Mesophyll-Specific Circadian Dynamics of CAM Induction in the Ice Plant Unveiled by Single-Cell Transcriptomics

Mesophyll-Specific Circadian Dynamics of CAM Induction in the Ice Plant Unveiled by Single-Cell Transcriptomics | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Mesophyll-Specific Circadian Dynamics of CAM Induction in the Ice Plant Unveiled by Single-Cell Transcriptomics Matias Kirst, Noé Perron, Christopher Dervinis, Wendell Pereira, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3783761/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Crassulacean acid metabolism (CAM) is an evolutionary modification of the C3 photosynthetic carbon dioxide fixation pathway used by approximately 7% of terrestrial plants to live in drought-prone environments. Facultative CAM species, such as Mesembryanthemum crystallinum (common ice plant), possess the unique ability to switch from C3 to CAM photosynthesis in response to high-salinity and water-deficit stress. Here we characterized the environmentally-triggered transition from C3 to CAM in the ice plant using single nucleus RNA sequencing (snRNA-seq) to identify its putative regulators, supported by a novel high-quality assembled and annotated genome. Analysis of snRNA-seq datasets from ice plant leaves transitioning between C3 and CAM collected at dawn and dusk revealed substantial transcriptional changes in mesophyll cells at the onset of CAM induction. Notably, our findings identify mesophyll sub-cell types engaged in either CAM or C3 photosynthesis at dusk. Cell trajectory inference analysis reconstructed both 24-hour CAM and C3 cycles, enabling a direct comparison of gene expression profiles in these pathways. This comparative study uncovered divergent expression patterns of key circadian clock genes in CAM and C3 cell trajectories, pointing to a connection between circadian regulation and CAM induction. Biological sciences/Plant sciences/Photosynthesis Biological sciences/Molecular biology/Transcriptomics Biological sciences/Genetics/Gene expression Biological sciences/Genetics/Genomics/Transcriptomics Full Text Additional Declarations There is NO Competing Interest. Supplementary Files Supplementarytable1GenomeStats.xlsx Dataset 1 Supplementarytable2Orthologs.xlsx Dataset 2 Supplementarytable3top50genespercluster.xlsx Dataset 3 Supplementarytable4DEGtreatments.xlsx Dataset 4 Supplementarytable5DEGcluster5vs6.xlsx Dataset 5 Supplementarytable6DEGtrajectories.xlsx Dataset 6 Supplementarytable7GeneDifferenceAnalyses.xlsx Dataset 7 Supplementarytable8IcePlantChloroplastGenes.xlsx Dataset 8 Supplementarytable9QCMetrics.xlsx Dataset 9 SupplementaryFigures.pdf Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3783761","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":262044235,"identity":"d1736400-2eab-443f-a6a1-2e418f9211bc","order_by":0,"name":"Matias 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