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Establishment of a stable, effective and universal genetic transformation technique in the diverse species of Brassica oleracea

Brassica oleracea is an economically important species, including seven cultivated variants. Agrobacterium-mediated transformation of B. oleracea crops, mainly via hypocotyl and cotyledon, has been achieved in the past. However, previously established transformation methods showed low efficiency, se...

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Autores principales: Sheng, Xiaoguang, Yu, Huifang, Wang, Jiansheng, Shen, Yusen, Gu, Honghui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9597678/
https://www.ncbi.nlm.nih.gov/pubmed/36311069
http://dx.doi.org/10.3389/fpls.2022.1021669
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author Sheng, Xiaoguang
Yu, Huifang
Wang, Jiansheng
Shen, Yusen
Gu, Honghui
author_facet Sheng, Xiaoguang
Yu, Huifang
Wang, Jiansheng
Shen, Yusen
Gu, Honghui
author_sort Sheng, Xiaoguang
collection PubMed
description Brassica oleracea is an economically important species, including seven cultivated variants. Agrobacterium-mediated transformation of B. oleracea crops, mainly via hypocotyl and cotyledon, has been achieved in the past. However, previously established transformation methods showed low efficiency, severe genotype limitation and a prolonged period for transformants acquisition, greatly restricting its application in functional genomic studies and crop improvement. In this study, we have compared the shoot regeneration and genetic transformation efficiency of hypocotyl, cotyledon petiole and curd peduncle explants from twelve genotypes of cauliflower and broccoli. Finally, an Agrobacterium-mediated transformation method using curd peduncle as explant was established, which is rapid, efficient, and amenable to high-throughput transformation and genome editing. The average genetic transformation efficiency of this method is stable up to 11.87% and was successfully implemented in twelve different genotypes of cauliflower and broccoli and other B. oleracea crops with low genotype dependence. Peduncle explants were found to contain abundant cambial cells with a strong cell division and shoot regeneration ability, which might be why this method achieved stable and high genetic transformation efficiency with almost no genotype dependence.
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spelling pubmed-95976782022-10-27 Establishment of a stable, effective and universal genetic transformation technique in the diverse species of Brassica oleracea Sheng, Xiaoguang Yu, Huifang Wang, Jiansheng Shen, Yusen Gu, Honghui Front Plant Sci Plant Science Brassica oleracea is an economically important species, including seven cultivated variants. Agrobacterium-mediated transformation of B. oleracea crops, mainly via hypocotyl and cotyledon, has been achieved in the past. However, previously established transformation methods showed low efficiency, severe genotype limitation and a prolonged period for transformants acquisition, greatly restricting its application in functional genomic studies and crop improvement. In this study, we have compared the shoot regeneration and genetic transformation efficiency of hypocotyl, cotyledon petiole and curd peduncle explants from twelve genotypes of cauliflower and broccoli. Finally, an Agrobacterium-mediated transformation method using curd peduncle as explant was established, which is rapid, efficient, and amenable to high-throughput transformation and genome editing. The average genetic transformation efficiency of this method is stable up to 11.87% and was successfully implemented in twelve different genotypes of cauliflower and broccoli and other B. oleracea crops with low genotype dependence. Peduncle explants were found to contain abundant cambial cells with a strong cell division and shoot regeneration ability, which might be why this method achieved stable and high genetic transformation efficiency with almost no genotype dependence. Frontiers Media S.A. 2022-10-12 /pmc/articles/PMC9597678/ /pubmed/36311069 http://dx.doi.org/10.3389/fpls.2022.1021669 Text en Copyright © 2022 Sheng, Yu, Wang, Shen and Gu 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
Sheng, Xiaoguang
Yu, Huifang
Wang, Jiansheng
Shen, Yusen
Gu, Honghui
Establishment of a stable, effective and universal genetic transformation technique in the diverse species of Brassica oleracea
title Establishment of a stable, effective and universal genetic transformation technique in the diverse species of Brassica oleracea
title_full Establishment of a stable, effective and universal genetic transformation technique in the diverse species of Brassica oleracea
title_fullStr Establishment of a stable, effective and universal genetic transformation technique in the diverse species of Brassica oleracea
title_full_unstemmed Establishment of a stable, effective and universal genetic transformation technique in the diverse species of Brassica oleracea
title_short Establishment of a stable, effective and universal genetic transformation technique in the diverse species of Brassica oleracea
title_sort establishment of a stable, effective and universal genetic transformation technique in the diverse species of brassica oleracea
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9597678/
https://www.ncbi.nlm.nih.gov/pubmed/36311069
http://dx.doi.org/10.3389/fpls.2022.1021669
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