The authors used computational simulations of plant cellular metabolism under historical atmospheric conditions to demonstrate that reduced CO₂ and increased aridity can drive the evolutionary transition from C₃ to CAM photosynthesis. Their results suggest that while future elevated CO₂ may favor a reversion to C₃-like behavior, drought consistently promotes CAM regardless of CO₂ or temperature, and a minimum O₂ level is required for nocturnal respiration in CAM.

Using a microfluidic valve rootchip, the study simultaneously tracked ROS and calcium dynamics in compressed roots and found three kinetic phases linking mechanosensitive channel activity, NADPH oxidase‑dependent ROS accumulation, and secondary calcium influx. Pharmacological inhibition revealed that a fast calcium response is mediated by plasma‑membrane mechanosensitive channels, while a slower calcium increase is driven by ROS production.

The study generated deep proteome and phosphoproteome datasets from guard cell‑enriched tissue to examine how phosphorylation regulates stomatal movements. Comparative analysis revealed increased phosphorylation of endomembrane trafficking and vacuolar proteins in closed stomata, supporting a role for phospho‑regulated trafficking in stomatal dynamics.

The study employed immunofluorescence labeling and fluorescence intensity quantification to examine tissue-specific cellular modifications in plants under drought stress, revealing targeted alterations in proteoglycans, polysaccharides, and AGPs in leaves and roots. These findings highlight the importance of in planta analyses for accurately capturing stress-induced structural changes.

The study used high-resolution chemical imaging to map cell-type specific metabolic changes in plant roots inoculated with a nine-strain endophyte consortium under drought, revealing that endophytes differentially alter root metabolomes across spatial domains. Machine learning identified metabolites and exudates predictive of drought and endophyte treatment, and correlation analyses showed dynamic endophyte–metabolite relationships under stress.

The study engineered plastid-targeted genetically encoded biosensors to visualize several phosphoinositide species (PI3P, PI4P, PI5P, PI(4,5)P2, PI(3,5)P2) within chloroplasts and confirmed their specificity via immunological assays. Co‑expression with PPI‑modifying enzymes revealed altered biosensor distribution, an association between PI3P and the chloroplast protein VIPP1, and that elevated PPI levels under stress change localization and increase drought sensitivity, highlighting PPIs in plant stress responses.