A persulfidation-based reversible post-translational modification of Cys desulfhydrase and NADPH oxidase RBOHD fine-tunes guard cell ABA signaling.

Hydrogen sulfide (H₂S) is an important signaling molecule that regulates diverse cellular signaling pathways through persulfidation. Our previous study revealed that H₂S is involved in the improvement of rice drought tolerance. However, the corresponding enzymatic sources of H₂S and its regulatory mechanism in response to drought stress are not clear. Here, we cloned and characterized a putative l-cysteine desulfhydrase (LCD) gene in rice, which encodes a protein possessing H₂S-producing activity and was named OsLCD1. Overexpression of OsLCD1 results in enhanced H₂S production, persulfidation of total soluble protein, and confers rice drought tolerance. Further, we found that nitrate reductase (NR) activity was decreased under drought stress, and the inhibition of NR activity was controlled by endogenous H₂S production. Persulfidation of NIA2, an NR isoform responsible for the main NR activity, led to a decrease in total NR activity in rice. Furthermore, drought stress-triggered inhibition of NR activity and persulfidation of NIA2 was intensified in the OsLCD1 overexpression line. Phenotypical and molecular analysis revealed that mutation of NIA2 enhanced rice drought tolerance by activating the expression of genes encoding antioxidant enzymes and ABA-responsive genes. Taken together, our results showed the role of OsLCD1 in modulating H₂S production and provided insight into H₂S-regulated persulfidation of NIA2 in the control of rice drought stress.

Hydrogen sulfide (H₂S) is an endogenous gaseous molecule that plays an important role in the plant life cycle. The multiple transcription factor ABSCISIC ACID INSENSITIVE 4 (ABI4) was precisely regulated to participate in the abscisic acid (ABA) mediated signaling cascade. However, the molecular mechanisms of how H₂S regulates ABI4 protein level to control seed germination and seedling growth have remained elusive. In this study, we demonstrated that ABI4 controls the expression of L-CYSTEINE DESULFHYDRASE1 (DES1), a critical endogenous H₂S-producing enzyme, and both ABI4 and DES1-produced H₂S have inhibitory effects on seed germination. Furthermore, the ABI4 level decreased during seed germination while H₂S triggered the enhancement of the persulfidation level of ABI4 and alleviated its degradation rate, which in turn inhibited seed germination and seedling establishment. Conversely, the mutation of ABI4 at Cys250 decreased ABI4 protein stability and facilitated seed germination. Moreover, ABI4 degradation is also regulated via the 26S proteasome pathway. Taken together, these findings suggest a molecular link between DES1 and ABI4 through the post-translational modifications of persulfidation during early seedling development.

Hydrogen sulfide (H₂S) is a versatile signaling molecule that regulates multiple physiological processes in plants, including growth and development, immunity, and stress response as well. Signaling triggered by H₂S is proposed to occur via persulfidation, an oxidative post-translational modification (PTM) of cysteine residues (–SH) to persulfides (–SSH). Notwithstanding the growing body of information for the plant persulfidation proteome, the gap between the molecular mechanism of H₂S and physiological functions of protein persulfidation remains large. In this mini-review, we discussed the specific regulatory mechanism of persulfidation on guard cell abscisic acid (ABA) signaling and the possible link of persulfidation, sulfenylation, and S-nitrosylation within the framework of redox-based regulation.