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Highly efficient Agrobacterium rhizogenes-mediated transformation for functional analysis in woodland strawberry
BACKGROUND: The diploid woodland strawberry (Fragaria vesca) is an excellent model plant for investigating economically significant traits and several genetic resources within the Rosaceae family. Agrobacterium rhizogenes-mediated hairy root transformation is an alternative for exploring gene functi...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10517450/ https://www.ncbi.nlm.nih.gov/pubmed/37742022 http://dx.doi.org/10.1186/s13007-023-01078-y |
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author | Yan, Huiqing Ma, Dandan Yi, Peipei Sun, Guilian Chen, Xingyan Yi, Yin Huang, Xiaolong |
author_facet | Yan, Huiqing Ma, Dandan Yi, Peipei Sun, Guilian Chen, Xingyan Yi, Yin Huang, Xiaolong |
author_sort | Yan, Huiqing |
collection | PubMed |
description | BACKGROUND: The diploid woodland strawberry (Fragaria vesca) is an excellent model plant for investigating economically significant traits and several genetic resources within the Rosaceae family. Agrobacterium rhizogenes-mediated hairy root transformation is an alternative for exploring gene functions, especially the genes specifically expressed in roots. However, the hairy root transformation has not been established in strawberry. RESULTS: Here, we described an efficient and rapid hairy root transgenic system for strawberry using A. rhizogenes. Strain of A. rhizogenes MSU440 or C58C1 was the most suitable for hairy root transformation. The transformation efficiency was highest when tissues contained hypocotyls as explants. The optimal procedure involves A. rhizogenes at an optical density (OD(600)) of 0.7 for 10 min and co-cultivation duration for four days, achieving a transgenic efficiency of up to 71.43%. An auxin responsive promoter DR5(ver2) carrying an enhanced green fluorescent protein (eGFP) marker was transformed by A. rhizogenes MSU440, thereby generating transgenic hairy roots capable of high eGFP expression in root tip and meristem of strawberry where auxin accumulated. Finally, this system was applied for functional analysis using jGCaMP7c, which could sense calcium signals. A significant upsurge in eGFP expression in the transgenic hairy roots was displayed after adding calcium chloride. The results suggested that this approach was feasible for studying specific promoters and could be a tool to analyze gene functions in the roots of strawberries. CONCLUSION: We established a rapid and efficient hairy root transformation in strawberry by optimizing parameters, which was adequate for promoter analysis and functional characterization of candidate genes in strawberry and other rosaceous plants. |
format | Online Article Text |
id | pubmed-10517450 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-105174502023-09-24 Highly efficient Agrobacterium rhizogenes-mediated transformation for functional analysis in woodland strawberry Yan, Huiqing Ma, Dandan Yi, Peipei Sun, Guilian Chen, Xingyan Yi, Yin Huang, Xiaolong Plant Methods Methodology BACKGROUND: The diploid woodland strawberry (Fragaria vesca) is an excellent model plant for investigating economically significant traits and several genetic resources within the Rosaceae family. Agrobacterium rhizogenes-mediated hairy root transformation is an alternative for exploring gene functions, especially the genes specifically expressed in roots. However, the hairy root transformation has not been established in strawberry. RESULTS: Here, we described an efficient and rapid hairy root transgenic system for strawberry using A. rhizogenes. Strain of A. rhizogenes MSU440 or C58C1 was the most suitable for hairy root transformation. The transformation efficiency was highest when tissues contained hypocotyls as explants. The optimal procedure involves A. rhizogenes at an optical density (OD(600)) of 0.7 for 10 min and co-cultivation duration for four days, achieving a transgenic efficiency of up to 71.43%. An auxin responsive promoter DR5(ver2) carrying an enhanced green fluorescent protein (eGFP) marker was transformed by A. rhizogenes MSU440, thereby generating transgenic hairy roots capable of high eGFP expression in root tip and meristem of strawberry where auxin accumulated. Finally, this system was applied for functional analysis using jGCaMP7c, which could sense calcium signals. A significant upsurge in eGFP expression in the transgenic hairy roots was displayed after adding calcium chloride. The results suggested that this approach was feasible for studying specific promoters and could be a tool to analyze gene functions in the roots of strawberries. CONCLUSION: We established a rapid and efficient hairy root transformation in strawberry by optimizing parameters, which was adequate for promoter analysis and functional characterization of candidate genes in strawberry and other rosaceous plants. BioMed Central 2023-09-23 /pmc/articles/PMC10517450/ /pubmed/37742022 http://dx.doi.org/10.1186/s13007-023-01078-y Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Methodology Yan, Huiqing Ma, Dandan Yi, Peipei Sun, Guilian Chen, Xingyan Yi, Yin Huang, Xiaolong Highly efficient Agrobacterium rhizogenes-mediated transformation for functional analysis in woodland strawberry |
title | Highly efficient Agrobacterium rhizogenes-mediated transformation for functional analysis in woodland strawberry |
title_full | Highly efficient Agrobacterium rhizogenes-mediated transformation for functional analysis in woodland strawberry |
title_fullStr | Highly efficient Agrobacterium rhizogenes-mediated transformation for functional analysis in woodland strawberry |
title_full_unstemmed | Highly efficient Agrobacterium rhizogenes-mediated transformation for functional analysis in woodland strawberry |
title_short | Highly efficient Agrobacterium rhizogenes-mediated transformation for functional analysis in woodland strawberry |
title_sort | highly efficient agrobacterium rhizogenes-mediated transformation for functional analysis in woodland strawberry |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10517450/ https://www.ncbi.nlm.nih.gov/pubmed/37742022 http://dx.doi.org/10.1186/s13007-023-01078-y |
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