Single-cell transcriptomics and time-series metabolite profiling reveal the spatiotemporal regulation of flavonoid biosynthesis genes and phytohormone homeostasis by PAP1 in Arabidopsis

Abstract Background Understanding the spatiotemporal regulation of specialized metabolism in plants is critical for advancing both basic plant biology and biotechnological applications. PRODUCTION OF ANTHOCYANIN PIGMENT 1 (PAP1) is a well-known transcription factor that plays a key regulatory role i...

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Main Authors: Bingxu Zhang, Thomas Ka Yam Lam, Linheng Chen, Chen Zhang, Liping Zhu, Hailei Zhang, Pengxi Wang, Jianing Wang, Zongwei Cai, Yiji Xia
Format: Article
Language:English
Published: BMC 2025-07-01
Series:BMC Biology
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Online Access:https://doi.org/10.1186/s12915-025-02297-6
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author Bingxu Zhang
Thomas Ka Yam Lam
Linheng Chen
Chen Zhang
Liping Zhu
Hailei Zhang
Pengxi Wang
Jianing Wang
Zongwei Cai
Yiji Xia
author_facet Bingxu Zhang
Thomas Ka Yam Lam
Linheng Chen
Chen Zhang
Liping Zhu
Hailei Zhang
Pengxi Wang
Jianing Wang
Zongwei Cai
Yiji Xia
author_sort Bingxu Zhang
collection DOAJ
description Abstract Background Understanding the spatiotemporal regulation of specialized metabolism in plants is critical for advancing both basic plant biology and biotechnological applications. PRODUCTION OF ANTHOCYANIN PIGMENT 1 (PAP1) is a well-known transcription factor that plays a key regulatory role in the biosynthesis pathway of plant flavonoids. Similar to other secondary metabolites, flavonoid biosynthesis displays cell heterogeneity. However, the cell-specific regulation network of the flavonoid biosynthetic pathway remains unclear. Results In this study, we utilized single-cell RNA sequencing (scRNA-seq) and time-series metabolite profiling to investigate the regulation of flavonoid biosynthesis and phytohormone homeostasis in Arabidopsis thaliana by PAP1. By comparing single-cell transcriptomes of the pap1-D mutant and wild-type plant leaves, we constructed a cell-type-specific atlas of gene expression and high-resolution dynamics of metabolites across developmental stages. Our findings reveal that PAP1 overexpression induces distinct spatiotemporal regulation of phenylpropanoid pathway genes in different cell types and widespread upregulation of glycosylation processes. Metabolomic profiling validated these transcriptional patterns and showed significant changes of metabolites in phenylalanine metabolic processes as pap1-D leaf matures. Additionally, PAP1 overexpression leads to significant changes in phytohormone levels, particularly jasmonate and salicylate, indicating complex crosstalk between flavonoid biosynthesis and hormone homeostasis. Conclusions This integrated multi-omics approach provides unprecedented insights into the cell-specific regulatory networks controlling specialized metabolism and establishes a valuable framework for optimizing metabolic engineering strategies to enhance the production of bioactive plant compounds.
