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However, the molecular mechanisms of flavonoid metabolism mediated by MYB repressors in B. napus are still not well understood. In this study, we identify an R2R3-MYB transcription factor, BnaC03.MYB56, which regulates flavonoid metabolism and oil content in B. napus. Overexpression of BnaC03.MYB56 reduced flavonoid metabolite levels and significantly increased seed oil content by 11.0% to11.4% compared to the wild type, whereas the mutant lines exhibited the opposite phenotype. BnaC03.MYB56 directly binds to the TAACGA motif in the promoters of BnaC08.TT6, which catalyzes the synthesis of dihydrokaempferol, and represses its expression. In addition, BnaA09.MYB61 interacts with BnaC03.MYB56 and antagonizes its transcriptional repression of BnaC08.TT6. BnaA09.MYB61 positively regulates flavonoid metabolism and oil content through activating the expression of BnaC08.TT6. Collectively, this study develops an antagonistic BnaC03.MYB56-BnaA09.MYB61 module that fine-tunes flavonoid metabolism to modulate seed oil content, providing a new target for improving oil content in B. napus breeding.<\/jats:p>","DOI":"10.1093\/plphys\/kiag677","type":"journal-article","created":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T11:47:42Z","timestamp":1789040862000},"source":"Crossref","is-referenced-by-count":0,"title":["BnaC03.MYB56 and BnaA09.MYB61 antagonistically regulate flavonoid metabolism in\n                    <i>Brassica napus<\/i>"],"prefix":"10.1093","volume":"202","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-8876-7171","authenticated-orcid":false,"given":"Youli","family":"Ao","sequence":"first","affiliation":[{"name":"Integrative Science Center of Germplasm Creation in Western China (CHONGQING) Science City and Southwest University, College of Agronomy and Biotechnology, Southwest University , Beibei, Chongqing 400715 ,","place":["China"]},{"name":"Academy of Agricultural Sciences, Southwest University , Beibei, Chongqing 400715 ,","place":["China"]},{"name":"Engineering Research Center of South Upland Agriculture, Ministry of Education , Chongqing 400715 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-1377-3452","authenticated-orcid":false,"given":"Jinxiong","family":"Xu","sequence":"additional","affiliation":[{"name":"Integrative Science Center of Germplasm Creation in Western China (CHONGQING) Science City and Southwest University, College of Agronomy and Biotechnology, Southwest University , Beibei, Chongqing 400715 ,","place":["China"]},{"name":"Academy of Agricultural Sciences, Southwest University , Beibei, Chongqing 400715 ,","place":["China"]},{"name":"Engineering Research Center of South Upland Agriculture, Ministry of Education , Chongqing 400715 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