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. 2018 Apr 10;9:1376. doi: 10.1038/s41467-018-03832-6

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — NIGT1 proteins redundantly regulate NRT2.1 expression. a Phylogenic analysis of Arabidopsis and rice NIGT1/HHO proteins. Arabidopsis and rice proteins are shown in black and blue, respectively. Numbers indicate bootstrap values of 1000 iterations. b Time-dependent expression patterns of NIGT1-clade genes after 10 mM KNO₃ supply. c Concentration-dependent induction by nitrate. RNA was prepared from ammonium-grown seedlings before treatment and after treatment with 0, 0.1, 0.5, 1 and 10 mM KNO₃ for 1 h. A 10 mM KCl treatment was used as a control. d Nitrate-specific induction of NIGT1-clade genes. Ammonium-grown seedlings were treated for 1 h with 10 mM of each chemical, except for trans-zeatin (5 µM). e Repression of the NRT2.1 promoter by NIGT1 proteins. Protoplasts co-transfected with NIGT1 expression vectors or the empty vector (none) together with the reporter plasmid containing the LUC gene under the control of the NRT2.1 promoter were incubated in the presence of 1 mM KCl or KNO₃. LUC activity was normalised to GUS activity from the co-transfected UBQ10-GUS plasmid. Data are means ± s.d. of three biological replicates. f Transcript levels of NRT2.1 in the nigt1 quadruple mutants (Q-1 and Q-2) and NIGT1.1-OX plants after 10 mM KNO₃ supply. Values are normalised to those of UBQ10, and are means of biological triplicates ± s.d. (b, c, d, f)