Extended Data Fig. 3: Ethylene reporter line validation and its expression after wounding.

a,e, Cross sections of 14-day-old RPS5A:erVenus and RPS5A:erVenus-EBF1UTR roots (a) and Venus signal intensities in distal phloem parenchyma (e). RPS5A:erVenus signals were stronger than in RPS5A:erVenus-EBF1UTR, especially in the periderm and phloem parenchyma indicating high ethylene signalling in these outer tissue types. b,f,g, Cross sections of 16-day-old RPS5A:erVenus and RPS5A:erVenus-EBF1UTR grown on MS plate (Mock), MS plate supplemented with 10 µM ACC (ACC) for two days, 10 µM AVG (AVG) or 0.5 mM AgNO3 (AgNO3) for four days (b). The normalized Venus signal intensities in vascular cambium (f,g). c,h,i, Confocal microscopy images of RPS5A:erVenus and RPS5A:erVenus-EBF1UTR root tip grown on MS plate supplemented without (Mock) or with 10 µM ACC (ACC) or 0.5 mM AgNO3 (AgNO3) for 24 h (c). Venus signal intensities in the epidermis (h,i). d,j,k, Cross sections of 19-day-old RPS5A:erVenus and RPS5A:erVenus-EBF1UTR roots right after (left) or two days after (right) injury (d). While RPS5A:erVenus signal was not changed after injury, RPS5A:erVenus-EBF1UTR signals were increased near the wound site, indicating reduced ethylene signalling. Venus signal intensities in distal phloem parenchyma near the wound (j,k). Two-tailed Welch’s t-test (ns; P ≥ 0.05, **; P < 0.01) was used for e,j,k. Kruskal-Wallis test (P < 0.01) followed by Dwass-Steel-Critchlow-Fligner pairwise comparisons (f,g) or Pairwise Two-tailed Welch’s t-test for multiple comparisons with holm correction (h,i) was used (different characters indicate statistically significant differences between two groups; P < 0.05). n indicates the number of examined cross sections or root tips. Venus signal intensities are shown according to the colour map on the right. White, SR2200 (cell wall). Scale bars: 50 µm.

Source Data