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

Abstract 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 co...

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Main Authors: Shichang He, Huijian Sun, Qing Chen, Yanlong Yang, Zixin Zhou, Saiwen Chang, Shuaiqi Lu, Zhencai Liang, Julan Yang, Xiao fei
Format: Article
Language:English
Published: BMC 2025-07-01
Series:BMC Plant Biology
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Online Access:https://doi.org/10.1186/s12870-025-06797-z
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author Shichang He
Huijian Sun
Qing Chen
Yanlong Yang
Zixin Zhou
Saiwen Chang
Shuaiqi Lu
Zhencai Liang
Julan Yang
Xiao fei
author_facet Shichang He
Huijian Sun
Qing Chen
Yanlong Yang
Zixin Zhou
Saiwen Chang
Shuaiqi Lu
Zhencai Liang
Julan Yang
Xiao fei
author_sort Shichang He
collection DOAJ
description Abstract 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.
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spelling doaj-art-d0662470ee9b4d13a6e3fca02df3e9df2025-08-20T03:04:27ZengBMCBMC Plant Biology1471-22292025-07-0125112210.1186/s12870-025-06797-zComprehensive identification of cotton EPF/EPFL receptors and functional characterization of the GhEPFL1-1-GhER1 module in drought toleranceShichang He0Huijian Sun1Qing Chen2Yanlong Yang3Zixin Zhou4Saiwen Chang5Shuaiqi Lu6Zhencai Liang7Julan Yang8Xiao fei9Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang UniversityBayingolin Mongolian Autonomous Prefecture Agricultural Science Research InstituteXinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang UniversityResearch Institute of Economic Crops, Xinjiang Academy of Agricultural SciencesResearch Institute of Economic Crops, Xinjiang Academy of Agricultural SciencesXinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang UniversityXinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang UniversityXinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang UniversityXinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang UniversityXinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang UniversityAbstract 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.https://doi.org/10.1186/s12870-025-06797-zEPF/EPFLGene family analysisCottonDrought stressGhEPFL1-1GhER1
spellingShingle Shichang He
Huijian Sun
Qing Chen
Yanlong Yang
Zixin Zhou
Saiwen Chang
Shuaiqi Lu
Zhencai Liang
Julan Yang
Xiao fei
Comprehensive identification of cotton EPF/EPFL receptors and functional characterization of the GhEPFL1-1-GhER1 module in drought tolerance
BMC Plant Biology
EPF/EPFL
Gene family analysis
Cotton
Drought stress
GhEPFL1-1
GhER1
title Comprehensive identification of cotton EPF/EPFL receptors and functional characterization of the GhEPFL1-1-GhER1 module in drought tolerance
title_full Comprehensive identification of cotton EPF/EPFL receptors and functional characterization of the GhEPFL1-1-GhER1 module in drought tolerance
title_fullStr Comprehensive identification of cotton EPF/EPFL receptors and functional characterization of the GhEPFL1-1-GhER1 module in drought tolerance
title_full_unstemmed Comprehensive identification of cotton EPF/EPFL receptors and functional characterization of the GhEPFL1-1-GhER1 module in drought tolerance
title_short Comprehensive identification of cotton EPF/EPFL receptors and functional characterization of the GhEPFL1-1-GhER1 module in drought tolerance
title_sort comprehensive identification of cotton epf epfl receptors and functional characterization of the ghepfl1 1 gher1 module in drought tolerance
topic EPF/EPFL
Gene family analysis
Cotton
Drought stress
GhEPFL1-1
GhER1
url https://doi.org/10.1186/s12870-025-06797-z
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