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. 2018 May 16;8:7614. doi: 10.1038/s41598-018-25910-x

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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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Changes in GmBEHL1 expression affect nodulation. (a–c) GmBEHL1 overexpression suppresses nodulation. (a) qRT-PCR analysis of transgenic hairy roots expressing 35S:GmBEHL1. The relative expression levels of GmBEHL1 in hairy roots transformed with empty vector 1 (EV1) or 35S:GmBEHL1. The expression levels were normalized against the geometric mean of soybean GmELF1b for GmBEHL1. Student’s t-test was performed (***p < 0.001, n = 25). (b) Quantitative analysis of the nodule number per hairy root expressing EV1 and 35S:GmBEHL1 (n = 25). All values are the means ± SDs from more than three independent experiments. Asterisks represent statistically significant differences (Student’s t-test, ***p < 0.001). (c) Nodules of individual hairy roots expressing EV1 and 35S:GmBEHL1 at 28 DAI. Bar = 3 mm. (d–f) Knocking down GmBEHL1 promotes nodulation. (d) Expression analysis of transgenic hairy roots expressing empty vector 2 (EV2) and amiR-GmBEHL1. The expression levels of GmBEHL1 were normalized against the geometric mean of soybean GmELF1b. Student’s t-test was performed (n = 25, ***p < 0.001). (e) Quantitative data showing the nodules per hairy root for EV2 and the vector harboring amiR-GmBEHL1 (n = 25). The nodule numbers are the means ± SDs from more than three independent experiments. Asterisks represent statistically significant differences (Student’s t-test; ***p < 0.001). (f) Nodules of individual hairy roots expressing EV2 and amiR-GmBEHL1 at 28 DAI. Bar = 3 mm.