1. Rhamnose-Containing Cell Wall Polymers Suppress Helical Plant Growth Independently of Microtubule Orientation 
Adam M. Saffer, Nicholas C. Carpita, Vivian F. Irish 
Current Biology 
Volume 27, Issue 15, Pages e4 (August 2017)
DOI: /j.cub
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2. Current Biology 2017 27, 2248-2259.e4DOI: (10.1016/j.cub.2017.06.032)
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3. Figure 1 dq Petal Epidermal Cells Twist into a Left-Handed Helix
Images of flowers and petals from mature stage 14 Ler wild-type (A–E) and dq mutant (F–J) plants. SEM of adaxial epidermal cells from the distal petal blade region viewed from the side (B and G) or top (C and H), abaxial epidermal cells from the distal petal blade region (D and I), and adaxial epidermal cells from the petal claw (E and J). Scale bars, 1 mm in (A) and (F) and 10 μm in (B)–(E) and (G)–(J).
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4. Figure 2 rhm1 Mutants Have Left-Handed Helical Petals
(A) Cartoon of RHM1 showing the locations of the rhm1-1 nonsense mutation, rhm1-3 and rhm1-2 missense mutations, and the rhm1-4 T-DNA insertion site.
(B–D) Expression of a rescuing genomic RHM1::GFP construct. (B) Adaxial petal epidermal cells. (C) Abaxial petal epidermal cells. (D) Optical projection of a z stack of images showing a profile view of a petal with predominantly epidermal expression. ad, adaxial epidermis; ab, abaxial epidermis; m, mesophyll.
(E–Q) Images of mature stage 14 flowers. (E) Ler WT. (F) Col WT. (G) rhm1-3 mutant. (H) rhm1-3 mutant carrying a transgene containing the RHM1 genomic locus. (I) rhm1-1 mutant. (J) rhm1-2 mutant. (K) rhm1-4 mutant. (L) rhm1-3/rhm1-1 heterozygote. (M) rhm1-3/rhm1-2 heterozygote. (N) rhm1-3/rhm1-4 heterozygote. (O) rhm1-1/rhm1-2 heterozygote. (P) rhm1-1/rhm1-4 heterozygote. (Q) rhm1-2/rhm1-4 heterozygote. WT, wild-type.
Scale bars, 20 μm for (B)–(D) and 1 mm for (E)–(Q). See also Figure S1 and Table S1.
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5. Figure 3 rhm1 Is Required for Adaxial Petal Epidermal Cell Expansion
(A) Developmental progression of adaxial petal epidermal cells in Ler wild-type and rhm1-3 mutants. Wild-type (WT) and rhm1-3 images were each taken from a single inflorescence.
(B–D) SEM images of adaxial petal epidermal cells from stage 14 flowers. (B) Col WT. (C) rhm1-1. (D) rhm1-2. Scale bars, 10 μm for all images.
(E) Mean cone height of stage 14 petals from at least 18 different plants per genotype, with a minimum of ten cells measured per plant. Error bars indicate SD. ∗p < 0.0001, compared to the corresponding wild-type (t test).
See also Figure S2.
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6. Figure 4 rhm1 Mutants Exhibit Helical Root Growth
Roots were grown for 7 days on vertically oriented hard agar plates.
(A) The average orientation of roots on each plate was determined, with vertical roots being 0°, angles < 0° representing left-handed growth, and angles > 0° representing right-handed growth. A minimum of ten plates grown on 4 different days were assayed for each genotype and the average orientation ± SD is shown.∗p < 0.005, and ∗∗p < for comparison of rhm1 mutant plants to the corresponding plants with wild-type RHM1 by paired t tests. WT, wild-type.
(B) Higher-magnification images show the arrangement of root epidermal cell files.
Scale bars, 1 cm for plate images in (A) and 100 μm for (B). See also Table S2.
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7. Figure 5
Rhamnose-Containing Cell Wall Polymers Suppress Helical Root Growth
Roots were grown for 7 days on vertically oriented hard agar plates.
(A) The average orientation of roots on each plate was determined, with vertical roots being 0°, angles < 0° representing left-handed growth, and angles > 0° representing right-handed growth. At least ten plates grown on 4 different days were assayed for each genotype and the average orientation ± SD is shown. ∗p < 0.005, and ∗∗p < for comparison of mutant plants to the wild-type by paired t tests. WT, wild-type.
(B) Higher-magnification images show the arrangement of root epidermal cell files.
Scale bars, 1 cm for plate images in (A) and 100 μm for (B). See also Figure S3 and Table S2.
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8. Figure 6 rhm1 Mutants Do Not Affect Root Microtubule Orientation
(A) Root length and growth angle were assayed in seedlings grown for 7 days in the presence of propyzamide. Root growth angles < 0° represent left-handed growth, and angles > 0° represent right-handed growth. At least four plates were assayed for each data point. WT, wild-type.
(B–H) Immunostaining of tubulin in epidermal cells from the root elongation zone. (B–G) A single representative cell is shown for each genotype. (H) Average microtubule orientation was determined for each cell. 90° represents transverse microtubules, angles > 90° represent right-handed microtubules, and angles < 90° represent left-handed microtubules. Average angle ± SD is shown. The frequency of each average orientation in 4° bins is plotted. At least 195 cells from at least 72 different roots were analyzed for each genotype.
Error bars indicate SD. Scale bars, 10 μm for all images. MT, microtubule. See also Figure S5.
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