ABA signalling and metabolism are not essential for dark-induced stomatal closure but affect response speed

Stomata are microscopic pores that open and close, acting to balance CO₂ uptake with water loss. Stomata close in response to various signals including the drought hormone abscisic acid (ABA), microbe-associated-molecular-patterns, high CO₂ levels, and darkness. The signalling pathways underlying ABA-induced stomatal closure are well known, however, the mechanism for dark-induced stomatal closure is less clear. ABA signalling has been suggested to play a role in dark-induced stomatal closure, but it is unclear how this occurs. Here we investigate the role of ABA in promoting dark-induced stomatal closure. Tracking stomatal movements on the surface of leaf discs we find, although steady state stomatal apertures are affected by mutations in ABA signalling and metabolism genes, all mutants investigated close in response to darkness. However, we observed a delayed response to darkness for certain ABA signalling and metabolism mutants. Investigating this further in the quadruple ABA receptor mutant (pyr1pyl1pyl2pyl4), compared with wild-type, we found reduced stomatal conductance kinetics. Although our results suggest a non-essential role for ABA in dark-induced stomatal closure, we show that ABA modulates the speed of the dark-induced closure response. These results highlight the role of ABA signalling and metabolic pathways as potential targets for enhancing stomatal movement kinetics.