He Shichang, Sun Huijian, Chen Qing, Yang Yanlong, Zhou Zixin, Chang Saiwen, Lu Shuaiqi, Liang Zhencai, Yang Julan, Fei Xiao
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi, 830046, China.
Bayingolin Mongolian Autonomous Prefecture Agricultural Science Research Institute, Korla, 841000, China.
BMC Plant Biol. 2025 Jul 11;25(1):901. doi: 10.1186/s12870-025-06797-z.
The development of stomatal lineage cells in terrestrial plants is tightly regulated by epidermal patterning factors (EPFs/EPFLs) and their downstream receptors, including ERECTA, TOO MANY MOUTHS (TMM), and SOMATIC EMBRYOGENESIS RECEPTOR KINASEs (SERKs). These components form co-receptor complexes that activate the MAPK signaling cascade, playing critical roles in stomatal development, stress responses, and signal transduction. However, the EPF-ERECTA-TMM signaling network remains largely unexplored in cotton (Gossypium spp.). In this study, we performed a genome-wide identification and characterization of the EPF/EPFL, ERECTA, TMM, and SERK gene families in four cotton species (G. hirsutum, G. barbadense, G. arboreum, and G. raimondii), identifying 135 EPF/EPFL, 18 ERECTA, 6 TMM, and 90 SERK genes. Bioinformatics analyses-including gene collinearity, protein domain structure, cis-regulatory elements, and protein-protein interaction predictions-revealed functional divergence and stress-related regulatory potential across these families. Expression profiling in G. hirsutum indicated that several candidate genes, such as GhEPFL1-1, GhER1, and GhSERK17, are responsive to abiotic stresses. To validate these computational predictions, functional assays were conducted. Virus-induced gene silencing (VIGS) of GhEPFL1-1, GhER1, and GhSERK17 led to increased stomatal density and reduced drought tolerance, confirming their roles in stress adaptation. Furthermore, luciferase complementation imaging in Nicotiana benthamiana demonstrated direct interactions between GhEPFL1-1 and GhER1, and co-receptor complex formation with GhSERK17, consistent with molecular docking simulations. Collectively, this study lays a theoretical foundation for further exploration of the EPF/EPFL-mediated peptide-receptor signaling pathway in cotton and its potential application in breeding for stress resilience.
陆生植物气孔谱系细胞的发育受到表皮模式因子（EPFs/EPFLs）及其下游受体的严格调控，这些受体包括ERECTA、过多的口器（TMM）和体细胞胚胎发生受体激酶（SERKs）。这些成分形成共受体复合物，激活MAPK信号级联反应，在气孔发育、应激反应和信号转导中发挥关键作用。然而，EPF-ERECTA-TMM信号网络在棉花（棉属物种）中仍 largely未被探索。在本研究中，我们对四种棉花物种（陆地棉、海岛棉、亚洲棉和雷蒙德氏棉）中的EPF/EPFL、ERECTA、TMM和SERK基因家族进行了全基因组鉴定和表征，鉴定出135个EPF/EPFL、18个ERECTA、6个TMM和90个SERK基因。生物信息学分析——包括基因共线性、蛋白质结构域结构、顺式调控元件和蛋白质-蛋白质相互作用预测——揭示了这些家族之间的功能差异和与应激相关的调控潜力。陆地棉中的表达谱分析表明，一些候选基因，如GhEPFL1-1、GhER1和GhSERK17，对非生物胁迫有响应。为了验证这些计算预测，进行了功能测定。GhEPFL1-1、GhER1和GhSERK17的病毒诱导基因沉默（VIGS）导致气孔密度增加和耐旱性降低，证实了它们在应激适应中的作用。此外，烟草中的荧光素酶互补成像证明了GhEPFL1-1和GhER1之间的直接相互作用，以及与GhSERK17形成共受体复合物，这与分子对接模拟一致。总的来说，本研究为进一步探索棉花中EPF/EPFL介导的肽-受体信号通路及其在抗逆育种中的潜在应用奠定了理论基础。
