An evolutionarily young defense metabolite influences the root growth of plants via the ancient TOR signaling pathway
- University of Copenhagen, Denmark
- University of California, Davis, United States
Figures
Figure 1 with 1 supplement
3OHP reversibly inhibits root growth.
(A) 7-d-old seedlings grown on MS medium supplemented with a concentration gradient of 3OHP. (B) Quantification of root lengths of 7-d-old. Results are averages ± SE (n = 3–7; p<0.001). (C) …
Figure 1—figure supplement 1
Root inhibition is affected by endogenous GSL levels.
Results are least squared means ± SE over three independent experimental replicates with each experiment having an average of ten replicates per condition (n = 8–39). (A) Root growth for seedlings …
Figure 2 with 1 supplement
Root growth is not inhibited by all aliphatic GSLs.
(A) The aliphatic glucosinolate biosynthetic pathway, from the C3 3-methyl-sulphinyl-propyl (3MSP) to the secondary modified 3-hydroxyl-propyl (3OHP) and 2-propenyl (allyl/sinigrin). (B–C) Root …
Figure 2—figure supplement 1
Root lengths of 7 DAG Col-0 WT grown on MS media supplemented with 50 µM of the indicated GSL.
Progoitrin = R enantiomer of 2-hydroxybut3-enyl GSL and Epiprogoitrin is the S enantiomer. Results were obtained in two fully independent experiments and tested with ANOVA. LSmeans are shown with …
Figure 3 with 1 supplement
Conservation of 3OHP responsiveness suggests a evolutionally conserved target.
(A) Stylized phylogeny showing the phylogenetic relationship of the selected plants from the Brassicales family, branch lengths are not drawn to scale. (B–F) plants from the Brassicales family, …
Figure 3—figure supplement 1
Yeast response to 3OHP suggests a conserved target throughout eukaryotes.
Yeast growth in YPD media supplemented with none or increasing levels of 3OHP or Allyl. The hourly OD600 increase is plotted against each concentration of either Allyl or 3OHP. The least squared …
Figure 4
3OHP reduces root zone sizes.
(A) Diagrammatic organization of a root tip; the meristem zone from the QC to the first cell elongation; the elongation zone ends when first root hair appears (Dolan and Davies, 2004). (B) Meristem …
Figure 5 with 3 supplements
TOR over-activation amplifies 3OHP response.
(A) Root growth for low light grown seedlings. The seedlings were grown on MS medium without sucrose for 3 days, then transferred to the indicated media (Suc; sucrose). Multi-factorial ANOVA was …
Figure 5—figure supplement 1
Published TORox lines that did not display the TORox phenotype under our conditions.
Multi-factorial ANOVA was used to test the impact of Genotype (Col-0 v specific TORox lines) on root length. All experiments were combined in the model and experiment treated as a random effect. …
Figure 5—figure supplement 2
RAPTOR1 haplo-insufficiency does not affect 3OHP response.
(A) Root growth for heterozygous raptor1-2 and wildtype Col-0 seedlings grown on MS medium supplemented with or without 5 µM 3OHP. Multi-factorial ANOVA was used to test the impact of Genotype …
Figure 5—figure supplement 3
Loss of one of the two substrate-binding TORC-subunits affect 3OHP response.
(A) Root growth for raptor2-1 and wildtype Col-0 seedlings grown on MS medium supplemented with or without 5 µM 3OHP. Multi-factorial ANOVA was used to test the impact of Genotype (Col-0 v raptor2-1)…
Figure 6
3OHP dampens sugar-mediated meristem activation.
(A) Root growth for low light grown Col-0 wildtype seedlings. The seedlings were grown on MS medium without sucrose for 3 days, then transferred to the indicated media. Multi-factorial ANOVA was …
Figure 7 with 1 supplement
Pharmacological interaction of 3OHPGSL and the TOR inhibitor, AZD.
(A) Root lengths of 7-d-old Col-0 wildtype seedlings grown on MS medium with sucrose ±combinations of AZD and different concentrations of 3OHP. Results are least squared means ± SE over three …
Figure 7—figure supplement 1
Pharmacological interactiosn between 3OHPGSL and diverse TOR inhibitors.
(A) Root lengths of 7-d-old Col-0 wildtype seedlings grown on MS medium with sucrose ±the indicated mTOR or S6K inhibitors. Results are averages ± SE (n = 8–41). (B) Confocal images of 4–d-old …
Figure 8
Blocking autophagosome elongation amplifies the 3OHP response.
(A) Root growth for atg5-1 and wildtype Col-0 seedlings grown on MS medium supplemented with or without 5 µM 3OHP. Multi-factorial ANOVA was used to test the impact of Genotype (Col-0 v atg5-1), …
Additional files
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Supplementary file 1
Plant seeds used for in vitro root growth assays for the various plant species.
The source, common name, order, family, genus, species, and subspecies seeds are listed for the used plant species. As well as the plate size (cm × cm), and plating distance used for the individual response assays.
- https://doi.org/10.7554/eLife.29353.018
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Transparent reporting form
- https://doi.org/10.7554/eLife.29353.019
