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Increasing stomata movement is beneficial to improve plant water use efficiency and drought resilience. Contradictory results indicate that aquaporins might regulate stomatal movement. Here, we tested whether the maize plasma membrane PIP2;5 aquaporin affects stomatal closure under water deficit, abscisic acid (ABA) or vapor pressure deficit (VPD) treatment in intact plants, detached leaves, or peeled epidermis. Transpiration, stomatal conductance (gs) and aperture, and reactive oxygen species (ROS) in stomatal complexes were studied in maize lines with increased or knocked down (KD) PIP2;5 gene expression. In well-watered conditions, the PIP2;5 overexpressing (OE) plants transpired more than wild types (WT), while no significant difference in transpiration was observed between pip2;5 KD and WT. Upon mild-water deficit or low ABA concentration treatments, transpiration and gs decreased more in PIP2;5 OE, and less in pip2;5 KD lines, in comparison with WTs. In detached epidermis, ABA treatment induced faster stomatal closing in PIP2;5 OE lines compared to WT, while pip2;5 KD stomata were ABA insensitive. These phenotypes were associated with guard cell ROS accumulation. Additionally, PIP2;5 is involved in the transpiration decrease observed under high VPD. These data indicate that maize PIP2;5 is a key actor increasing the sensitivity of stomatal closure to water deficit

The plasma membrane intrinsic protein PIP2;5 is the most highly expressed aquaporin in maize (Zea mays) roots. Here, we investigated how deregulation of PIP2;5 expression affects water relations and growth using maize overexpression (OE; B104 inbred) or knockout (KO; W22 inbred) lines. The hydraulic conductivity of the cortex cells of roots grown hydroponically was higher in PIP2;5 OE and lower in pip2;5 KO lines compared with the corresponding wild-type plants. While whole-root conductivity decreased in the KO lines compared to the wild type, no difference was observed in OE plants. This paradox was interpreted using the MECHA hydraulic model, which computes the radial flow of water within root sections. The model hints that the plasma membrane permeability of the cells is not radially uniform but that PIP2;5 may be saturated in cell layers with apoplastic barriers, i.e. the endodermis and exodermis, suggesting the presence of posttranslational mechanisms controlling the abundance of PIP in the plasma membrane in these cells. At the leaf level, where the PIP2;5 gene is weakly expressed in wildtype plants, the hydraulic conductance was higher in the PIP2;5 OE lines compared with the wild-type plants, whereas no difference was observed in the pip2;5 KO lines. The temporal trend of leaf elongation rate, used as a proxy for that of xylem water potential, was faster in PIP2;5 OE plants upon mild stress, but not in well-watered conditions, demonstrating that PIP2;5 may play a beneficial role in plant growth under specific conditions.