Cui Fengshuo, Wang Kang, Qi Haoran, Shen Tengfei, Chen Caihui, Zhong Yongda, Xu Meng
State Key Laboratory of Tree Genetics and Breeding, Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China.
Jiangsu Key Laboratory for Conservation and Utilization of Plant Resources, Institute of Botany, Jiangsu Province and Chinese Academy of Sciences (Nanjing Botanical Garden Mem. Sun Yat-Sen), Nanjing 210014, China.
Int J Mol Sci. 2025 Aug 23;26(17):8204. doi: 10.3390/ijms26178204.
The WUSCHEL-related homeobox (WOX) gene family is integral to plant growth and development. Here, we identified 14 genes from the genome and analyzed their phylogeny, conserved features, and expression patterns. Phylogenetic inference grouped CcWOX into the Ancient, Intermediate, and WUS clades, consistent with other plant lineages. Expression profiling across seven tissues/organs, together with qRT-PCR validation, revealed tissue-biased expression for several members (e.g., floral or root enrichment), suggesting gene-specific roles during development. Using AlphaFold3, we predicted monomeric structures for CcWOX proteins and an interface model compatible with an interaction between CcWOX3 and CcLBD33. Consistently, bimolecular fluorescence complementation (BiFC) in detected nuclear YFP signals for cEYFP-CcWOX3 + nEYFP-CcLBD33 relative to appropriate negative controls, confirming a physical interaction in plant cells. While these findings support a putative WOX-LBD interaction module in , the regulatory functions remain to be established. Overall, this work provides a framework for dissecting the CcWOX family in and illustrates how AI-assisted structure prediction can be integrated with cell-based assays to accelerate hypothesis generation in plant developmental biology.
WUSCHEL相关同源异型框(WOX)基因家族对植物生长发育至关重要。在此,我们从基因组中鉴定出14个基因,并分析了它们的系统发育、保守特征和表达模式。系统发育推断将CcWOX分为古老、中间和WUS分支,这与其他植物谱系一致。对七个组织/器官进行的表达谱分析以及qRT-PCR验证揭示了几个成员的组织偏向性表达(例如,在花或根中富集),表明这些基因在发育过程中具有特定作用。利用AlphaFold3,我们预测了CcWOX蛋白的单体结构以及与CcWOX3和CcLBD33之间相互作用兼容的界面模型。同样,在烟草中进行的双分子荧光互补(BiFC)检测到相对于适当阴性对照,cEYFP-CcWOX3 + nEYFP-CcLBD33有核YFP信号,证实了植物细胞中的物理相互作用。虽然这些发现支持烟草中存在一个假定的WOX-LBD相互作用模块,但其调控功能仍有待确定。总体而言,这项工作为剖析烟草中的CcWOX家族提供了一个框架,并说明了如何将人工智能辅助的结构预测与基于细胞的分析相结合,以加速植物发育生物学中假设的产生。