Synthetic alleles to study MUTE-dependent molecular transitions in stomatal development

Stomatal abundance sets plants potential for gas exchange, impacting photosynthesis and transpiration and, thus, plant survival and growth. Stomata originate from cell lineages initiated by asymmetric divisions of protodermal cells, producing meristemoids that develop into guard cell pairs. The tran...

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Authors: Illescas Miranda, Jonatan, Saiz Pérez, Josué, de Marcos , Alberto, Fenoll Comes, María Carmen, Mena Marugan, María de la Montaña
Format: article
Publication Date:2025
Country:España
Institution:Universidad de Castilla-La Mancha
Repository:RUIdeRA. Repositorio Institucional de la UCLM
OAI Identifier:oai:ruidera.uclm.es:10578/46519
Online Access:https://doi.org/10.1111/ppl.70072
https://hdl.handle.net/10578/46519
Access Level:Open access
Keyword:MUTE
Stomatal development
Synthetic alleles
Transcription factors
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spelling Synthetic alleles to study MUTE-dependent molecular transitions in stomatal developmentIllescas Miranda, JonatanSaiz Pérez, Josuéde Marcos , AlbertoFenoll Comes, María CarmenMena Marugan, María de la MontañaMUTEStomatal developmentSynthetic allelesTranscription factorsStomatal abundance sets plants potential for gas exchange, impacting photosynthesis and transpiration and, thus, plant survival and growth. Stomata originate from cell lineages initiated by asymmetric divisions of protodermal cells, producing meristemoids that develop into guard cell pairs. The transcription factors SPEECHLESS, MUTE, and FAMA are essential for stomatal lineage development, sequentially driving cell division and differentiation events. Their absence produces stomataless epidermis, hindering analysis of their roles during lineage development. MUTE drives the transition from proliferating meristemoids to guard mother cells, committed to stomatal fate. We aim to explore the molecular mechanisms underlying MUTE activity, using partial loss-of-function alleles predicted to impair DNA-binding and to potentially alter MUTE transcriptional activity. We engineered mutant allele coding sequences, generated Arabidopsis lines carrying them and analyzed their epidermal and transcriptional phenotypes using microscopy and RNA-seq. Synthetic alleles driven by the MUTE promoter rescued the stomata less phenotype of the seedling-lethal mute-3 mutant, enabling stomata differentiation and resulting in viable, fertile plants. Further examination of the developmental consequences of MUTE partial loss-of-function revealed arrested lineages, reduced stomatal abundance and altered stomatal spacing. Transcriptomic analysis of very young cotyledons from complemented lines indicated that only some MUTE targets require an intact MUTE bHLH domain. Comparison with existing lineage cell-specific transcriptional profiles showed that lineage development in the mutant lines was delayed compared to the wild-type but followed similar gene networks. These synthetic alleles provide new insight into MUTE ability to accurately and timely specify stomata formation.Wiley202620262025info:eu-repo/semantics/articleapplication/pdfapplication/pdfhttps://doi.org/10.1111/ppl.70072https://hdl.handle.net/10578/46519reponame:RUIdeRA. Repositorio Institucional de la UCLMinstname:Universidad de Castilla-La ManchaInglésPDI2019-105362RB-100PID2022-137606NB-100SBPLY/180225/000082022-GRIN-34125info:eu-repo/semantics/openAccessoai:ruidera.uclm.es:10578/465192026-05-27T07:36:41Z
dc.title.none.fl_str_mv Synthetic alleles to study MUTE-dependent molecular transitions in stomatal development
title Synthetic alleles to study MUTE-dependent molecular transitions in stomatal development
spellingShingle Synthetic alleles to study MUTE-dependent molecular transitions in stomatal development
Illescas Miranda, Jonatan
MUTE
Stomatal development
Synthetic alleles
Transcription factors
title_short Synthetic alleles to study MUTE-dependent molecular transitions in stomatal development
title_full Synthetic alleles to study MUTE-dependent molecular transitions in stomatal development
title_fullStr Synthetic alleles to study MUTE-dependent molecular transitions in stomatal development
title_full_unstemmed Synthetic alleles to study MUTE-dependent molecular transitions in stomatal development
title_sort Synthetic alleles to study MUTE-dependent molecular transitions in stomatal development
dc.creator.none.fl_str_mv Illescas Miranda, Jonatan
Saiz Pérez, Josué
de Marcos , Alberto
Fenoll Comes, María Carmen
Mena Marugan, María de la Montaña
author Illescas Miranda, Jonatan
author_facet Illescas Miranda, Jonatan
Saiz Pérez, Josué
de Marcos , Alberto
Fenoll Comes, María Carmen
Mena Marugan, María de la Montaña
author_role author
author2 Saiz Pérez, Josué
de Marcos , Alberto
Fenoll Comes, María Carmen
Mena Marugan, María de la Montaña
author2_role author
author
author
author
dc.subject.none.fl_str_mv MUTE
Stomatal development
Synthetic alleles
Transcription factors
topic MUTE
Stomatal development
Synthetic alleles
Transcription factors
description Stomatal abundance sets plants potential for gas exchange, impacting photosynthesis and transpiration and, thus, plant survival and growth. Stomata originate from cell lineages initiated by asymmetric divisions of protodermal cells, producing meristemoids that develop into guard cell pairs. The transcription factors SPEECHLESS, MUTE, and FAMA are essential for stomatal lineage development, sequentially driving cell division and differentiation events. Their absence produces stomataless epidermis, hindering analysis of their roles during lineage development. MUTE drives the transition from proliferating meristemoids to guard mother cells, committed to stomatal fate. We aim to explore the molecular mechanisms underlying MUTE activity, using partial loss-of-function alleles predicted to impair DNA-binding and to potentially alter MUTE transcriptional activity. We engineered mutant allele coding sequences, generated Arabidopsis lines carrying them and analyzed their epidermal and transcriptional phenotypes using microscopy and RNA-seq. Synthetic alleles driven by the MUTE promoter rescued the stomata less phenotype of the seedling-lethal mute-3 mutant, enabling stomata differentiation and resulting in viable, fertile plants. Further examination of the developmental consequences of MUTE partial loss-of-function revealed arrested lineages, reduced stomatal abundance and altered stomatal spacing. Transcriptomic analysis of very young cotyledons from complemented lines indicated that only some MUTE targets require an intact MUTE bHLH domain. Comparison with existing lineage cell-specific transcriptional profiles showed that lineage development in the mutant lines was delayed compared to the wild-type but followed similar gene networks. These synthetic alleles provide new insight into MUTE ability to accurately and timely specify stomata formation.
publishDate 2025
dc.date.none.fl_str_mv 2025
2026
2026
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://doi.org/10.1111/ppl.70072
https://hdl.handle.net/10578/46519
url https://doi.org/10.1111/ppl.70072
https://hdl.handle.net/10578/46519
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv PDI2019-105362RB-100
PID2022-137606NB-100
SBPLY/180225/00008
2022-GRIN-34125
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv reponame:RUIdeRA. Repositorio Institucional de la UCLM
instname:Universidad de Castilla-La Mancha
instname_str Universidad de Castilla-La Mancha
reponame_str RUIdeRA. Repositorio Institucional de la UCLM
collection RUIdeRA. Repositorio Institucional de la UCLM
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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