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. 2021 May 25;11:10894. doi: 10.1038/s41598-021-90225-3

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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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d-rR pairs spawn earlier than Kiyosu pairs, probably because d-rR male is more motivative to mating than Kiyosu male. (a) Time-course data of 30-min analyses of sexual behavior are shown as raster plots. Yellow, black, cyan and red bands represent the timing and duration of following, courtship, clasping and spawning, respectively. Red triangles are put on the right of each red bands for easy detection of them. (b,c) The latency data were further analyzed using Kaplan–Meier plots. Yellow and Blue curves represent Kiyosu and d-rR, respectively. d-rR pairs showed first following and spawning earlier than Kiyosu pairs (^†P < 0.1, **P < 0.001, by Log-Rank tests). (d–g) Various parameters were measured and compared between Kiyosu and d-rR. Yellow and Blue dots represent Kiyosu and d-rR, respectively. The frequency of courtship before spawning was significantly higher in d-rR than Kiyosu (e), while no significant difference was detected in the percentage of the time spent for following before spawning (d), the percentage of following with successful courtship before spawning (f), or the number of clasping ceased by female before spawning (g) (*P < 0.05, by Mann–Whitney U tests).