Proteome-wide Identification of Cysteine S-sulfenylation in Response to Low Temperature and Low-humidity Stress in Dendrobium huoshanense

Objective Dendrobium huoshanense is a renowned geo-authentic herb, and its excellent quality is closely correlated with the specific combined stress of low temperature and low humidity found in its native habitat. Methods In this study, we profiled the protein S-sulfenylation landscape of D. huoshanense under simulated low temperature and low humidity stress using Cys-Tag labeling coupled with Data-Independent Acquisition (DIA) mass spectrometry. Results Physiological analysis revealed a moderate accumulation of H₂O₂ at 12 h of stress, representing an early signaling phase. In total, 9,924 sulfenylation sites mapped to 6,953 proteins were identified. Following stress treatment, 226 sites showed differential modification, 202 of which were upregulated. Motif analysis demonstrated a significant enrichment of lysine and arginine residues flanking the modification sites. Functional enrichment analysis indicated that S-sulfenylation is primarily associated with vesicle trafficking, carbon metabolism, and amino acid biosynthesis. Conclusion This study characterizes the response of sulfenylated proteins to low temperature and low humidity stress in D. huoshanense , providing theoretical insights into the molecular mechanisms of plant redox regulation under abiotic stress.