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Here, we identified a novel single\u2010gene\u2010controlled recessive genic male sterility (GMS) mutant,\n                    <jats:italic>defective in filament elongation 1<\/jats:italic>\n                    (\n                    <jats:italic>def1<\/jats:italic>\n                    ), which exhibits non\u2010elongated filaments, unopened glumes, and non\u2010exserted anthers, yet undergoes normal microsporogenesis and produces viable pollen. Map\u2010based cloning revealed that\n                    <jats:italic>def1<\/jats:italic>\n                    is caused by a loss\u2010of\u2010function mutation in the\n                    <jats:italic>ZmKO1<\/jats:italic>\n                    gene. Previous studies have identified\n                    <jats:italic>ZmKO1<\/jats:italic>\n                    and\n                    <jats:italic>ZmKO2<\/jats:italic>\n                    as the two putative KO\u2010encoding genes in maize. Although heterologous expression evidence suggested that only ZmKO1 possesses KO activity, we demonstrate that both ZmKO1 and ZmKO2 are functional KOs for GA biosynthesis. The\n                    <jats:italic>def1<\/jats:italic>\n                    mutant or\n                    <jats:italic>ZmKO1<\/jats:italic>\n                    knockout lines exhibited reduced plant height during early seedling development but recovered rapidly to wild\u2010type levels. In contrast,\n                    <jats:italic>ZmKO2<\/jats:italic>\n                    knockout lines displayed severe dwarfism and pollen\u2010less male sterility. These contrasting phenotypes demonstrate a clear functional divergence between the two genes. Utilising marker\u2010assisted backcrossing and CRISPR\/Cas9 editing, we generated ZmKO1\u2010based GMS lines across diverse genetic backgrounds. These lines maintained stable male sterility and could be efficiently propagated via exogenous GA\n                    <jats:sub>3<\/jats:sub>\n                    application under different genetic backgrounds and environments, demonstrating robust \u201cdual\u2010use\u201d properties. Data indicated that this dual\u2010use trait has no significant effect on other agronomic traits of the male sterile line and its derived hybrid combinations. Based on this, we established a novel two\u2010line hybrid seed production system in maize, termed the Gibberellin\u2010switchable Dual\u2010use Line (GDL), and successfully applied it in the hybrid seed production of Chuandan 99.\n                  <\/jats:p>","DOI":"10.1111\/pbi.70608","type":"journal-article","created":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T12:28:56Z","timestamp":1773836936000},"page":"4361-4377","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Engineering a Gibberellin\u2010Switchable Dual\u2010Use Line Based on Ent\u2010Kaurene Oxidase Gene\n                    <scp>\n                      <i>ZmKO1<\/i>\n                    <\/scp>\n                    Enables Two\u2010Line Hybrid Seed Production in Maize"],"prefix":"10.1111","volume":"24","author":[{"given":"Chuan","family":"Li","sequence":"first","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siyu","family":"Wang","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Xiang","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tao","family":"Yu","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rujun","family":"Wei","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenqian","family":"Mei","sequence":"additional","affiliation":[{"name":"Syngenta Biotech China  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaowei","family":"Liu","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Yuan","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongyang","family":"Yi","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shufeng","family":"Zhou","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chao","family":"Xia","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moju","family":"Cao","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hai","family":"Lan","sequence":"additional","affiliation":[{"name":"Maize Research Institute Sichuan Agricultural University  Chengdu Sichuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,3,18]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb2546"},{"issue":"1","key":"e_1_2_10_3_1","first-page":"75","article-title":"Cloning and Characterization of the Maize An1 Gene","volume":"7","author":"Bensen R. 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