KATANIN-dependent mechanical properties of the stigmatic cell wall mediate the pollen tube path in Arabidopsis
- Laboratoire de Reproduction et Développement des Plantes, Université de Lyon, ENS de Lyon, UCBL, INRAE, France
- Institut Jean-Pierre Bourgin, INRAE, AgroParisTech, Université Paris-Saclay, France
Figures
Figure 1
CMT organisation during papilla cell development.
(A) Flower development of A. thaliana from developmental stages 12 to 15. Scale bar, 500 μm. (B) Upper view of the stigma during development by SEM. Scale bar, 50 μm. (C) Confocal images of papilla …
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Figure 1—source data 1
Source data for CMT organisation shown in Figure 1D.
- https://cdn.elifesciences.org/articles/57282/elife-57282-fig1-data1-v1.xlsx
Figure 2
Pollen tube growth behaviour on papillae during development.
(A) SEM images of Col-0 papillae pollinated with Col-0 pollen, one hour post pollination, from stages 12 to 15. Scale bar, 5 μm. (B) Quantification of the number of turns (T) made by the pollen tube …
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Figure 2—source data 1
Source data for the number of turns made by the pollen tube on papillae from stage 12 to 15 shown in Figure 2B.
- https://cdn.elifesciences.org/articles/57282/elife-57282-fig2-data1-v1.xlsx
Figure 3 with 1 supplement
CMT organisation following pollen tube growth.
(A) Time-lapse imaging of Col-0 pollen tube growth within a papilla cell expressing MAP65.1-citrine. Pollen outline and pollen tube path were highlighted in magenta for a better visualisation. N = 5 …
Figure 3—figure supplement 1
Confocal image of the effect of mechanical stimulation on CMT organisation.
Coverslide was pressed manually leading to a destabilization of the CMT array of papillae expressing MAP65.1-citrine. Scale bar, 30 μm.
Figure 4
Local oryzalin application on Col-0 stigmas promotes MT destabilization and induces Col-0 pollen tube coils.
(A) Col-0 papilla cells expressing MAP65.1-citrine after 4 hr of DMSO (top) or oryzalin (bottom) local treatment. (B) SEM images of DMSO- (top) or oryzalin-treated (bottom) Col-0 stigmas pollinated …
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Figure 4—source data 1
Source data for the quantification of the number of turns made by Col-0 pollen tubes on drug-treated and control papillae shown in Figure 4C.
- https://cdn.elifesciences.org/articles/57282/elife-57282-fig4-data1-v1.xlsx
Figure 5 with 5 supplements
Effect of CMT organisation on pollen tube path.
(A) Confocal images of papilla cells expressing MAP65.1-citrine in Col-0 and ktn1-5 at stage 13. Scale bars, 5 μm. (B) SEM images of Col-0 and ktn1-5 papillae pollinated with Col-0 pollen grains. …
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Figure 5—source data 1
Source data for the quantification of CMT anisotropy of Col-0 and ktn1-5 papilla cells at stage 13 shown in Figure 5C and the quantification of the number of turns made by Col-0 pollen tubes on ktn1-5 and Col-0 papillae shown in Figure 5D.
- https://cdn.elifesciences.org/articles/57282/elife-57282-fig5-data1-v1.xlsx
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Figure 5—source data 2
Table of cell wall mutants analysed.
- https://cdn.elifesciences.org/articles/57282/elife-57282-fig5-data2-v1.docx
Figure 5—figure supplement 1
Col-0 pollen tube behaviour on Col-0 and ktn1-5 papillae at stage 13.
(A and B) Top views of Col-0 (A) and ktn1-5 (B) stigmas pollinated with Col-0 pollen grains. Scale bars, 20 µm. (C and D) Magnification of pollinated papilla cells. (C) Col-0 pollen tube grows …
Figure 5—figure supplement 2
ktn1-5 is impaired in female receptivity and fertility.
In all crosses, the female partner is written first. (A) Stigma receptivity: percentage of Col-0 hydrated pollen grains on Col-0 and ktn1-5 stigmas (N = 10 stigmas for each genotype, with 972 pollen …
Figure 5—figure supplement 3
Quantification of the number of coils made by Col-0 pollen tubes on papillae from cell-wall mutants at stage 13.
(A) xxt1 xxt2 x Col-0. N(Col-0)=14 stigmas, n(Col-0)=116 papillae, N(xxt1 xxt2)=12 stigmas, n(xxt1 xxt2)=116 papillae. (B) prc1 x Col-0. N(Col-0)=14 stigmas, n(Col-0)=115 papillae, N(prc1)=12 …
Figure 5—figure supplement 4
Cell wall composition is altered in katanin1 and different from xxt1 xxt2.
(A) Relative abundance of monosaccharides in the cell wall extracted from pistils of Col-0, ktn1, xxt1 xxt2. (B) MALDI-TOF analysis of xyloglucan from dissected stigmas of Col-0 and ktn1 (xxt1 xxt2 …
Figure 5—figure supplement 5
Size of the papilla cells of Col-0 and cell wall mutants.
