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. 2017 Sep 8;7:11051. doi: 10.1038/s41598-017-11428-1

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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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Longitudinal sections of a Medicago 20 dpi nodule showing the distribution of cells infected with Micromonospora ML01-gfp and Sinorhizobium Rm1021-mCh captured by CLSM. (a) Detail of a nodule tip captured with the green channel for the localization of Micromonospora. Arrows indicate fluorescence emitted by Micromonospora to differentiate from autofluorescence emitted by the plant. (b) Detail of a nodule tip captured with the red channel for the localization of Mesorhizobium Rm1021-mCh. (c) Composite image showing the distribution of green and red fluorescence. (d) Bacteroids occupying several plant cells, small circles show areas occupied by both bacteria. (e) Micromonospora hyphae inside host plant cells. The white circle in image 7c shows the area where images d and e were captured. Bars: 75 µm (a); 30 µm (b–e). dpi, days post inoculation; C, cortex; BZ, infection zone; AF, autofluorescence; b, bacteroids; m, Micromonospora.