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We identified 26 SPL8\u2010interacting transcription factors by yeast two\u2010hybrid screening and bimolecular fluorescence complementation (BiFC) assays, with YABBY (YAB) and TEOSINTE BRANCHED 1\/CYCLOIDEA\/PCF (TCP) family emerging as the predominant interactors. Among these, YAB3 and TCP19 were selected for detailed characterization. Their interactions with SPL8, as well as with each other, were confirmed in plant nuclei by fluorescence resonance energy transfer\u2013fluorescence lifetime imaging microscopy (FRET\u2010FLIM). Chromatin immunoprecipitation sequencing (ChIP\u2010seq), ChIP\u2010qPCR, yeast one\u2010hybrid, and dual\u2010luciferase assays demonstrated that SPL8 directly binds the regulatory regions of\n                    <jats:italic>YAB3<\/jats:italic>\n                    and\n                    <jats:italic>TCP19<\/jats:italic>\n                    and activates their transcription. Dexamethasone\u2010inducible expression assays further showed that this induction occurs independently of de novo protein synthesis. Expression analyses revealed overlapping spatial and temporal patterns of\n                    <jats:italic>SPL8<\/jats:italic>\n                    ,\n                    <jats:italic>YAB3<\/jats:italic>\n                    , and\n                    <jats:italic>TCP19<\/jats:italic>\n                    during early anther development, particularly in tapetal cells, microspore mother cells, and vascular tissues. Genetic and cellular analyses of promoter\u2010driven SRDX repression lines showed that\n                    <jats:italic>SPL8<\/jats:italic>\n                    ,\n                    <jats:italic>YAB3<\/jats:italic>\n                    , and\n                    <jats:italic>TCP19<\/jats:italic>\n                    regulate anther morphogenesis, tapetal differentiation, meiotic progression, and microspore development. Although individual family members exhibited functional redundancy, their repression caused severe developmental defects and male sterility. Together, these findings establish the\n                    <jats:italic>SPL8<\/jats:italic>\n                    \u2013\n                    <jats:italic>YAB3<\/jats:italic>\n                    \/\n                    <jats:italic>TCP19<\/jats:italic>\n                    module as a multilayered regulatory system in which\n                    <jats:italic>SPL8<\/jats:italic>\n                    controls anther development through both transcriptional activation of\n                    <jats:italic>YAB3<\/jats:italic>\n                    and\n                    <jats:italic>TCP19<\/jats:italic>\n                    and physical interaction with these proteins, expanding the regulatory framework of\n                    <jats:italic>SPL<\/jats:italic>\n                    \u2010mediated reproductive development.\n                  <\/jats:p>","DOI":"10.1111\/tpj.71160","type":"journal-article","created":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T04:59:37Z","timestamp":1791781177000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The\n                    <i>SPL8\u2010YAB3\/TCP19<\/i>\n                    module integrates transcriptional and protein\u2013protein regulatory mechanisms to orchestrate anther development in Arabidopsis"],"prefix":"10.1111","volume":"128","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-8822-8407","authenticated-orcid":false,"given":"Tingting","family":"Li","sequence":"first","affiliation":[{"name":"School of Life Science Shanxi University  Taiyuan 030006 China"},{"name":"Institute of Applied Biology Shanxi University  Taiyuan 030006 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