Extended Data Figure 10. Nutrient deficiency responses of suberisation in the xylem pole.

a) Expression of the suberin marker GPAT5 and the xylem pole pericycle marker XPP using a mCitrine-SYP122 or a 3mCherry-SYP122 plasma membrane-anchored reporter, respectively figure is representative of 5 independent lines. Plants were germinated for 5-days on ½ MS media, plates with low (10 μM) inorganic phosphate (low Pi), no zinc (-Zn) or no iron (-Fe) image is representative of 4 independent lines. b) Expression of the suberin marker GPAT5 and PHO1;H3 using a mCitrine-SYP122 or a NLS 3mScarlet reporter respectively in the suberised zone of Col-0 plants or plants expression the dominant cytokinin signaling repressor ARR10-GFP in all differentiating endodermal cells. Each image is representative of 8 independent lines. c) Expression of the suberin marker GPAT5 and PHO1;H3 using a mCitrine-SYP122 or a NLS 3Venus reporter respectively in the suberised zone of plants expressing estradiol-inducible (XVE) versions of the dominant cytokinin signaling repressor AHP6-GFP or the dominant ABA signaling repressor abi1-1 in all differentiated endodermal cells. Plants were grown for 5 days in presence of DMSO or 5 μM Estradiol (E2D), all treatments were repeated 3 times. Each image is representative of 5 independent lines. d) Expression of PHO1;H3 and the suberin marker GPAT5 in roots of 5-day-old Col-0 plants grown on ½ MS media, plates containing no iron (-Fe) no zinc (-Zn) with low (10 μM) inorganic phosphate (low Pi). Each image is representative of 9 independent lines. All treatments were repeated 3 times. e) Transcriptional analysis of PHO1;H3 expression in roots of 5-day-old Col-0 plants grown on ½ MS media, plates containing no iron (-Fe) no zinc (-Zn) with low (10 μM) inorganic phosphate (low Pi), in plants expression the dominant cytokinin signaling repressor ARR10-GFP in all differentiated endodermal cells or in plants with strongly reduced suberin deposition by expression of a suberin degrading enzyme (pCASP1::CDEF). Expression was normalized to that of UBQ10. Dots represent individual datapoints. Notice that for all stacked graphs there are 3 measurements per root (unsuberised zone (white), patchy zone (grey) and suberised zone (yellow)). Asterisks highlight unsuberised xylem pole endodermal cells. Boxplot centers show median. Individual letters show significantly different groups according to a post hoc Bonferroni-adjusted paired two-sided T-test. For more information on data plots see the statistics and reproducibility section. n represents independent biological samples. Scale bars represent 25 μm.