Frontiers in plant science · 2026
The ADF4-RCAR12 module regulates ABA-mediated drought tolerance in Arabidopsis.
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Li Xiaoyi, Feng Yihan, He Jiahan, Li Yufei, Li Xufeng, Wang Jianmei, Yang Yi, Wang Maohua
Аннотация
INTRODUCTION: Drought is one of the most severe abiotic stresses limiting crop productivity, and abscisic acid (ABA)-induced stomatal closure is a key mechanism underlying plant drought resistance. Although ABA signaling has been extensively characterized in Arabidopsis thaliana, the role and regulatory mechanisms of microfilaments (F-actin) in this process remain poorly understood. Previously, we identified the actin-binding protein CAP1 as an interactor of the ABA receptor RCAR12 and a phosphorylation target of OPEN STOMATA 1 (OST1), suggesting a potential link between actin dynamics and ABA signaling.
METHODS: To investigate the role of Actin Depolymerizing Factor 4 (ADF4) in ABA signaling, we examined its interaction with RCAR12 using bimolecular fluorescence complementation (BiFC), GST pull-down, and co-immunoprecipitation (Co-IP) assays. Genetic interactions between ADF4 and RCAR12 were further analyzed using ADF4-overexpressing lines in the ABA receptor quadruple mutant 1124 (pyr1 pyl1 pyl2 pyl4) and the adf4 × OE-RCAR12 genotype. ABA responses and drought tolerance were evaluated through seed germination, cotyledon greening, stomatal closure, and drought assays.
RESULTS: ADF4 physically interacted with RCAR12 in vivo and in vitro. Overexpression of ADF4 only weakly rescued the ABA-insensitive phenotype of the 1124 quadruple mutant during seed germination and seedling establishment, but completely restored its drought sensitivity. In addition, the adf4 × OE-RCAR12 genotype partially rescued the defects of the adf4 mutant in ABA-mediated seed germination, cotyledon greening, ABA-induced stomatal closure, and drought tolerance.
DISCUSSION: These findings demonstrate that ADF4 functionally interacts with the ABA receptor RCAR12 and that the two proteins act synergistically in regulating ABA responses. Our results establish a link between actin dynamics and ABA receptor-mediated signaling and suggest that ADF4-RCAR12 coordination contributes to ABA-induced stomatal regulation and drought tolerance in Arabidopsis.
METHODS: To investigate the role of Actin Depolymerizing Factor 4 (ADF4) in ABA signaling, we examined its interaction with RCAR12 using bimolecular fluorescence complementation (BiFC), GST pull-down, and co-immunoprecipitation (Co-IP) assays. Genetic interactions between ADF4 and RCAR12 were further analyzed using ADF4-overexpressing lines in the ABA receptor quadruple mutant 1124 (pyr1 pyl1 pyl2 pyl4) and the adf4 × OE-RCAR12 genotype. ABA responses and drought tolerance were evaluated through seed germination, cotyledon greening, stomatal closure, and drought assays.
RESULTS: ADF4 physically interacted with RCAR12 in vivo and in vitro. Overexpression of ADF4 only weakly rescued the ABA-insensitive phenotype of the 1124 quadruple mutant during seed germination and seedling establishment, but completely restored its drought sensitivity. In addition, the adf4 × OE-RCAR12 genotype partially rescued the defects of the adf4 mutant in ABA-mediated seed germination, cotyledon greening, ABA-induced stomatal closure, and drought tolerance.
DISCUSSION: These findings demonstrate that ADF4 functionally interacts with the ABA receptor RCAR12 and that the two proteins act synergistically in regulating ABA responses. Our results establish a link between actin dynamics and ABA receptor-mediated signaling and suggest that ADF4-RCAR12 coordination contributes to ABA-induced stomatal regulation and drought tolerance in Arabidopsis.