The TaSnRK1-TabHLH489 module integrates brassinosteroid and sugar signaling to regulate grain length in bread wheat

Regulation of grain size is a crucial strategy for improving crop yield and is also a fundamental aspect of developmental biology. However, the underlying molecular mechanisms governing grain development in wheat remain largely unknown. In this study, we identified a wheat atypical basic helix-loop-helix (bHLH) transcription factor, TabHLH489, which is tightly associated with grain length through genome-wide association study and map-based cloning. Knockout of TabHLH489 and its homologous genes resulted in increased grain length and weight, whereas overexpression led to decreased grain length and weight. TaSnRK1α1, the α-catalytic subunit of plant energy sensor SnRK1, interacted with and phosphorylated TabHLH489 to induce its degradation, thereby promoting wheat grain development. Sugar treatment induced TaSnRK1α1 protein accumulation while reducing TabHLH489 protein levels. Moreover, brassinosteroid (BR) promotes grain development by decreasing TabHLH489 expression through the transcription factor BRASSINAZOLE RESISTANT1 (BZR1). Importantly, natural variations in the promoter region of TabHLH489 affect the TaBZR1 binding ability, thereby influencing TabHLH489 expression. Taken together, our findings reveal that the TaSnRK1α1-TabHLH489 regulatory module integrates BR and sugar signaling to regulate grain length, presenting potential targets for enhancing grain size in wheat. To explore the functional mechanism of TabHLH489 in the development of wheat grains, we collected 5-day-post-anthesis (5-DPA) grains from Fielder and OE-TabHLH489, respectively. The plants were cultivated in a greenhouse under a 16-hour light (22 °C)/8-hour dark (16 °C) cycle, maintaining approximately 50% relative humidity. Each sample consisted of grains collected from five individuals, mixed to form a single biological repeat.