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However, whether CI consistently becomes associated or coupled with other host\u2010related forms of reproductive isolation (RI) to impede the transfer of endosymbionts between hybridizing populations and further the divergence process remains an open question. Here, we show that varying degrees of pre\u2010 and postmating RI exist among allopatric populations of two interbreeding cherry\u2010infesting tephritid fruit flies (<jats:italic>Rhagoletis cingulata<\/jats:italic>and<jats:italic>R<\/jats:italic>.<jats:italic>indifferens<\/jats:italic>) across North America. These flies display allochronic and sexual isolation among populations, as well as unidirectional reductions in egg hatch in hybrid crosses involving southwestern USA males. All populations are infected by a<jats:italic>Wolbachia<\/jats:italic>strain,<jats:italic>w<\/jats:italic>Cin2, whereas a second strain,<jats:italic>w<\/jats:italic>Cin3, only co\u2010infects flies from the southwest USA and Mexico. Strain<jats:italic>w<\/jats:italic>Cin3 is associated with a unique mitochondrial DNA haplotype and unidirectional postmating RI, implicating the strain as the cause of CI. When coupled with nonendosymbiont RI barriers, we estimate the strength of CI associated with<jats:italic>w<\/jats:italic>Cin3 would not prevent the strain from introgressing from infected southwestern to uninfected populations elsewhere in the USA if populations were to come into secondary contact and hybridize. In contrast, cytoplasmic\u2013nuclear coupling may impede the transfer of<jats:italic>w<\/jats:italic>Cin3 if Mexican and USA populations were to come into contact. We discuss our results in the context of the general paucity of examples demonstrating stable<jats:italic>Wolbachia<\/jats:italic>hybrid zones and whether the spread of<jats:italic>Wolbachia<\/jats:italic>among taxa can be constrained in natural hybrid zones long enough for the endosymbiont to participate in speciation.<\/jats:p>","DOI":"10.1111\/mec.16157","type":"journal-article","created":{"date-parts":[[2021,8,29]],"date-time":"2021-08-29T10:36:55Z","timestamp":1630233415000},"page":"2935-2950","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Testing the potential contribution of<i>Wolbachia<\/i>to speciation when cytoplasmic incompatibility becomes associated with host\u2010related reproductive isolation"],"prefix":"10.1111","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3115-6599","authenticated-orcid":false,"given":"Daniel J.","family":"Bruzzese","sequence":"first","affiliation":[{"name":"Department of Biological Sciences University of Notre Dame Notre Dame IN USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8307-9831","authenticated-orcid":false,"given":"Hannes","family":"Schuler","sequence":"additional","affiliation":[{"name":"Faculty of Science and Technology Free University of Bozen\u2010Bolzano Bozen\u2010Bolzano Italy"},{"name":"Competence Centre for Plant Health Free University of Bozen\u2010Bolzano Bozen\u2010Bolzano Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1509-0147","authenticated-orcid":false,"given":"Thomas M.","family":"Wolfe","sequence":"additional","affiliation":[{"name":"Department of Forest and Soil Sciences Boku, University of Natural Resources and Life Sciences Vienna Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mary M.","family":"Glover","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences University of Notre Dame Notre Dame IN USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph V.","family":"Mastroni","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences University of Notre Dame Notre Dame IN USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0903-6590","authenticated-orcid":false,"given":"Meredith M.","family":"Doellman","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences University of Notre Dame Notre Dame IN USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1851-8911","authenticated-orcid":false,"given":"Cheyenne","family":"Tait","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences University of Notre Dame Notre Dame IN USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wee L.","family":"Yee","sequence":"additional","affiliation":[{"name":"United States Department of Agriculture Temperate Tree Fruit &amp; Vegetable Research Unit Agricultural Research Service Wapato WA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Rull","sequence":"additional","affiliation":[{"name":"Instituto de Ecolog\u00eda A.C. Xalapa M\u00e9xico"},{"name":"LIEMEN\u2010Divisi\u00f3n Control Biol\u00f3gico de Plagas PROIMI Biotecnolog\u00eda\u2010CONICET Tucum\u00e1n Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Aluja","sequence":"additional","affiliation":[{"name":"Instituto de Ecolog\u00eda A.C. 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