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...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-12-01
|
| Series: | Frontiers in Plant Science |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2022.1048695/full |
| _version_ | 1811314560304414720 |
|---|---|
| author | Dmitry Miroshnichenko Dmitry Miroshnichenko Dmitry Miroshnichenko Vadim Timerbaev Vadim Timerbaev Vadim Timerbaev Anna Klementyeva Alexander Pushin Alexander Pushin Tatiana Sidorova Dmitry Litvinov Dmitry Litvinov Lubov Nazarova Lubov Nazarova Olga Shulga Olga Shulga Mikhail Divashuk Mikhail Divashuk Gennady Karlov Elena Salina Sergey Dolgov Sergey Dolgov |
| author_facet | Dmitry Miroshnichenko Dmitry Miroshnichenko Dmitry Miroshnichenko Vadim Timerbaev Vadim Timerbaev Vadim Timerbaev Anna Klementyeva Alexander Pushin Alexander Pushin Tatiana Sidorova Dmitry Litvinov Dmitry Litvinov Lubov Nazarova Lubov Nazarova Olga Shulga Olga Shulga Mikhail Divashuk Mikhail Divashuk Gennady Karlov Elena Salina Sergey Dolgov Sergey Dolgov |
| author_sort | Dmitry Miroshnichenko |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-13T11:14:49Z |
| format | Article |
| id | doaj.art-db34f8f81d1a47888e9c789bb52aee52 |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-04-13T11:14:49Z |
| publishDate | 2022-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-db34f8f81d1a47888e9c789bb52aee522022-12-22T02:49:01ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2022-12-011310.3389/fpls.2022.10486951048695CRISPR/Cas9-induced modification of the conservative promoter region of VRN-A1 alters the heading time of hexaploid bread wheatDmitry Miroshnichenko0Dmitry Miroshnichenko1Dmitry Miroshnichenko2Vadim Timerbaev3Vadim Timerbaev4Vadim Timerbaev5Anna Klementyeva6Alexander Pushin7Alexander Pushin8Tatiana Sidorova9Dmitry Litvinov10Dmitry Litvinov11Lubov Nazarova12Lubov Nazarova13Olga Shulga14Olga Shulga15Mikhail Divashuk16Mikhail Divashuk17Gennady Karlov18Elena Salina19Sergey Dolgov20Sergey Dolgov21Kurchatov Genomic Center — All-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaBranch of Institute of Bioorganic Chemistry RAS, Pushchino, RussiaAll-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaKurchatov Genomic Center — All-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaBranch of Institute of Bioorganic Chemistry RAS, Pushchino, RussiaAll-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaBranch of Institute of Bioorganic Chemistry RAS, Pushchino, RussiaBranch of Institute of Bioorganic Chemistry RAS, Pushchino, RussiaAll-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaBranch of Institute of Bioorganic Chemistry RAS, Pushchino, RussiaKurchatov Genomic Center — All-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaAll-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaKurchatov Genomic Center — All-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaAll-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaKurchatov Genomic Center — All-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaAll-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaKurchatov Genomic Center — All-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaAll-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaAll-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaInstitute of Cytology and Genetics, SB RAS, Novosibirsk, RussiaBranch of Institute of Bioorganic Chemistry RAS, Pushchino, RussiaAll-Russia Research Institute of Agricultural Biotechnology, Moscow, RussiaIn 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.https://www.frontiersin.org/articles/10.3389/fpls.2022.1048695/fullgenome editingmutationvernalizationVRN-1 geneTriticum aestivum L. |
| spellingShingle | Dmitry Miroshnichenko Dmitry Miroshnichenko Dmitry Miroshnichenko Vadim Timerbaev Vadim Timerbaev Vadim Timerbaev Anna Klementyeva Alexander Pushin Alexander Pushin Tatiana Sidorova Dmitry Litvinov Dmitry Litvinov Lubov Nazarova Lubov Nazarova Olga Shulga Olga Shulga Mikhail Divashuk Mikhail Divashuk Gennady Karlov Elena Salina Sergey Dolgov Sergey Dolgov CRISPR/Cas9-induced modification of the conservative promoter region of VRN-A1 alters the heading time of hexaploid bread wheat Frontiers in Plant Science genome editing mutation vernalization VRN-1 gene Triticum aestivum L. |
| 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 | genome editing mutation vernalization VRN-1 gene Triticum aestivum L. |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2022.1048695/full |
| work_keys_str_mv | AT dmitrymiroshnichenko crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT dmitrymiroshnichenko crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT dmitrymiroshnichenko crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT vadimtimerbaev crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT vadimtimerbaev crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT vadimtimerbaev crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT annaklementyeva crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT alexanderpushin crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT alexanderpushin crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT tatianasidorova crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT dmitrylitvinov crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT dmitrylitvinov crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT lubovnazarova crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT lubovnazarova crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT olgashulga crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT olgashulga crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT mikhaildivashuk crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT mikhaildivashuk crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT gennadykarlov crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT elenasalina crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT sergeydolgov crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat AT sergeydolgov crisprcas9inducedmodificationoftheconservativepromoterregionofvrna1alterstheheadingtimeofhexaploidbreadwheat |