Extended Data Fig. 2: Canonical ethylene signalling pathway inhibits periderm regeneration.

a,b,e, Cross sections of 18-day-old proPER49:erVenus (a) and proPBP1:erVenus (b) roots at 1 dai grown on MS plate supplemented without (Mock) or with 10 µM ACC (ACC) for one day after the injury. The proportion of cells at the wound site showing Venus signal intensities above the threshold was quantified at 1 dai in mock or ACC-treated 18-day-old proPER49:erVenus and proPBP1:erVenus roots (e). Two-tailed Wilcoxon rank-sum test was used (ns; P  ≥  0.05, **; P < 0.01). c,d,f, Cross sections of 14-day-old proPER15:erVenus (c) and proPBP1:erVenus (d) roots grown on MS plate supplemented without (Mock) or with 10 µM ACC (ACC) for two days. Venus signal intensities in periderm of proPER15:erVenus and proPBP1:erVenus without or with ACC treatment (f). Two-tailed Welch’s t-test (ns; P ≥ 0.05) was used. g, Cross sections of 16-day-old wild-type or ate1-2;ate2-1 roots grown on MS plate supplemented without (Mock) or with 10 µM ACC (ACC) for four days. h–k, Cross sections of 18-day-old proPER15:erVenus roots at 1 dai grown on MS plate supplemented without (Mock) or with 10 µM ACC (ACC) for one day after the injury. proPER15:erVenus seedlings in the wild-type background were introgressed into ein2-1 or etr1-3 mutants, and homozygous lines for proPER15:erVenus transgene and mutations were used (proPER15:erVenus;ein2-1, proPER15:erVenus;etr1-3). The proportion of cells at the wound site showing Venus signal intensities above the threshold was quantified in k at 1 dai in mock or ACC-treated 18-day-old proPER15:erVenus roots. Two-tailed Wilcoxon rank-sum test was used (ns; P  ≥  0.05, **; P < 0.01). l,m, Cross sections of 18-day-old proPER15:erVenus (l) or proPBP1:erVenus (m) roots at 1 dai grown without (Mock) or with 30 ppm ethylene (Ethylene) for one day after the injury. n, Cross sections of 19-day-old wild-type roots at 4 dai grown without (Mock) or with 30 ppm ethylene (Ethylene) for four days after the injury. o, The proportion of cells at the wound site showing Venus signal intensities above the threshold was quantified at 1 dai in mock or ethylene-treated 18-day-old proPER15:erVenus (left) and proPBP1:erVenus (middle) roots. Two-tailed Wilcoxon rank-sum test was used (**; P < 0.01). The density of suberized cells at the wound site was quantified in mock or ethylene-treated 19-day-old wild-type at 4 dai (right). Two-tailed Wilcoxon rank-sum test was used (*; P < 0.05). n indicates the number of examined cross sections. Venus signal intensities in a–d, h–j, l–n and intensity of suberin staining with Fluorol yellow in g,n are shown according to the colour map on the right. Fractions on the panel indicates the proportion of cross sections showing the similar subrization as in the images. White, SR2200 (cell wall). Scale bars: 50 µm.

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