(A) Papilla cell length of Col-0 at stage 13 and stage 15. (B) Papilla cell width of Col-0 at stage 13 and stage 15. (C) Papilla cell length of Col-0 and ktn1-5 at stage 13. (D) Papilla cell width …
Figure 6 with 1 supplement
Mechanical properties of papilla cell walls.
(A) Location of a Col-0 pollen tube in the cell-wall of Col-0 and ktn1-5 papillae by TEM. pt = pollen tube, C = stigma cuticle, sCW = stigma cell wall, sp = stigma papilla. Scale bar, 1 μm. (B) …
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Figure 6—source data 1
Source data for defining the mechanical properties of the papilla cell walls shown in Figure 6D, Figure 6F, Figure 6G, Figure 6H.
- https://cdn.elifesciences.org/articles/57282/elife-57282-fig6-data1-v1.xlsx
Figure 6—figure supplement 1
Ultrastructural features of Col-0 and ktn1-5 papilla cells.
(A) TEM images of non-pollinated Col-0 and ktn1-5 papillae. Scale bar, 5 μm. (B) Both Col-0 and ktn1-5 papillae show a similar two-layered cell wall (*) surrounded by an electron dense cuticle …
Figure 7
Cellulose microfibril organisation in Col-0 stage 13, ktn1-5, Col-0 stage 15 and xxt1 xxt2 papillae.
(A) 3D-projections of z-stack images of Col-0 stage 13, ktn1-5, Col-0 stage 15 and xxt1 xxt2 papilla cells stained with Direct Red 23 dye. Scale bars, 10 µm. (B) Plot profiles of the fluorescence …
Author response image 1
Stiffness differences between Col-0 and ktn1-5 papillae at stage 13.
Young’s modulus values of the papilla cell wall for Col-0 at stage 13 (N = 7 stigmas, n = 33 papillae) and ktn1-5 (N = 7 stigmas, n = 9 papillae). Statistical differences were calculated using a …
Author response image 2
Col-0 pollen tube trajectory after attachment of the pollen to different positions on the Col-0 papilla at stage 13.
Pollen tube trajectory is not dependent on the position where pollen has landed.
Author response image 3
Col-0 papilla cells expressing MAP65.
1-citrine after 6 or 4 hours of oryzalin local treatment. CMT depolymerization is complete and homogeneous after 4 hours of treatment at 888µg/mL. At 404 µg/mL and 666 µg/mL, CMTs are not completely …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent Arabidopsis thaliana | ktn1-5 | Lin et al., 2013 | AT1G80350 | |
| Genetic reagent Arabidopsis thaliana | xxt1 xxt2 | Cavalier et al., 2008 | AT3G62720 AT4G02500 | |
| Genetic reagent Arabidopsis thaliana | prc1.1 | Fagard et al., 2000 | AT5G64740 | |
| Genetic reagent Arabidopsis thaliana | qua2.1 | Mouille et al., 2007 | AT1G78240 | |
| Genetic reagent Arabidopsis thaliana | xyl1.4 | Sampedro et al., 2010 | AT1G68560 | |
| Genetic reagent Arabidopsis thaliana | kor1.1 | Nicol et al., 1998 | AT5G49720 | |
| Genetic reagent Arabidopsis thaliana | any1 | Fujita et al., 2013 | AT4G32410 | |
| Genetic reagent Arabidopsis thaliana | pSLR1::MAP65.1-citrine | This paper | Transgenic line expressing MAP65.1-citrine in stigmas, request to RDP laboratory, ENS Lyon, France | |
| Genetic reagent Arabidopsis thaliana | pSLR1::LTI6B-GFP | This paper | Transgenic line expressing LTI6B-GFP in stigmas used in Figure 6, request to RDP laboratory, ENS Lyon, France | |
| Genetic reagent Arabidopsis thaliana | pACT11::RFP | This paper | Transgenic line expressing RFP under the ACTIN11 promoter used to visualize pollen and pollen tubes in Figure 3 and Figure 6, request to RDP laboratory, ENS Lyon, France | |
| Chemical compound, drug | Oryzalin | Chemical Service, Supelco | 36182 | 833 µg/mL |
| Chemical compound, drug | RedDirect23 | Sigma-Aldrich | 212490 | 0.02% (w/v) |
| Chemical compound, drug | Adigor | Syngenta | 2.5% (v/v in water) | |
| Software, algorithm | ImageJ | RRID:SCR_003070 | https://imagej.net | |
| Software, algorithm | FibrilTool | Boudaoud et al., 2014 | RRID:SCR_016773 | https://biii.eu/fibriltool |
| Software, algorithm | Rstudio | RStudio Team, 2015 | RRID:SCR_000432 | https://rstudio.com/ |
