Extended Data Fig. 10: Hypoxia and ethylene signalling play a minor role in the formation of phellem-like layer in wounded inflorescence stems.

a, Cross sections of proPER49:erVenus, proPBP1:erVenus, proAT3G26450:erVenus, and proAT1G14120:erVenus inflorescence stems one or three days after injury. b, Cross sections of proPCO1:erVenus and proPCO2:erVenus inflorescence stems. The inflorescence stems without injury (Intact) or two days after the injury. proPCO1:erVenus and proPCO2:erVenus signals were broadly detected in their inflorescence stems with higher expression in the vascular bundle. c, Cross sections of wild-type and ate1-2;ate2-1 inflorescence stems four days after injury. d, Quantification of concentration of ethylene emitted from inflorescence stems. The wild-type inflorescence stems were not (Control) or were (Wounded) injured along the longitudinal axis. The ethylene concentration was measured 3, 6, and 24 h after the injury. Higher ethylene concentration was always detected from wounded inflorescence stems at all time points in each replicate. Two-tailed Welch’s t-test was used (ns; P  ≥  0.05, *; P < 0.05). e,h,i, Cross sections of RPS5A:erVenus and RPS5A:erVenus-EBF1UTR intact inflorescence stems (Intact) or at 2 dai treated without (Cut) or with (Cut + Vaseline) Vaseline. Venus signal intensities in the cortical cells or in the cortical cells near the wound were measured in RPS5A:erVenus and RPS5A:erVenus-EBF1UTR intact or wounded inflorescence stems. Two-tailed Welch’s t-test or two-tailed Wilcoxon rank-sum test was used (ns; P ≥ 0.05, **; P < 0.01) for h and i, respectively. f,j, Cross sections of inflorescence stems of 21-day-old proPER15:erVenus seedlings at 1 dai grown on MS plate supplemented without (Mock) or with 10 µM ACC (ACC) for one day after the injury. Quantification of proPER15:erVenus signal intensities at 1 dai. Two-tailed Welch’s t-test was used (**; P < 0.01). g, Cross sections of wild-type, ein2-1 and etr1-3 inflorescence stems two or four days after the injury. The region covered with suberized cells at the wound site was quantified. One-way ANOVA test followed by Dwass-Steel-Critchlow-Fligner pairwise comparisons was used (same characters indicate statistically not-significant differences between two groups; P > 0.05). n indicates the number of repeats in d or the number of examined cross sections in g–j. Venus signal intensities in a,b,e,f and intensity of suberin staining with Fluorol yellow in c,g are shown according to the colour map on the right. White, SR2200 (cell wall). Fractions on the panels indicate the proportion of cross-sections showing the similar expression or suberized cell formation as in the images. Scale bars, 50 µm.

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