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Highly efficient hairy root genetic transformation and applications in citrus

Highly efficient genetic transformation technology is greatly beneficial for crop gene function analysis and precision breeding. However, the most commonly used genetic transformation technology for woody plants, mediated by Agrobacterium tumefaciens, is time-consuming and inefficient, which limits...

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Autores principales: Ma, Haijie, Meng, Xinyue, Xu, Kai, Li, Min, Gmitter, Fred G., Liu, Ningge, Gai, Yunpeng, Huang, Suya, Wang, Min, Wang, Nian, Xu, Hairen, Liu, Jinhua, Sun, Xuepeng, Duan, Shuo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9647159/
https://www.ncbi.nlm.nih.gov/pubmed/36388468
http://dx.doi.org/10.3389/fpls.2022.1039094
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author Ma, Haijie
Meng, Xinyue
Xu, Kai
Li, Min
Gmitter, Fred G.
Liu, Ningge
Gai, Yunpeng
Huang, Suya
Wang, Min
Wang, Min
Wang, Nian
Xu, Hairen
Liu, Jinhua
Sun, Xuepeng
Duan, Shuo
author_facet Ma, Haijie
Meng, Xinyue
Xu, Kai
Li, Min
Gmitter, Fred G.
Liu, Ningge
Gai, Yunpeng
Huang, Suya
Wang, Min
Wang, Min
Wang, Nian
Xu, Hairen
Liu, Jinhua
Sun, Xuepeng
Duan, Shuo
author_sort Ma, Haijie
collection PubMed
description Highly efficient genetic transformation technology is greatly beneficial for crop gene function analysis and precision breeding. However, the most commonly used genetic transformation technology for woody plants, mediated by Agrobacterium tumefaciens, is time-consuming and inefficient, which limits its utility for gene function analysis. In this study, a simple, universal, and highly efficient genetic transformation technology mediated by A. rhizogenes K599 is described. This technology can be applied to multiple citrus genotypes, and only 2–8 weeks were required for the entire workflow. Genome-editing experiments were simultaneously conducted using 11 plasmids targeting different genomic positions and all corresponding transformants with the target knocked out were obtained, indicating that A. rhizogenes-mediated genome editing was highly efficient. In addition, the technology is advantageous for investigation of specific genes (such as ACD2) for obtaining “hard-to-get” transgenic root tissue. Furthermore, A. rhizogenes can be used for direct viral vector inoculation on citrus bypassing the requirement for virion enrichment in tobacco, which facilitates virus-induced gene silencing and virus-mediated gene expression. In summary, we established a highly efficient genetic transformation technology bypassing tissue culture in citrus that can be used for genome editing, gene overexpression, and virus-mediated gene function analysis. We anticipate that by reducing the cost, required workload, experimental period, and other technical obstacles, this genetic transformation technology will be a valuable tool for routine investigation of endogenous and exogenous genes in citrus.
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spelling pubmed-96471592022-11-15 Highly efficient hairy root genetic transformation and applications in citrus Ma, Haijie Meng, Xinyue Xu, Kai Li, Min Gmitter, Fred G. Liu, Ningge Gai, Yunpeng Huang, Suya Wang, Min Wang, Min Wang, Nian Xu, Hairen Liu, Jinhua Sun, Xuepeng Duan, Shuo Front Plant Sci Plant Science Highly efficient genetic transformation technology is greatly beneficial for crop gene function analysis and precision breeding. However, the most commonly used genetic transformation technology for woody plants, mediated by Agrobacterium tumefaciens, is time-consuming and inefficient, which limits its utility for gene function analysis. In this study, a simple, universal, and highly efficient genetic transformation technology mediated by A. rhizogenes K599 is described. This technology can be applied to multiple citrus genotypes, and only 2–8 weeks were required for the entire workflow. Genome-editing experiments were simultaneously conducted using 11 plasmids targeting different genomic positions and all corresponding transformants with the target knocked out were obtained, indicating that A. rhizogenes-mediated genome editing was highly efficient. In addition, the technology is advantageous for investigation of specific genes (such as ACD2) for obtaining “hard-to-get” transgenic root tissue. Furthermore, A. rhizogenes can be used for direct viral vector inoculation on citrus bypassing the requirement for virion enrichment in tobacco, which facilitates virus-induced gene silencing and virus-mediated gene expression. In summary, we established a highly efficient genetic transformation technology bypassing tissue culture in citrus that can be used for genome editing, gene overexpression, and virus-mediated gene function analysis. We anticipate that by reducing the cost, required workload, experimental period, and other technical obstacles, this genetic transformation technology will be a valuable tool for routine investigation of endogenous and exogenous genes in citrus. Frontiers Media S.A. 2022-10-27 /pmc/articles/PMC9647159/ /pubmed/36388468 http://dx.doi.org/10.3389/fpls.2022.1039094 Text en Copyright © 2022 Ma, Meng, Xu, Li, Gmitter, Liu, Gai, Huang, Wang, Wang, Wang, Xu, Liu, Sun and Duan 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
Ma, Haijie
Meng, Xinyue
Xu, Kai
Li, Min
Gmitter, Fred G.
Liu, Ningge
Gai, Yunpeng
Huang, Suya
Wang, Min
Wang, Min
Wang, Nian
Xu, Hairen
Liu, Jinhua
Sun, Xuepeng
Duan, Shuo
Highly efficient hairy root genetic transformation and applications in citrus
title Highly efficient hairy root genetic transformation and applications in citrus
title_full Highly efficient hairy root genetic transformation and applications in citrus
title_fullStr Highly efficient hairy root genetic transformation and applications in citrus
title_full_unstemmed Highly efficient hairy root genetic transformation and applications in citrus
title_short Highly efficient hairy root genetic transformation and applications in citrus
title_sort highly efficient hairy root genetic transformation and applications in citrus
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9647159/
https://www.ncbi.nlm.nih.gov/pubmed/36388468
http://dx.doi.org/10.3389/fpls.2022.1039094
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