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Dual and spatially resolved drought responses in the Arabidopsis leaf mesophyll revealed by single-cell transcriptomics

ABSTRACT: Drought stress imposes severe challenges on agriculture by impacting crop performance. Understanding drought responses in plants at a cellular level is a crucial first step towards engineering improved drought resilience. The molecular responses to drought are however very complex as they depend on multiple factors including the severity of drought, the profiled organ, its developmental stage and even the cell types therein. As such, deciphering the transcriptional responses to drought is specifically challenging.In the study associated with this data submission, we investigated tissue-specific responses to mild drought in young Arabidopsis thaliana (Arabidopsis) leaves using single-cell RNA sequencing (scRNA-seq, deposited under another id, GSE273033). To preserve transcriptional integrity during cell isolation, we fixed the RNA prior to cell isolation using the transcription inhibitor actinomycin D, demonstrating the benefits of fixation for studying mild stress responses at single-cell level. The transcriptome data deposited here corresponds to the bulk RNA-sequencing experiments described in this study. The samples presented here are undigested leaves, leaves digested by cell wall degradation without fixation, and leaves digested by cell wall degradation with RNA fixation. All sample types were collected from plants growing under well-watered and mild drought conditions.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE273926 | GEO | 2025/01/16

REPOSITORIES: GEO

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Project description:Brachypodium distachyon as an annual species that colonises broken ground is a highly appropriate model to define drought tolerance mechanisms. We have previously identified accessions exhibiting drought tolerance, high susceptibility and intermediate tolerance to drought; respectively, ABR8, KOZ1 and ABR4, from a screen of 138 genotypes. To define the mechanisms of tolerance, the responses to drought were assessed using transcriptomic and metabolomic approaches. Analysis of RNA-seq before and following drought suggested relatively few differentially expressed genes in ABR8. Linking these to gene ontology (GO) terms revealed an enrichment for biological processes related to “regulated stress response”, “plant cell wall” and “oxidative stress”. Interestingly, drought tolerance also appeared to correlate with pre-existing differences in gene expression linked to GO terms associated with the cell wall. These included glycoside hydrolases involved in cell wall remodelling, pectin methylesterases, expansins and a pectin acetylesterase. Metabolomic assessments of the same samples also indicated few statistically significant changes in ABR8 with drought. Instead, pre-existing differences in the cell-wall associated metabolites appeared to correlate with drought tolerance. Our data suggests two different modes/levels at which cell wall characteristics can play a role in conferring drought tolerance: (i) an active response mode/level which involves stress induced changes in cell wall features to help the plant cope with drought and (ii) an intrinsic mode in which innate differences in cell wall composition and architecture between genotypes are important in determining tolerance to drought stress. Both modes seem to contribute to the drought tolerance of ABR8. Identification of the exact mechanisms through which the cell wall confers drought tolerance will be important to fully exploit the contribution of the cell wall in the development of drought tolerant cereals and other grasses.

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Dual and spatially resolved drought responses in the Arabidopsis leaf mesophyll revealed by single-cell transcriptomics

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