Fig. 3: The root tips of arh1-2 fei1-C fei2-C show defects in CCW.

a The amount of cellulose in the cell wall of the root tips of Col-0 and the triple mutant was quantified. The total cell wall was presented as alcohol-insoluble residues (AIR). b D-Galacturonic acid monohydrate was measured in cell wall (AIR) of Col-0 and the triple mutant root tips by GC-MS. Ruthenium red stained root tips from four-day-old Col-0 (c) and the triple mutant (d) seedlings. e Relative staining intensity for the root tips as represented in (c), (d) in a 200 µm × 100 µm area (as shown in c). f–h, Fluorol yellow (FY) stained root tips from seedlings of two-day-old Col-0, arh1-2 fei1-C fei2-C, and gso1 gso2 mutants. i–t Transmission electron microscope (TEM) images showing CCW of the outermost lateral root cap cells (100 μm from the root tip) (i-p) and the outermost epidermal cells in the elongation zone (400 μm from the root tip) (q–t) of two-day-old Col-0, arh1-2 fei1-C fei2-C, gso1 gso2, and kcs7 kcs15 kcs20 kcs21 quadrupole mutants. LRC represents lateral root cap. Data are the means ± SD of three biological replicates (a, b). Each circle represents a measurement of a biologically independent sample (a, b) or an individual root tip (e). “n” represents the number of roots analyzed in the experiment (e). Three biological replicates were carried out for ruthenium red staining and YF staining experiments, while ten biological replicates were carried out for TEM analyses. Scale bars represent 50 µm in (c, d, f–h), 3 µm in (i–l), and 300 nm in (m–t). Statistical significance was determined by two-side and unpaired t-test, without making any adjustments for multiple comparisons (P < 0.01).