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. 2020 Sep 28;10:15887. doi: 10.1038/s41598-020-72429-1

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© The Author(s) 2020

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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The FliC (N87K) substitution alters flagellar shape and swimming mode. (a) Left: fliC gene and protein sequence comparison. Right: Crystal structure of FliC (PDB: 1IO1). The red structure represents N87 residue. (b) Electron micrographs of SHU102 [fliC (N87K):: fliC,], HCB1336/pYS10, and HCB1336/pSHU61. Scale bars, 2 μm. (c) Histograms of the flagellar pitch observed in SHU102 (top), HCB1336/pYS10 (middle), and HCB1336/pSHU61 (bottom) cells with peaks (± SDs) at 2.5 ± 0.2 μm (n = 27), 2.7 ± 0.3 μm (n = 42), and 1.4 ± 0.1 μm (n = 46), respectively. (d) Histogram of the helix radius observed in SHU102 (top), HCB1336/pYS10 (middle), and HCB1336/pSHU61 (bottom) cells with peaks (± SDs) at 0.20 ± 0.04 μm (n = 27), 0.22 ± 0.06 μm (n = 44), and 0.14 ± 0.03 μm (n = 46), respectively. (e) Cell motility on 0.25% (wt/vol) soft-agar plates after 7 h at 30 ºC. Scale bar, 1 cm. (f) Swimming traces at 150 ms intervals for 15 s. The intermittent color code indicates the time course from red to blue. Area, 68.6 × 85.9 μm. (g) Swimming speed histograms of SHU102 (left), HCB1336/pYS10 (middle), and HCB1336/pSHU61 (right) cells with peaks (± SD) at 26.3 ± 6.0 μm s⁻¹ (n = 45), 21.7 ± 4.5 μm s⁻¹ (n = 53), and 12.0 ± 4.2 μm s⁻¹ (n = 53), respectively. (h) Reorientation angle histograms with peaks (± SDs) at 46 ± 28 degrees in SHU102 cells (left, n = 269), at 36 ± 13 degrees in HCB1336/pYS10 cells (middle, n = 285), and at 58 ± 29 degrees and 139 ± 30 degrees in HCB1336/pSHU61 cells (right, n = 444).