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. 2017 Dec 18;7:17704. doi: 10.1038/s41598-017-18090-7

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© The Author(s) 2017

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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Validation of candidate gene. (a) Sequence variations in the promoter region of OsNRAMP5 between two parents. (b,c) Comparison of the relative expression level of OsNRAMP5 between 93–11 (grey) and CSSL-qGMN7.1 (white) in roots and shoots at the seedling stage (b), and in different organs at the booting stage (c). (d,e) Promoter activity analysis. The green fluorescent signals of GFP driven by the OsNRAMP5 promoter from 93–11 and PA64s (d), the relative expression level of GUS driven by the OsNRAMP5 promoters from 93–11 and PA64s (e). (f–h) Relative expression level of OsNRAMP5 and grain Mn and Cd concentrations in 93–11 and the overexpression (OX) lines grown in pots. *and **indicate a 5% and 1% significance level, respectively, according to the t test.