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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...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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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 |
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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 |
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