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<p>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 (<i>Gossypium</i> spp.). In this study, we performed a genome-wide identification and characterization of the <i>EPF/EPFL</i>, <i>ERECTA</i>, <i>TMM</i>, and <i>SERK</i> gene families in four cotton species (<i>G. hirsutum</i>, <i>G. barbadense</i>, <i>G. arboreum</i>, and <i>G. raimondii</i>), identifying 135 <i>EPF/EPFL</i>, 18 <i>ERECTA</i>, 6 <i>TMM</i>, and 90 <i>SERK</i> 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 <i>G. hirsutum</i> indicated that several candidate genes, such as <i>GhEPFL1-1</i>, <i>GhER1</i>, and <i>GhSERK17</i>, are responsive to abiotic stresses. To validate these computational predictions, functional assays were conducted. Virus-induced gene silencing (VIGS) of <i>GhEPFL1-1</i>, <i>GhER1</i>, and <i>GhSERK17</i> led to increased stomatal density and reduced drought tolerance, confirming their roles in stress adaptation. Furthermore, luciferase complementation imaging in <i>Nicotiana benthamiana</i> demonstrated direct interactions between <i>GhEPFL1-1</i> and <i>GhER1</i>, and co-receptor complex formation with <i>GhSERK17</i>, 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.</p>

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Comprehensive identification of cotton EPF/EPFL receptors and functional characterization of the GhEPFL1-1-GhER1 module in drought tolerance

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年7月11日

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  • Shichang He,
  • Huijian Sun,
  • Qing Chen,
  • Yanlong Yang,
  • Zixin Zhou,
  • Saiwen Chang,
  • Shuaiqi Lu,
  • Zhencai Liang,
  • Julan Yang,
  • Xiao fei

摘要

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.