Water-saving GC-MC model captures temporally differential enzymatic and transporter activities during C3-CAM transition

CAM (Crassulacean Acid Metabolism) plants reduce the water loss through transpiration in arid environments (Wickell et al., 2021), using an alternative pathway of carbon assimilation. To ensure food security, engineering CAM into C 3 plants can be achieved by inverting the stomatal rhythm and the timing of major CO 2 uptake from day to night. Identification of the metabolic enzymes and intra-cellular transporters, present in both C 3 and CAM but having different differential temporal activities throughout the diel cycle (Yang et al., 2015; Heyduk et al., 2019) and quantitative estimations of the flux distributions along the biochemical trajectory of C 3 -to-CAM transition may help us to achieve the goal. Here, we simulate a constraint-based combined metabolic model of guard cell (GC) and mesophyll cell (MC), linking temporal fluctuations of temperature (T) and relative humidity (RH) throughout the diurnal cycle with osmolyte accumulation dependent stomatal opening, CO 2 uptake and transpirational water loss. Starting with C 3 metabolism, gradual increase in water-use efficiency (WUE) captures several known and new differential activities of metabolic enzymes, transporters, sugar-malate cycle etc. in GC and MC during C 3 -to-CAM transition.