of suberin deposition along the root is not altered in sgn3-3. Suberin lamellae deposition was quantified considering three different zones: non-suberized zone, zone of patchy suberization, and zone of continuous suberization (n = 5, one representative experiment presented). (F) Surface view of CSD network visualized with CASP1-GFP expressed under CASP1 promoter showing the net-like structure with discontinuities in sgn3-3. Projections as in C. (G) Absence of a lateral diffusion barrier in sgn3-3 visualized with plasma membrane marker line CASP1::mCherry-SYP122 (intensity color coded). Confocal pictures were taken at the surface of an endodermal cell. Note the mutually exclusive localization with the CSD marker CASP1-GFP (green). Two right images are magnification of the two leftmost images. (H) Localization of the outer marker PDR6-Venus expressed under CASP1 promoter is still polar in sgn3-3. Pictures are median longitudinal sections of endodermal cells. Scale bars: A, C, F, G, H = 20 μm; D = 50 μm. ep, epidermis; co, cortex; en, endodermis; st, stele; LRR, Leucine-rich repeat.

DOI: http://dx.doi.org/10.7554/eLife.03115.003

Figure 1—source data 1. Detail of SGN3 T-DNA and EMS mutants.
Detail of SGN3 mutations. sgn3-1 EMS mutant was lost during the screen and is not available anymore.
DOI: http://dx.doi.org/10.7554/eLife.03115.004
elife03115s001.xls (17.5KB, xls)
DOI: 10.7554/eLife.03115.004

Figure 1.

Figure 1—figure supplement 1. Both Casparian strip domain and Casparian strip but not the suberin are affected in sgn3.

Figure 1—figure supplement 1.

(A) Lack of endodermal diffusion barrier in sgn3-3 and sgn3-4 visualized by free diffusion of propidium iodide (PI) into stele. (B) Surface view of Casparian strips by autofluorescence after clearing shows discontinuous lignin deposition in sgn3-3 and sgn3-4. Pictures are maximum projections of confocal z-stacks. (C) Fluorol yellow staining in WT, sgn3-3, and sgn3-4. (D) Suberin deposition along the root is not altered in sgn3-3 and sgn3-4. Suberin lamellae deposition was quantified as in Figure 1E (n = 5, one representative experiment shown). (E) Surface view of CSD network visualized with CASP1-GFP, CASP2-GFP, CASP3-GFP, CASP4-mCherry, and CASP5-GFP under CASP1 promoter showing the discontinuous net-like structures in sgn3-3. Projections as in B. Scale bars: A, B, E, 20 μm; C, 50 μm. ep, epidermis; co, cortex; en, endodermis; st, stele.
DOI: http://dx.doi.org/10.7554/eLife.03115.005
Figure 1—figure supplement 2. Diagram of SGN3 genomic DNA with T-DNA and EMS mutants.

Figure 1—figure supplement 2.

Schematic of SGN3 genomic region showing the insertions sites of the three T-DNA mutants and the EMS mutants. sgn3-2 was found in a forward genetic, GUS-based screen (Alassimone et al., unpublished), sgn3-5 to sgn3-18 were found in additional forward genetic screen (Kalmbach et al., unpublished). sgn3-3, sgn3-4, and sgn3-19 are T-DNA insertion mutants in the first exon of SGN3.
DOI: http://dx.doi.org/10.7554/eLife.03115.006