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The phytohormone abscisic acid (ABA) induces stomatal closure in response to drought stress, leading to reduction of transpirational water loss. A thiol tripeptide glutathione (GSH) is an important regulator of cellular redox homeostasis in plants. Although it has been shown that cellular redox state of guard cells controls ABA-mediated stomatal closure, roles of GSH in guard cell ABA signaling were largely unknown. Recently we demonstrated that GSH functions as a negative regulator of ABA signaling in guard cells. In this study we performed more detailed analyses to reveal how GSH regulates guard cell ABA signaling using the GSH-deficient Arabidopsis mutant  cad2-1 . The  cad2-1  mutant exhibited reduced water loss from rosette leaves. Whole-cell current recording using patch clamp technique revealed that the  cad2-1  mutation did not affect ABA regulation of S-type anion channels. We found enhanced activation of Ca 2+  permeable channels by hydrogen peroxide (H 2 O 2 ) in  cad2-1  guard cells. The  cad2-1  mutant showed enhanced H 2 O 2 -induced stomatal closure and significant increase of ROS accumulation in whole leaves in response to ABA. Our findings provide a new understanding of guard cell ABA signaling and a new strategy to improve plant drought tolerance.

Regulation of reactive oxygen species-mediated abscisic acid signaling in guard cells and drought tolerance by glutathione