Hydrogen sulfide (H₂S), a gasotransmitter molecule, plays critical roles in stomatal closure and cellular bioenergetics. Alternative splicing (AS) is a key regulatory mechanism during plant development and stress responses; however, the interplay between H₂S signaling and AS in drought tolerance remains unexplored in Chinese cabbage. In this study, we found that the mitochondrial inner membrane enzyme succinate dehydrogenase (SDH) responds to H₂S signaling during stomatal closure. Silencing of BrSDH1-1 impaired the effects of H₂S on stomatal closure, SDH activity, and ATP production. RNA-Seq analysis revealed that H₂S modulates the AS of BrSDH1-1, resulting in transcript variants with differential expression. Overexpression of BrSDH1-1A and BrSDH1-1C in Arabidopsis enhanced drought resistance, whereas BrSDH1-1B had no significant effect. H₂S enhanced SDH activity and ATP production, promoted stomatal closure, and reduced excess reactive oxygen species (ROS) in OE-BrSDH1-1A and OE-BrSDH1-1C lines but not in OE-BrSDH1-1B. Furthermore, biotin-switch assays demonstrated that H₂S induced persulfidation of BrSDH1-1A and BrSDH1-1C, with no effect on variant BrSDH1-1B. These findings reveal a novel regulatory mechanism by which H₂S modulates BrSDH1-1 splicing to mediate stomatal closure and improve drought tolerance, offering valuable molecular insights for enhancing stress resilience in horticultural crops.

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