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We have identified\n amhy<\/jats:italic>\n ,\n dmrt1<\/jats:italic>\n ,\n gsdf<\/jats:italic>\n as male and\n foxl2<\/jats:italic>\n ,\n foxl3<\/jats:italic>\n ,\n cyp19a1a<\/jats:italic>\n as female sex determination pathway genes in Nile tilapia. However, the relationship among these genes is largely unclear. Here, we found that the gonads of\n dmrt1<\/jats:italic>\n ;\n cyp19a1a<\/jats:italic>\n double mutants developed as ovaries or underdeveloped testes with no germ cells irrespective of their genetic sex. In addition, the gonads of\n dmrt1<\/jats:italic>\n ;\n cyp19a1a<\/jats:italic>\n ;\n cyp19a1b<\/jats:italic>\n triple mutants still developed as ovaries. The gonads of\n foxl3<\/jats:italic>\n ;\n cyp19a1a<\/jats:italic>\n double mutants developed as testes, while the gonads of\n dmrt1<\/jats:italic>\n ;\n cyp19a1a<\/jats:italic>\n ;\n foxl3<\/jats:italic>\n triple mutants eventually developed as ovaries. In contrast, the gonads of\n amhy<\/jats:italic>\n ;\n cyp19a1a<\/jats:italic>\n ,\n gsdf<\/jats:italic>\n ;\n cyp19a1a<\/jats:italic>\n ,\n amhy<\/jats:italic>\n ;\n foxl2<\/jats:italic>\n ,\n gsdf<\/jats:italic>\n ;\n foxl2<\/jats:italic>\n double and\n amhy<\/jats:italic>\n ;\n cyp19a1a<\/jats:italic>\n ;\n cyp19a1b<\/jats:italic>\n ,\n gsdf<\/jats:italic>\n ;\n cyp19a1a<\/jats:italic>\n ;\n cyp19a1b<\/jats:italic>\n triple mutants developed as testes with spermatogenesis via up-regulation of\n dmrt1<\/jats:italic>\n in both somatic and germ cells. The gonads of\n amhy<\/jats:italic>\n ;\n foxl3<\/jats:italic>\n and\n gsdf<\/jats:italic>\n ;\n foxl3<\/jats:italic>\n double mutants developed as ovaries but with germ cells in spermatogenesis due to up-regulation of\n dmrt1<\/jats:italic>\n . Taking the respective ovary and underdeveloped testis of\n dmrt1<\/jats:italic>\n ;\n foxl3<\/jats:italic>\n and\n dmrt1<\/jats:italic>\n ;\n foxl2<\/jats:italic>\n double mutants reported previously into consideration, we demonstrated that once\n dmrt1<\/jats:italic>\n mutated, the gonad could not be rescued to functional testis by mutating any female pathway gene. The sex reversal caused by mutation of male pathway genes other than\n dmrt1<\/jats:italic>\n , including its upstream\n amhy<\/jats:italic>\n and downstream\n gsdf<\/jats:italic>\n , could be rescued by mutating female pathway gene. Overall, our data suggested that\n dmrt1<\/jats:italic>\n is the only male pathway gene tested indispensable for sex determination and functional testis development in tilapia.\n <\/jats:p>","DOI":"10.1371\/journal.pgen.1011210","type":"journal-article","created":{"date-parts":[[2024,3,27]],"date-time":"2024-03-27T13:35:05Z","timestamp":1711546505000},"page":"e1011210","update-policy":"https:\/\/doi.org\/10.1371\/journal.pgen.corrections_policy","source":"Crossref","is-referenced-by-count":42,"title":["Dmrt1 is the only male pathway gene tested indispensable for sex determination and functional testis development in tilapia"],"prefix":"10.1371","volume":"20","author":[{"given":"Shuangshuang","family":"Qi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4397-6997","authenticated-orcid":true,"given":"Shengfei","family":"Dai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6656-4832","authenticated-orcid":true,"given":"Xueyan","family":"Wei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanyuan","family":"He","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7547-0133","authenticated-orcid":true,"given":"Thomas D.","family":"Kocher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deshou","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0880-2783","authenticated-orcid":true,"given":"Minghui","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,3,27]]},"reference":[{"issue":"11","key":"pgen.1011210.ref001","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1038\/nrg.2017.60","article-title":"Vertebrate sex determination: evolutionary plasticity of a fundamental switch","volume":"18","author":"B Capel","year":"2017","journal-title":"Nat Rev Genet"},{"issue":"3","key":"pgen.1011210.ref002","doi-asserted-by":"crossref","first-page":"1237","DOI":"10.1152\/physrev.00044.2019","article-title":"Sex determination, gonadal sex 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