Single-nucleus RNA sequencing reveals a spatiotemporal pattern of H2S signaling in Chinese cabbage

This study is reported by Zhuping Jin's group from Shanxi University and Yanping Long's group from Southern University of Science and Technology. Plants rely on gasotransmitters like hydrogen sulfide (H₂S) to coordinate stress adaptation, yet how H₂S orchestrates spatiotemporal responses across cell types remains unknown. Traditional bulk RNA sequencing averages millions of cells, masking individual contributions. Single-cell transcriptomics offers a transformative solution. Jiao Zhang, Yang Chen, Bo Liu, and their colleagues determined to employ single-nucleus RNA sequencing (snRNA-seq) to decipher this puzzle in Chinese cabbage leaves.

The team generated a high-resolution snRNA-seq atlas under H₂S fumigation, capturing dynamic responses across 0, 0.5, 6, and 24 hours. They identified 18 distinct clusters corresponding to 9 major cell types. Notably, Cluster 4 was markedly enriched in H₂S-fumigated samples and exhibited high expression of plant immunity-related genes (e.g., WRKY33 and MKK4). RNA in situ hybridization confirmed this cluster as a stress-responsive mesophyll subpopulation.

H₂S induces globally coordinated transcriptional reprogramming, with over 40% of upregulated genes activated in at least two cell types. Notably, some genes transitioned from cell-type-restricted to broadly expressed patterns, exemplified by PYL4, a core ABA receptor, whose complex temporal dynamics reflected H₂S-mediated transcriptional reprogramming.

Two key genes enhancing drought tolerance were identified. BrHSP23.6, initially guard cell-specific, became globally expressed under H₂S and is required for photosynthetic enhancement and drought tolerance. Intriguingly, this protein lacks cysteine—the canonical persulfidation target—but contains six methionine residues. The researchers noted that confirmation of methionine persulfidation would expand the known scope of H₂S-mediated signaling.

BrNTF2, a de novo induced nuclear transport factor, is essential for H₂S-dependent stomatal regulation, functioning as a central hub in drought adaptation. Together, these genes operate through complementary non-stomatal and stomatal pathways to confer robust drought tolerance.

Further analyses revealed that H₂S spatiotemporally activates hormone and sulfur metabolic pathways in a layered manner, providing a cell-resolved framework for stress signaling. Given the conservation of core H₂S components, this spatiotemporal reprogramming pattern may extend across species and stresses, redefining H₂S as a spatiotemporal orchestrator of plant adaptation. The research team has deposited the data in the China National Center for Bioinformation (accession: PRJCA039764) and developed an online data browser (https://zhailab.bio.sustech.edu.cn/H2S_brassica), offering a valuable single-cell transcriptomic resource for the global research community.

 

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Single-nucleus RNA sequencing reveals a spatiotemporal pattern of H₂S signaling in Chinese cabbage

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