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. 2020 Jul 30;10:12830. doi: 10.1038/s41598-020-69767-5

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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 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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Morphology and geometric morphometrics of the stylet. Micrographs of the stylet of an individual (a) M. lignano, (b) F1 hybrid, and (c) M. janickei. The placement of 60 landmarks along the stylet of an individual (d) M. lignano, (e) F1 hybrid, and (f) M. janickei. Note that we placed four fixed landmarks (filled red circles), two on the stylet base and two on the stylet tip, and 28 equally spaced sliding semi-landmarks (empty red circles) along each curved side of the stylet. The numbers indicate the order in which the landmarks were placed (note that the seminal vesicles always are to the left of the stylet). Visualization of thin-plate splines of the stylet derived from relative warp score analysis. Each panel shows the visualization of the mean relative warp score 1 (RWS1) observed for individuals of (g) M. lignano, (h) the F1 hybrids and (i) M. janickei. Thus, in general M. lignano has a relatively straight stylet tip and M. janickei has a stylet tip that curves drastically, while the hybrids have intermediate curvature. The scale bar in (e) represents 20 µm, and is applicable to all photomicrographic images.