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. 2021 Jan 8;11:159. doi: 10.1038/s41598-020-80377-z

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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STEM analyses of a transverse section of a rice root exposed to enhanced biochar. (a) Bright field (BF) image of a microbe attached to the root epidermis. The two rectangular regions of interest are where EDS/EELS analyses were performed. The EDS spectra show a concentration of Si with small amounts of Fe at the surface layer of the microbe (Region 1), and significant concentrations of Si, Fe, and P at the microbe-root interface (Region 2). The Fe L-edge EEL spectrum of region 2 is shown inset in (a) where the edge shape is typical of Fe₃O₄. (b) High-angle annular dark field (HAADF) image of the intracellular agglomerate of the root. The EDS spectrum from the region arrowed is shown inset where a high concentration of Fe is detected. (c) A magnified BF image of the agglomerate showing matrices containing an iron-rich core with a diameter of around 5 nm. (d) BF image of an intracellular cluster of particles showing high concentrations of Si and Fe detected by EDS analysis. (e) Detailed view of the boxed region in (d) showing the needle-like particles, with the inset showing the Fe L-edge EEL spectrum typical for iron minerals with mixed oxidation states (Fe(II)/Fe(III)).