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. 2018 Sep 26;8:14412. doi: 10.1038/s41598-018-32819-y

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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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Analysis of cadmium sensitivity conferred by expression of cacao NRAMP genes in yeast. Yeast strain DY1457 carrying TcNRAMP1, 2, 3, 5, 5s (splice variant of TcNRAMP5), 6 or empty vector pDR195 (EV) were cultured on synthetic defined (SD)-Ura plates and liquid medium (b) in absence or presence of different concentration of CdCl₂. The plates were incubated at 30 °C for 3 days (a). For growth assay in liquid medium (b) the cells were grown at 0.001 OD₆₀₀ for 72 h at different concentrations of CdCl₂ as indicated in the figure. (c) Yeast strain DY1457 expressing TcNRAMP5, 6 or empty vector pDR195 (EV) were grown in SD-Ura liquid medium supplemented with 2 μM CdCl₂ for 72 h. The Cd²⁺ accumulation in the cells was determined by inductively coupled plasma mass spectrometry (iCAP™ Q ICP-MS). Data are means ± SD of three biological replicates.