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Here, we examine the environmental factors that drive the evolution of parallel urban-rural clines in a Mendelian trait\u2014the cyanogenic antiherbivore defense of white clover (Trifolium repens). Previous work suggested urban-rural gradients in frost and snow depth could drive the evolution of reduced hydrogen cyanide (HCN) frequencies in urban populations. Here, we sampled over 700 urban and rural clover populations across 16 cities along a latitudinal transect in eastern North America. In each population, we quantified changes in the frequency of genotypes that produce HCN, and in a subset of the cities we estimated the frequency of the alleles at the two genes (CYP79D15 and Li) that epistatically interact to produce HCN. We then tested the hypothesis that cold climatic conditions are necessary for the evolution of cyanogenesis clines by comparing the strength of clines among cities located along a latitudinal gradient of winter temperature and frost exposure. Overall, half of the cities exhibited urban-rural clines in the frequency of HCN, whereby urban populations evolved lower HCN frequencies. Clines did not evolve in cities with the lowest temperatures and greatest snowfall, supporting the hypothesis that snow buffers plants against winter frost and constrains the formation of clines. By contrast, the strongest clines occurred in the warmest cities where snow and frost are rare, suggesting that alternative selective agents are maintaining clines in warmer cities. Some clines were driven by evolution at only CYP79D15, consistent with stronger and more consistent selection on this locus than on Li. Together, our results demonstrate that urban environments often select for similar phenotypes, but different selective agents and targets underlie the evolutionary response in different cities.<\/jats:p>","DOI":"10.1002\/evl3.163","type":"journal-article","created":{"date-parts":[[2020,3,25]],"date-time":"2020-03-25T05:46:45Z","timestamp":1585115205000},"page":"212-225","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Predicting the strength of urban-rural clines in a Mendelian polymorphism along a latitudinal gradient"],"prefix":"10.1093","volume":"4","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5921-2548","authenticated-orcid":false,"given":"James S.","family":"Santangelo","sequence":"first","affiliation":[{"name":"Department of Biology University of Toronto Mississauga Mississauga ON L5L 1C6 Canada"},{"name":"Centre for Urban Environments University of Toronto Mississauga Mississauga ON L5L 1C6 Canada"},{"name":"Department of Ecology and Evolutionary Biology University of Toronto Toronto ON M5S 3B2 Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1487-3254","authenticated-orcid":false,"given":"Ken A.","family":"Thompson","sequence":"additional","affiliation":[{"name":"Department of Zoology and Biodiversity Research Centre University of British Columbia Vancouver BC V6T 1Z4 Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beata","family":"Cohan","sequence":"additional","affiliation":[{"name":"Department of Biology University of Toronto Mississauga Mississauga ON L5L 1C6 Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jibran","family":"Syed","sequence":"additional","affiliation":[{"name":"Department of Biology University of Toronto Mississauga Mississauga ON L5L 1C6 Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7313-8236","authenticated-orcid":false,"given":"Rob W.","family":"Ness","sequence":"additional","affiliation":[{"name":"Department of Biology University of Toronto Mississauga Mississauga ON L5L 1C6 Canada"},{"name":"Centre for Urban Environments University of Toronto Mississauga Mississauga ON L5L 1C6 Canada"},{"name":"Department of Ecology and Evolutionary Biology University of Toronto Toronto ON M5S 3B2 Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9719-0522","authenticated-orcid":false,"given":"Marc T. J.","family":"Johnson","sequence":"additional","affiliation":[{"name":"Department of Biology University of Toronto Mississauga Mississauga ON L5L 1C6 Canada"},{"name":"Centre for Urban Environments University of Toronto Mississauga Mississauga ON L5L 1C6 Canada"},{"name":"Department of Ecology and Evolutionary Biology University of Toronto Toronto ON M5S 3B2 Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2020,6,1]]},"reference":[{"key":"2023041206474596400_","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1126\/science.1225977","article-title":"Insect herbivores drive real-time ecological and evolutionary change in plant populations","volume":"338","author":"Agrawal","year":"2012","journal-title":"Science"},{"key":"2023041206474596400_","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1038\/hdy.1973.84","article-title":"Field observations on the cyanogenesis polymorphism in Trifolium repens","volume":"31","author":"Angseesing","year":"1973","journal-title":"Heredity"},{"key":"2023041206474596400_","volume-title":"MuMIn: multi-model inference","author":"Barto\u0144","year":"2016"},{"key":"2023041206474596400_","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1146\/annurev-ecolsys-110617-062240","article-title":"(Non)parallel evolution","volume":"49","author":"Bolnick","year":"2018","journal-title":"Annu. 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