Abstract
Pogostemon cablin Benth. (P. cablin) is an annual aromatic medicinal plant. In this study, anthers were cultured in vitro during the microspore development stage of P. cablin, inducing somatic embryogenesis. Globular somatic embryos (GSE), heart-shaped somatic embryos, torpedo-shaped somatic embryos, and cotyledonary somatic embryos (CSE) were observed and isolated. Single-cell RNA sequencing was then employed to generate single-cell maps for GSE and CSE. Using reported marker genes, a total of eight cell types were identified. Pseudo-temporal analysis reconstructed the continuous differentiation trajectory of apical meristem cells and epidermal cells. Further investigation identified PcNAC048 as a putative transcription factor that regulates embryonic cell differentiation. Gene expression analysis showed PcNAC048 is expressed in different tissues of P. cablin and responds to abiotic stress. Transgenic studies in Arabidopsis thaliana showed that PcNAC048 can promote lateral root development. Results from yeast one-hybrid and dual luciferase assays showed that PcNAC048 can interact with the promoter of the patchouli alcohol synthase gene (PcPTS) and inhibit its activity. Transient overexpression and virus-induced gene silencing (VIGS) analysis further confirmed that PcNAC048 can negatively regulate the biosynthesis of patchouli alcohol. Overall, this study provides theoretical support for germplasm development and regulation of medicinal compounds in P. cablin.
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Data availability
The original contributions presented in the study are included in the article/supplementary material. The scRNA-seq data (GSE304316) was deposited in the GEO repository. Uncropped and unedited gel image for Fig. S4 was available as Fig. S7 in the Supplementary Information file. The numerical source data for charts is shown in Supplementary Data 1. Further inquiries can be directed to the corresponding author.
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Acknowledgements
This research was supported by the National Natural Science Foundation of China (No. 82373976), the Natural Science Foundation of Guangdong Province (No. 2023A1515030194), the Special Project for Rural Revitalization Strategy in Guangdong Province (No. 2024-NPY-00-041 and 2023-NBH-00-022), and the Guangdong Province Seed Industry Revitalization Action Project (No. 2025-NQD-21-001). We thank Home for Researchers editorial team (www.home-for-researchers.com) for language editing service.
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L.C., X.W., H.L. and R.Z. designed the research protocols; X.W., X.G., W.Z., D.W. and Z.Y. conducted the experiments; X.W. and D.W. analyzed the raw data; X.W. and X.G. wrote the manuscript. All authors have read the manuscript and agree to submit the final version.
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Wang, X., Ge, X., Zhong, W. et al. Construction of a single-cell transcriptome atlas for Pogostemon cablin embryoids reveals PcNAC048 as a dual regulator coordinating lateral root morphogenesis and patchouli alcohol biosynthesis. Commun Biol (2025). https://doi.org/10.1038/s42003-025-09434-5
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DOI: https://doi.org/10.1038/s42003-025-09434-5
