Light-induced stomatal opening in Arabidopsis is negatively regulated by chloroplast-originated OPDA signaling

Stomatal movement is orchestrated by diverse signaling cascades and metabolic activities in guard cells. Light triggers the opening of the pores through the phototropin-mediated pathway, which leads to the activation of plasma membrane H⁺-ATPase and thereby facilitates potassium accumulation through K_in⁺ channels. However, it remains poorly understood how phototropin signaling is fine-tuned to prevent excessive stomatal opening and consequent water loss. Here, we show that the stomatal response to light is negatively regulated by 12-oxo-phytodienoic acid (OPDA), an oxylipin metabolite produced through enzymatic oxygenation of polyunsaturated fatty acids (PUFAs). We identify a set of phospholipase-encoding genes, phospholipase (PLIP)1/2/3, which are transactivated rapidly in guard cells upon illumination in a phototropin-dependent manner. These phospholipases release PUFAs from the chloroplast membrane, which is oxidized by guard-cell lipoxygenases and further metabolized to OPDA. The OPDA-deficient mutants had wider stomatal pores, whereas mutants containing elevated levels of OPDA showed the opposite effect on stomatal aperture. Transmembrane solute fluxes that drive stomatal aperture were enhanced in lox6-1 guard cells, indicating that OPDA signaling ultimately impacts on activities of proton pumps and K_in⁺ channels. Interestingly, the accelerated stomatal kinetics in lox6-1 leads to increased plant growth without cost in water or macronutrient use. Together, our results reveal a new role for chloroplast membrane oxylipin metabolism in stomatal regulation. Moreover, the accelerated stomatal opening kinetics in OPDA-deficient mutants benefits plant growth and water use efficiency.