Transcriptomic analysis reveals the functions of H₂S as a gasotransmitter independently of Cys in Arabidopsis

Numerous studies have revealed the gasotransmitter functions of hydrogen sulfide (H₂S) in various biological processes. However, the involvement of H₂S in sulfur metabolism and/or Cys synthesis makes its role as a signaling molecule ambiguous. The generation of endogenous H₂S in plants is closely related to the metabolism of Cys, which play roles in a variety of signaling pathway occurring in various cellular processes. Here, we found that exogenous H₂S fumigation and Cys treatment modulated the production rate and content of endogenous H₂S and Cys to various degrees. Furthermore, we provided comprehensive transcriptomic analysis to support the gasotransmitter role of H₂S besides as a substrate for Cys synthesis. Comparison of the differentially expressed genes (DEGs) between H₂S and Cys treated seedlings indicated that H₂S fumigation and Cys treatment caused different influences on gene profiles during seedlings development. A total of 261 genes were identified to respond to H₂S fumigation, among which 72 genes were co-regulated by Cys treatment. GO and KEGG enrichment analysis of the 189 genes, H₂S but not Cys regulated DEGs, indicated that these genes mainly involved in plant hormone signal transduction, plant-pathogen interaction, phenylpropanoid biosynthesis, and MAPK signaling pathway. Most of these genes encoded proteins having DNA binding and transcription factor activities that play roles in a variety of plant developmental and environmental responses. Many stress-responsive genes and some Ca²⁺ signal associated genes were also included. Consequently, H₂S regulated gene expression through its role as a gasotransmitter, rather than just as a substrate for Cys biogenesis, and these 189 genes were far more likely to function in H₂S signal transduction independently of Cys. Our data will provide insights for revealing and enriching H₂S signaling networks.