Plants monitor the integrity of their barrier by sensing gas diffusion

Summary

Plant materials and growth conditionsCol-0 was used as the wild type unless stated otherwise. ein2-1, etr1-3, ate1-2;ate2-1, prt6-5, proWOX4-erYFP, proPXY:GUS, proPXY:erVenus and 35S:EIN3-GFP have been described previously10,13,27,37,38,39,40. ate1-2;ate2-1 mutants were reciprocally backcrossed into Col-0 for three generations and transfer DNA insertions were confirmed by genotyping to obtain double-homozygous mutants.Seeds were sown on half-strength MS (Duchefa) plates supplemented with 0.05% MES (Duchefa), 1% agar (Duchefa) and 1% sucrose (Duchefa), pH 5.8. MS medium also contained vitamins for all experiments except for ethylene measurement from roots. After incubation at 4 °C for more than 2 days, the plates were moved to the growth chamber (22 °C; 16 h light, 8 h dark). The day that the plates were placed in the growth chamber is defined as day 0. The seedlings were moved to the new MS agar plate around day 7; for the experiment in shoots, the seedlings were transferred to soil 1 or 2 weeks after germination and were grown in a greenhouse (16 h light, 8 h dark) unless stated otherwise. For examining normal periderm, we used 14- or 16-day-old seedlings as periderm is established at that age. For assessing regeneration, we usually used 17- to 21-day-old seedlings.The Arabidopsis Genome Initiative locus codes for the genes are as follows: ATE1, AT5G05700; ATE2, AT3G11240; EBF1, AT2G25490; EIN2, AT5G03280; EIN3, AT3G20770; ETR1, AT1G66340; PBP1, AT3G16420; PCO1, AT5G15120; PCO2, AT5G39890; PER15, AT2G18150; PER49, AT4G36430; PRT6, AT5G02310; PXY, AT5G61480; RPS5A, AT3G11940; WOX4, AT1G46480.CloningFrom the Col-0 genome, the 643-base-pair (bp) EBF1 3′ UTR with its downstream 72-bp sequence (from +2780 to +3495) was amplified by PCR using H122_EBF1_URT_F and H123_EBF1_URT_R primers18 (Supplementary Table 1), which have a 25-bp overlap with the erVenus and attL2 sequence, respectively (EBF1 3′ UTR). erVenus/pDONR plasmid (30 ng) was amplified with the EBF1 3′ UTR PCR ...