A transcription factor ensemble orchestrates bundle sheath expression in rice

Abstract C4 photosynthesis has evolved in over sixty plant lineages and improves photosynthetic efficiency by ~50%. One unifying character of C4 plants is photosynthetic activation of a compartment such as the bundle sheath, but gene regulatory networks controlling this cell type are poorly understo...

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Main Authors: Lei Hua, Na Wang, Susan Stanley, Ruth M. Donald, Satish Kumar Eeda, Kumari Billakurthi, Ana Rita Borba, Julian M. Hibberd
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-62087-0
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author Lei Hua
Na Wang
Susan Stanley
Ruth M. Donald
Satish Kumar Eeda
Kumari Billakurthi
Ana Rita Borba
Julian M. Hibberd
author_facet Lei Hua
Na Wang
Susan Stanley
Ruth M. Donald
Satish Kumar Eeda
Kumari Billakurthi
Ana Rita Borba
Julian M. Hibberd
author_sort Lei Hua
collection DOAJ
description Abstract C4 photosynthesis has evolved in over sixty plant lineages and improves photosynthetic efficiency by ~50%. One unifying character of C4 plants is photosynthetic activation of a compartment such as the bundle sheath, but gene regulatory networks controlling this cell type are poorly understood. In Arabidopsis, a bipartite MYC-MYB transcription factor module restricts gene expression to these cells, but in grasses the regulatory logic allowing bundle sheath gene expression has not been defined. Using the global staple and C3 crop rice, we find that the SULFITE REDUCTASE promoter is sufficient for strong bundle sheath expression. This promoter encodes an intricate cis-regulatory logic with multiple activators and repressors acting combinatorially. Within this landscape we identify a distal cis-regulatory module (CRM) activated by an ensemble of transcription factors from the WRKY, G2-like, MYB-related, DOF, IDD and bZIP families. This module is necessary and sufficient to pattern gene expression to the rice bundle sheath. Oligomerisation of the CRM and fusion to core promoters containing Y-patches allow activity to be increased 220-fold. This CRM generates bundle sheath-specific expression in Arabidopsis indicating deep conservation in function between monocotyledons and dicotyledons. In summary, we identify an ancient, short, and tuneable CRM patterning expression to the bundle sheath that we anticipate will be useful for engineering this cell type in various crop species.
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spelling doaj-art-c5b65c643a35406aa2ed18e06e6bc81c2025-08-20T03:05:05ZengNature PortfolioNature Communications2041-17232025-07-0116111710.1038/s41467-025-62087-0A transcription factor ensemble orchestrates bundle sheath expression in riceLei Hua0Na Wang1Susan Stanley2Ruth M. Donald3Satish Kumar Eeda4Kumari Billakurthi5Ana Rita Borba6Julian M. Hibberd7Department of Plant Sciences, University of CambridgeDepartment of Plant Sciences, University of CambridgeDepartment of Plant Sciences, University of CambridgeDepartment of Plant Sciences, University of CambridgeDepartment of Plant Sciences, University of CambridgeDepartment of Plant Sciences, University of CambridgeDepartment of Plant Sciences, University of CambridgeDepartment of Plant Sciences, University of CambridgeAbstract C4 photosynthesis has evolved in over sixty plant lineages and improves photosynthetic efficiency by ~50%. One unifying character of C4 plants is photosynthetic activation of a compartment such as the bundle sheath, but gene regulatory networks controlling this cell type are poorly understood. In Arabidopsis, a bipartite MYC-MYB transcription factor module restricts gene expression to these cells, but in grasses the regulatory logic allowing bundle sheath gene expression has not been defined. Using the global staple and C3 crop rice, we find that the SULFITE REDUCTASE promoter is sufficient for strong bundle sheath expression. This promoter encodes an intricate cis-regulatory logic with multiple activators and repressors acting combinatorially. Within this landscape we identify a distal cis-regulatory module (CRM) activated by an ensemble of transcription factors from the WRKY, G2-like, MYB-related, DOF, IDD and bZIP families. This module is necessary and sufficient to pattern gene expression to the rice bundle sheath. Oligomerisation of the CRM and fusion to core promoters containing Y-patches allow activity to be increased 220-fold. This CRM generates bundle sheath-specific expression in Arabidopsis indicating deep conservation in function between monocotyledons and dicotyledons. In summary, we identify an ancient, short, and tuneable CRM patterning expression to the bundle sheath that we anticipate will be useful for engineering this cell type in various crop species.https://doi.org/10.1038/s41467-025-62087-0
spellingShingle Lei Hua
Na Wang
Susan Stanley
Ruth M. Donald
Satish Kumar Eeda
Kumari Billakurthi
Ana Rita Borba
Julian M. Hibberd
A transcription factor ensemble orchestrates bundle sheath expression in rice
Nature Communications
title A transcription factor ensemble orchestrates bundle sheath expression in rice
title_full A transcription factor ensemble orchestrates bundle sheath expression in rice
title_fullStr A transcription factor ensemble orchestrates bundle sheath expression in rice
title_full_unstemmed A transcription factor ensemble orchestrates bundle sheath expression in rice
title_short A transcription factor ensemble orchestrates bundle sheath expression in rice
title_sort transcription factor ensemble orchestrates bundle sheath expression in rice
url https://doi.org/10.1038/s41467-025-62087-0
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