Cargando…

CRISPR/Cas9-induced modification of the conservative promoter region of VRN-A1 alters the heading time of hexaploid bread wheat

In cereals, the vernalization-related gene network plays an important role in regulating the transition from the vegetative to the reproductive phase to ensure optimal reproduction in a temperate climate. In hexaploid bread wheat (Triticum aestivum L.), the spring growth habit is associated with the...

Descripción completa

Detalles Bibliográficos
Autores principales: Miroshnichenko, Dmitry, Timerbaev, Vadim, Klementyeva, Anna, Pushin, Alexander, Sidorova, Tatiana, Litvinov, Dmitry, Nazarova, Lubov, Shulga, Olga, Divashuk, Mikhail, Karlov, Gennady, Salina, Elena, Dolgov, Sergey
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9760837/
https://www.ncbi.nlm.nih.gov/pubmed/36544871
http://dx.doi.org/10.3389/fpls.2022.1048695
_version_ 1784852570040172544
author Miroshnichenko, Dmitry
Timerbaev, Vadim
Klementyeva, Anna
Pushin, Alexander
Sidorova, Tatiana
Litvinov, Dmitry
Nazarova, Lubov
Shulga, Olga
Divashuk, Mikhail
Karlov, Gennady
Salina, Elena
Dolgov, Sergey
author_facet Miroshnichenko, Dmitry
Timerbaev, Vadim
Klementyeva, Anna
Pushin, Alexander
Sidorova, Tatiana
Litvinov, Dmitry
Nazarova, Lubov
Shulga, Olga
Divashuk, Mikhail
Karlov, Gennady
Salina, Elena
Dolgov, Sergey
author_sort Miroshnichenko, Dmitry
collection PubMed
description In cereals, the vernalization-related gene network plays an important role in regulating the transition from the vegetative to the reproductive phase to ensure optimal reproduction in a temperate climate. In hexaploid bread wheat (Triticum aestivum L.), the spring growth habit is associated with the presence of at least one dominant locus of VERNALIZATION 1 gene (VRN-1), which usually differs from recessive alleles due to mutations in the regulatory sequences of the promoter or/and the first intron. VRN-1 gene is a key regulator of floral initiation; various combinations of dominant and recessive alleles, especially VRN-A1 homeologs, determine the differences in the timing of wheat heading/flowering. In the present study, we attempt to expand the types of VRN-A1 alleles using CRISPR/Cas9 targeted modification of the promoter sequence. Several mono- and biallelic changes were achieved within the 125-117 bp upstream sequence of the start codon of the recessive vrn-A1 gene in plants of semi-winter cv. ‘Chinese Spring’. New mutations stably inherited in subsequent progenies and transgene-free homozygous plants carrying novel VRN-A1 variants were generated. Minor changes in the promoter sequence, such as 1–4 nucleotide insertions/deletions, had no effect on the heading time of plants, whereas the CRISPR/Cas9-mediated 8 bp deletion between −125 and −117 bp of the vrn-A1 promoter shortened the time of head emergence by up to 2-3 days. Such a growth habit was consistently observed in homozygous mutant plants under nonvernalized cultivation using different long day regimes (16, 18, or 22 h), whereas the cold treatment (from two weeks and more) completely leveled the effect of the 8 bp deletion. Importantly, comparison with wild-type plants showed that the implemented alteration has no negative effects on main yield characteristics. Our results demonstrate the potential to manipulate the heading time of wheat through targeted editing of the VRN-A1 gene promoter sequence on an otherwise unchanged genetic background.
format Online
Article
Text
id pubmed-9760837
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-97608372022-12-20 CRISPR/Cas9-induced modification of the conservative promoter region of VRN-A1 alters the heading time of hexaploid bread wheat Miroshnichenko, Dmitry Timerbaev, Vadim Klementyeva, Anna Pushin, Alexander Sidorova, Tatiana Litvinov, Dmitry Nazarova, Lubov Shulga, Olga Divashuk, Mikhail Karlov, Gennady Salina, Elena Dolgov, Sergey Front Plant Sci Plant Science In cereals, the vernalization-related gene network plays an important role in regulating the transition from the vegetative to the reproductive phase to ensure optimal reproduction in a temperate climate. In hexaploid bread wheat (Triticum aestivum L.), the spring growth habit is associated with the presence of at least one