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. 2017 Jun 5;8:15707. doi: 10.1038/ncomms15707

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Copyright © 2017, The Author(s)

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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(a) Ancestral cells (strain Y-1140) typically occur as dyads or single cells, with occasional clusters of <8 cells. (b) All populations quickly evolved multicellular (‘snowflake') strains, which occur almost exclusively as large clusters (Supplementary Data 1). (c) Derived unicells resemble the ancestral form and were present in all ten populations at the end of the experiment. (d) Visual comparison of settling in 3 ml overnight cultures after 10 m in ancestral, unicellular and snowflake isolates. (e) Growth parameters of ancestral (open circle; n=1), derived snowflake (n=10) and derived unicellular (n=10) lineages. Points show mean parameter values estimated by fitting four parameter Gompertz models to each of four replicate populations per isolate. Both inflection point (t(18.0)=−11.13, P<0.0001 and maximum growth rate are significantly different between derived snowflakes and unicells (t(18.0)=5.72, P<0.0001; Supplementary Data 2); shaded ellipses show 95% CI. (f) Calcofluor white fluorescence of a group of three snowflakes. (g) Cell attachment within snowflakes occurs at bud scars (areas of heightened fluorescence, indicated by white arrows), reflecting continued association of daughter cells following division. All scale bars, 10 μm.