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. 2017 Dec 14;7:17550. doi: 10.1038/s41598-017-18019-0

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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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Natural variation of social behaviours in P. pacificus independent of Ppa-npr-1. (a) Clumping/bordering assay for the P. pacificus reference strain RS2333, which shows a solitary behaviour under laboratory conditions, characterized by low levels of both clumping and bordering behaviours. (b) Clumping/bordering assay for the P. pacificus RSB001 strain, isolated at 2327 m.a.s.l. on La Réunion Island, which shows strong clumping and bordering behaviours under laboratory conditions. (c) Map of La Réunion Island showing the locations from which P. pacificus nematode strains have been isolated. Social strains, belonging to the phylogenetic lineage B, have been isolated from only two locations above 2000 m.a.s.l. (modified from OpenStreetMap^® under the Open Database License (ODbL), © OpenStreetMap contributors (https://www.openstreetmap.org/copyright)). (d) Regulation of bordering behaviour by oxygen levels: comparison of the dynamic changes in bordering behaviour between RSB001 and RS2333 strains during oxygen shifts from 21% to 16%, with a 1% decrement occurring every 30 min (modified from Moreno et al.¹⁵). Black dots represent the mean and thin lines represent the standard error of the mean (SEM). (e) Ω-turn rate response of RS2333 to 21% −>20% −>21% oxygen shifts on a lawn of Escherichia coli OP50¹⁵. The black line represents the mean Ω-turn and the grey area, the SEM. (f) Bordering behaviour of RS2333, RSB001 and the Ppa-npr-1 mutants generated using CRISPR/Cas9 system (Suppl. Table S1). In all bar-plots arrows represent the standard error of the mean (SEM). For statistical analysis see Suppl. Table S2.