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Cyanogenesis, the ability to produce hydrogen cyanide (<jats:styled-content style=\"fixed-case\">HCN<\/jats:styled-content>) after tissue damage, occurs in &gt;3000 plant species and exists as a discrete polymorphism in white clover. This polymorphism is spatially distributed in recurrent clines, with higher frequencies of cyanogenic plants in warmer climates. The <jats:styled-content style=\"fixed-case\">HCN<\/jats:styled-content> autotoxicity hypothesis proposes that cyanogenic plants are selected against where frosts are common, as freezing liberates <jats:styled-content style=\"fixed-case\">HCN<\/jats:styled-content> and could impair cellular respiration.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We tested the <jats:styled-content style=\"fixed-case\">HCN<\/jats:styled-content> autotoxicity hypothesis using a freezing chamber to examine survival, tissue damage, and physiological recovery as assessed via chlorophyll fluorescence following mild and severe freezing treatments. We utilized 65 genotypes from a single polymorphic population to eliminate effects of population structure.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Results<\/jats:title><jats:p>Cyanogenic plants did not differ from acyanogenic plants in survival, tissue damage, or recovery following freezing. However, plants producing either of the two required cyanogenic precursors had lower survival and tissue damage after freezing than plants lacking both precursors.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>These results suggest that freezing\u2010induced <jats:styled-content style=\"fixed-case\">HCN<\/jats:styled-content> toxicity is unlikely to be responsible for the maintenance of the cyanogenesis polymorphism in white clover. However, energetic trade\u2010offs associated with costs of producing the cyanogenic precursors may confer a fitness benefit to acyanogenic plants under stressful climatic conditions. The lack of evidence for <jats:styled-content style=\"fixed-case\">HCN<\/jats:styled-content> toxicity suggests that cyanogenic clover uses physiological mechanisms mediated by \u03b2\u2010cyanoalanine synthase and alternative oxidase to maintain cellular function in the presence of <jats:styled-content style=\"fixed-case\">HCN<\/jats:styled-content>.<\/jats:p><\/jats:sec>","DOI":"10.1002\/ajb2.1134","type":"journal-article","created":{"date-parts":[[2018,8,7]],"date-time":"2018-08-07T06:52:06Z","timestamp":1533624726000},"page":"1224-1231","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":29,"title":["Freeze\u2010induced cyanide toxicity does not maintain the cyanogenesis polymorphism in white clover (<i>Trifolium repens<\/i>)"],"prefix":"10.1002","volume":"105","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3398-7377","authenticated-orcid":false,"given":"Nicholas J.","family":"Kooyers","sequence":"first","affiliation":[{"name":"Department of Biology University of Louisiana  Lafayette LA 70504 USA"},{"name":"Department of Integrative Biology University of South Florida  Tampa FL 33620 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bradley","family":"Hartman Bakken","sequence":"additional","affiliation":[{"name":"Division of Biology Kansas State University  Manhattan KS 66506 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark C.","family":"Ungerer","sequence":"additional","affiliation":[{"name":"Division of Biology Kansas State University  Manhattan KS 66506 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8338-3638","authenticated-orcid":false,"given":"Kenneth M.","family":"Olsen","sequence":"additional","affiliation":[{"name":"Department of Biology Washington University  St. Louis MO 63130 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,8,6]]},"reference":[{"key":"e_1_2_6_2_1","volume-title":"Ethylene in plant biology","author":"Abeles F. 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