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Although reproductive isolation is inherently a composite process, most empirical studies have focused on individual barriers in isolation, limiting our ability to capture their joint effects, particularly in systems undergoing evolutionary transitions such as shifts in mating system. Here, we provide a comprehensive, life-cycle-wide quantification of reproductive isolation between two closely related species of the Erysimum incanum complex that differ markedly in mating system: the predominantly selfing E. incanum and the outcrossing E. wilczekianum. By integrating ecological, phenological, behavioural, and post-pollination components, we show that total reproductive isolation is nearly complete (T = 0.999) but largely driven by pre-pollination barriers. Ecogeographical differentiation and, most prominently, pollinator-mediated isolation dominates, with pollinators showing a strong bias towards E. wilczekianum. Floral traits associated with mating system divergence, particularly flower size, emerge as key drivers of assortative mating, supporting their role as \u2018magic traits\u2019 linking ecological divergence and reproductive isolation. In contrast, post-pollination barriers are weaker but strongly asymmetric. Hybrid seed formation is largely prevented when E. wilczekianum acts as the maternal parent, whereas reciprocal crosses are relatively successful. Despite reduced germination, hybrids show enhanced growth and no evidence of hybrid breakdown, suggesting that intrinsic incompatibilities remain incomplete. Overall, our results indicate that mating system divergence reshapes the entire architecture of reproductive isolation, highlighting the importance of integrative approaches to understand how multiple barriers interact during early stages of speciation.<\/jats:p>","DOI":"10.1093\/aobpla\/plag052","type":"journal-article","created":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T11:37:46Z","timestamp":1791027466000},"source":"Crossref","is-referenced-by-count":0,"title":["Reproductive isolation emerges from coordinated barriers during mating system divergence in plants"],"prefix":"10.1093","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5421-1100","authenticated-orcid":false,"given":"Ana","family":"Garc\u00eda-Mu\u00f1oz","sequence":"first","affiliation":[{"name":"BioChange Network, Departamento de Gen\u00e9tica, Universidad de Granada , Facultad de Ciencias. Avenida de Fuente Nueva s\/n, Granada 18071 ,","place":["Spain"]},{"name":"Departamento de Biolog\u00eda Vegetal y Ecolog\u00eda, Universidad de Sevilla , Avenida de la Reina Mercedes 6, Sevilla 41012 ,","place":["Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5066-7247","authenticated-orcid":false,"given":"Camilo","family":"Ferr\u00f3n","sequence":"additional","affiliation":[{"name":"BioChange Network, Departamento de Gen\u00e9tica, Universidad de Granada , Facultad de Ciencias. Avenida de Fuente Nueva s\/n, Granada 18071 ,","place":["Spain"]},{"name":"\u00c1rea de Biodiversidad y Conservaci\u00f3n, Departamento de Biolog\u00eda y Geolog\u00eda, F\u00edsica y Qu\u00edmica Inorg\u00e1nica, Universidad Rey Juan Carlos , Calle Tulip\u00e1n s\/n, M\u00f3stoles, Madrid 28931,\u00a0","place":["Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8194-7751","authenticated-orcid":false,"given":"Enrica","family":"Olivieri","sequence":"additional","affiliation":[{"name":"BioChange Network, Departamento de Gen\u00e9tica, Universidad de Granada , Facultad de Ciencias. Avenida de Fuente Nueva s\/n, Granada 18071 ,","place":["Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-0849-4759","authenticated-orcid":false,"given":"Esther","family":"Funes-Ligero","sequence":"additional","affiliation":[{"name":"BioChange Network, Departamento de Gen\u00e9tica, Universidad de Granada , Facultad de Ciencias. Avenida de Fuente Nueva s\/n, Granada 18071 ,","place":["Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0533-6817","authenticated-orcid":false,"given":"Mohamed","family":"Abdelaziz","sequence":"additional","affiliation":[{"name":"BioChange Network, Departamento de Gen\u00e9tica, Universidad de Granada , Facultad de Ciencias. 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