Nitric Oxide Enhances Drought Tolerance in Gossypium hirsutum L. via S-Nitrosylation of the Plasma Membrane H+-ATPase Isoform GhHA2 and Antioxidant Defense Activation

(1) Background: Nitric oxide (NO) serves as a crucial signaling molecule in plant abiotic stress responses. Although its role in enhancing drought resistance in cotton has been recognized, the specific mechanisms underlying this physiological and molecular regulation remain largely unexplored. This study aims to elucidate the multi-layered mechanisms by which NO modulates drought resistance in cotton; (2) Methods: Cotton seedlings were subjected to drought stress with the application of the NO donor sodium nitroprusside (SNP). A combination of confocal laser scanning microscopy, transcriptional expression analysis, biochemical assay of enzyme activity, virus-induced gene silencing (VIGS), and in vitro protein modification assays was applied to characterize the effects of NO on the drought stress response in cotton; (3) Results: Exogenous NO significantly reinforced drought resistance in cotton seedlings by improving leaf water retention capacity and photosynthetic efficiency, eliminating excessive drought-induced Reactive Oxygen Species (ROS), upregulating the transcription and enzymatic activity of antioxidant Enzymes, and promoting stomatal closure. Mechanistically, NO triggered S-nitrosylation of the plasma membrane H⁺-ATPase isoform GhHA2, thereby enhancing its protein stability; (4) Conclusions: These findings reveal that exogenous NO orchestrates cotton drought tolerance via multiple interconnected physiological and molecular pathways, in which the activation of the antioxidant defense system and the modulation of stomatal closure serve as central regulatory mechanisms.