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. 2018 Jun 1;69(16):4127–4139. doi: 10.1093/jxb/ery212

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© The Author(s) 2018. Published by Oxford University Press on behalf of the Society for Experimental Biology.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 3. — NADPH-oxidase-dependent H₂O₂ and nitrate reductase (NR)-dependent NO are essential for cold acclimation-induced MPK1/2 activation and cold tolerance. (A, D) REL in RBOH1- (A) and NR- (D) silenced plants. (B, E) NO accumulation in RBOH1-silenced plants (B) and H₂O₂ accumulation in the apoplast in NR-silenced plants (D). (C, F) MPK1/2 activation level in RBOH1- (C) and NR- (F) silenced plants. H₂O₂ at 10 mM and SNP at 500 μM were applied 12 h before the cold acclimation treatment. H₂O₂ and NO accumulation in leaves were visualized using the same protocols as shown in Fig. 2. Scale bars: 50 μm (B) and 1 μm (E). The numbers above the images in (C, F) indicate the relative intensity of MPK1/2 (the upper lane). REL was determined at 5 d, while the other parameters were assayed at 12 h after commencement of the cold stress treatment. Data are the means (±SD) of four biological replicates. Different letters indicate significant differences (P<0.05) according to Tukey’s test.