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BpCYCD3;1 regulates organ growth by activating the cell cycle module in white birch
Linan Yue1, Xinyu Wang1, Zhimin Zheng1
1State Key Laboratory of Tree Genetics and Breeding, College of Forestry, Northeast Forestry University, Harbin 150040, China; The Center for Basic Forestry Research, College of Forestry, Northeast Forestry University, Harbin 150040, China.
Abstract:
Cyclin D proteins are key regulators of cell cycle progression and plant growth. Here, we identified BpCYCD3;1, a CYCD3 family member in birch, and investigated its function. Phylogenetic and sequence analyses showed that BpCYCD3;1 is evolutionarily conserved and contains canonical cell cycle-related domains. Promoter and expression analyses suggested responsiveness to environmental and hormonal signals, with preferential expression in leaves. BpCYCD3;1 overexpression promoted leaf expansion and and biomass accumulation, whereas CRISPR/Cas9 knockout reduced vegetative growth. Cytological analyses revealed increased cell density and epidermal cell number in overexpression lines, indicating enhanced cell proliferation. At the molecular level, BpCYCD3;1 upregulated G1/S transition regulators and DNA replication-related genes. Yeast two-hybrid and docking analyses suggested interaction between BpCYCD3;1 and CDKA;1, as well as the interaction between RBR and E2F transcription factors, and binding of E2FA/E2FB to downstream promoters. Together, these results indicate that BpCYCD3;1 promotes vegetative growth by activating the CDKA-RBR-E2F regulatory module, providing a potential target for improving tree growth and biomass production.
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