dominant locus of VERNALIZATION 1 gene (VRN-1), which usually differs from recessive alleles due to mutations in the regulatory sequences of the promoter or/and the first intron. VRN-1 gene is a key regulator of floral initiation; various combinations of dominant and recessive alleles, especially VRN-A1 homeologs, determine the differences in the timing of wheat heading/flowering. In the present study, we attempt to expand the types of VRN-A1 alleles using CRISPR/Cas9 targeted modification of the promoter sequence. Several mono- and biallelic changes were achieved within the 125-117 bp upstream sequence of the start codon of the recessive vrn-A1 gene in plants of semi-winter cv. ‘Chinese Spring’. New mutations stably inherited in subsequent progenies and transgene-free homozygous plants carrying novel VRN-A1 variants were generated. Minor changes in the promoter sequence, such as 1–4 nucleotide insertions/deletions, had no effect on the heading time of plants, whereas the CRISPR/Cas9-mediated 8 bp deletion between −125 and −117 bp of the vrn-A1 promoter shortened the time of head emergence by up to 2-3 days. Such a growth habit was consistently observed in homozygous mutant plants under nonvernalized cultivation using different long day regimes (16, 18, or 22 h), whereas the cold treatment (from two weeks and more) completely leveled the effect of the 8 bp deletion. Importantly, comparison with wild-type plants showed that the implemented alteration has no negative effects on main yield characteristics. Our results demonstrate the potential to manipulate the heading time of wheat through targeted editing of the VRN-A1 gene promoter sequence on an otherwise unchanged genetic background. Frontiers Media S.A. 2022-12-05 /pmc/articles/PMC9760837/ /pubmed/36544871 http://dx.doi.org/10.3389/fpls.2022.1048695 Text en Copyright © 2022 Miroshnichenko, Timerbaev, Klementyeva, Pushin, Sidorova, Litvinov, Nazarova, Shulga, Divashuk, Karlov, Salina and Dolgov https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Miroshnichenko, Dmitry
Timerbaev, Vadim
Klementyeva, Anna
Pushin, Alexander
Sidorova, Tatiana
Litvinov, Dmitry
Nazarova, Lubov
Shulga, Olga
Divashuk, Mikhail
Karlov, Gennady
Salina, Elena
Dolgov, Sergey
CRISPR/Cas9-induced modification of the conservative promoter region of VRN-A1 alters the heading time of hexaploid bread wheat
title CRISPR/Cas9-induced modification of the conservative promoter region of VRN-A1 alters the heading time of hexaploid bread wheat
title_full CRISPR/Cas9-induced modification of the conservative promoter region of VRN-A1 alters the heading time of hexaploid bread wheat
title_fullStr CRISPR/Cas9-induced modification of the conservative promoter region of VRN-A1 alters the heading time of hexaploid bread wheat
title_full_unstemmed CRISPR/Cas9-induced modification of the conservative promoter region of VRN-A1 alters the heading time of hexaploid bread wheat
title_short CRISPR/Cas9-induced modification of the conservative promoter region of VRN-A1 alters the heading time of hexaploid bread wheat
title_sort crispr/cas9-induced modification of the conservative promoter region of vrn-a1 alters the heading time of hexaploid bread wheat
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9760837/
https://www.ncbi.nlm.nih.gov/pubmed/36544871
http://dx.doi.org/10.3389/fpls.2022.1048695
work_keys_str_mv AT miroshnichenkodmitry crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT timerbaevvadim crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT klementyevaanna crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT pushinalexander crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT sidorovatatiana crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT litvinovdmitry crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT nazarovalubov crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT shulgaolga crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT divashukmikhail crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT karlovgennady crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT salinaelena crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat
AT dolgovsergey crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat