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<p><i>Pogostemon cablin</i> Benth. (<i>P. cablin</i>) is an annual aromatic medicinal plant. In this study, anthers were cultured in vitro during the microspore development stage of <i>P. cablin</i>, 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 <i>PcNAC048</i> is expressed in different tissues of <i>P. cablin</i> and responds to abiotic stress. Transgenic studies in <i>Arabidopsis thaliana</i> showed that <i>PcNAC048</i> can promote lateral root development. Results from yeast one<i>-</i>hybrid and dual luciferase assays showed that PcNAC048 can interact with the promoter of the patchouli alcohol synthase gene (<i>PcPTS</i>) and inhibit its activity. Transient overexpression and virus-induced gene silencing (VIGS) analysis further confirmed that <i>PcNAC048</i> can negatively regulate the biosynthesis of patchouli alcohol. Overall, this study provides theoretical support for germplasm development and regulation of medicinal compounds in <i>P. cablin</i>.</p>

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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

  • 期刊论文

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  • 发表日期：

    2025年12月29日

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  • Xiaobing Wang,
  • Xiaojin Ge,
  • Wenxuan Zhong,
  • Daidi Wu,
  • Zhiye Yang,
  • Ruoting Zhan,
  • Likai Chen,
  • Hua Li

摘要

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.