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spelling doaj-art-cf0b7f509aba49dea4965eec8dee8d8a2025-08-20T03:04:15ZengBMCBMC Biology1741-70072025-07-0123111910.1186/s12915-025-02297-6Single-cell transcriptomics and time-series metabolite profiling reveal the spatiotemporal regulation of flavonoid biosynthesis genes and phytohormone homeostasis by PAP1 in ArabidopsisBingxu Zhang0Thomas Ka Yam Lam1Linheng Chen2Chen Zhang3Liping Zhu4Hailei Zhang5Pengxi Wang6Jianing Wang7Zongwei Cai8Yiji Xia9Department of Biology, Hong Kong Baptist UniversityDepartment of Chemistry, Hong Kong Baptist UniversityDepartment of Biology, Hong Kong Baptist UniversityDepartment of Biology, Hong Kong Baptist UniversityDepartment of Biology, Hong Kong Baptist UniversityDepartment of Biology, Hong Kong Baptist UniversityDepartment of Biology, Hong Kong Baptist UniversityState Key Laboratory of Environmental and Biological Analysis, Hong Kong Baptist UniversityDepartment of Chemistry, Hong Kong Baptist UniversityDepartment of Biology, Hong Kong Baptist UniversityAbstract Background Understanding the spatiotemporal regulation of specialized metabolism in plants is critical for advancing both basic plant biology and biotechnological applications. PRODUCTION OF ANTHOCYANIN PIGMENT 1 (PAP1) is a well-known transcription factor that plays a key regulatory role in the biosynthesis pathway of plant flavonoids. Similar to other secondary metabolites, flavonoid biosynthesis displays cell heterogeneity. However, the cell-specific regulation network of the flavonoid biosynthetic pathway remains unclear. Results In this study, we utilized single-cell RNA sequencing (scRNA-seq) and time-series metabolite profiling to investigate the regulation of flavonoid biosynthesis and phytohormone homeostasis in Arabidopsis thaliana by PAP1. By comparing single-cell transcriptomes of the pap1-D mutant and wild-type plant leaves, we constructed a cell-type-specific atlas of gene expression and high-resolution dynamics of metabolites across developmental stages. Our findings reveal that PAP1 overexpression induces distinct spatiotemporal regulation of phenylpropanoid pathway genes in different cell types and widespread upregulation of glycosylation processes. Metabolomic profiling validated these transcriptional patterns and showed significant changes of metabolites in phenylalanine metabolic processes as pap1-D leaf matures. Additionally, PAP1 overexpression leads to significant changes in phytohormone levels, particularly jasmonate and salicylate, indicating complex crosstalk between flavonoid biosynthesis and hormone homeostasis. Conclusions This integrated multi-omics approach provides unprecedented insights into the cell-specific regulatory networks controlling specialized metabolism and establishes a valuable framework for optimizing metabolic engineering strategies to enhance the production of bioactive plant compounds.https://doi.org/10.1186/s12915-025-02297-6Single-cell transcriptomicsMetabolomicsPhytohormone homeostasisFlavonoidSpatiotemporal atlasArabidopsis
spellingShingle Bingxu Zhang
Thomas Ka Yam Lam
Linheng Chen
Chen Zhang
Liping Zhu
Hailei Zhang
Pengxi Wang
Jianing Wang
Zongwei Cai
Yiji Xia
Single-cell transcriptomics and time-series metabolite profiling reveal the spatiotemporal regulation of flavonoid biosynthesis genes and phytohormone homeostasis by PAP1 in Arabidopsis
BMC Biology
Single-cell transcriptomics
Metabolomics
Phytohormone homeostasis
Flavonoid
Spatiotemporal atlas
Arabidopsis
title Single-cell transcriptomics and time-series metabolite profiling reveal the spatiotemporal regulation of flavonoid biosynthesis genes and phytohormone homeostasis by PAP1 in Arabidopsis
title_full Single-cell transcriptomics and time-series metabolite profiling reveal the spatiotemporal regulation of flavonoid biosynthesis genes and phytohormone homeostasis by PAP1 in Arabidopsis
title_fullStr Single-cell transcriptomics and time-series metabolite profiling reveal the spatiotemporal regulation of flavonoid biosynthesis genes and phytohormone homeostasis by PAP1 in Arabidopsis
title_full_unstemmed Single-cell transcriptomics and time-series metabolite profiling reveal the spatiotemporal regulation of flavonoid biosynthesis genes and phytohormone homeostasis by PAP1 in Arabidopsis
title_short Single-cell transcriptomics and time-series metabolite profiling reveal the spatiotemporal regulation of flavonoid biosynthesis genes and phytohormone homeostasis by PAP1 in Arabidopsis
title_sort single cell transcriptomics and time series metabolite profiling reveal the spatiotemporal regulation of flavonoid biosynthesis genes and phytohormone homeostasis by pap1 in arabidopsis
topic Single-cell transcriptomics
Metabolomics
Phytohormone homeostasis
Flavonoid
Spatiotemporal atlas
Arabidopsis
url https://doi.org/10.1186/s12915-025-02297-6
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