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An Easy and Rapid Transformation Protocol for Transient Expression in Cotton Fiber
Cotton fiber is the most important natural textile material in the world. Identification and functional characterization of genes regulating fiber development are fundamental for improving fiber quality and yield. However, stable cotton transformation is time-consuming, low in efficiency, and techni...
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/PMC8980934/ https://www.ncbi.nlm.nih.gov/pubmed/35392510 http://dx.doi.org/10.3389/fpls.2022.837994 |
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author | Shang, Xiaoguang Zhu, Lijie Duan, Yujia He, Qingfei Zhao, Meiyue Yu, Yujia Guo, Wangzhen |
author_facet | Shang, Xiaoguang Zhu, Lijie Duan, Yujia He, Qingfei Zhao, Meiyue Yu, Yujia Guo, Wangzhen |
author_sort | Shang, Xiaoguang |
collection | PubMed |
description | Cotton fiber is the most important natural textile material in the world. Identification and functional characterization of genes regulating fiber development are fundamental for improving fiber quality and yield. However, stable cotton transformation is time-consuming, low in efficiency, and technically complex. Moreover, heterologous systems, such as Arabidopsis and tobacco, did not always work to elucidate the function of cotton fiber specifically expressed genes or their promoters. For these reasons, constructing a rapid transformation system using cotton fibers is necessary to study fiber’s specifically expressed genes. In this study, we developed an easy and rapid Agrobacterium-mediated method for the transient transformation of genes and promoters in cotton fibers. First, we found that exogenous genes could be expressed in cotton fibers via using β-glucuronidase (GUS) and green fluorescence protein (GFP) as reporters. Second, parameters affecting transformation efficiency, including LBA4404 Agrobacterium strain, 3 h infection time, and 2-day incubation time, were determined. Third, four different cotton genes that are specifically expressed in fibers were transiently transformed in cotton fibers, and the transcripts of these genes were detected ten to thousand times increase over the control. Fourth, GUS staining and activity analysis demonstrated that the activity profiles of GhMYB212 and GhFSN1 promoters in transformed fibers are similar to their native activity in developmental fibers. Furthermore, the transient transformation method was confirmed to be suitable for subcellular localization studies. In summary, the presented Agrobacterium-mediated transient transformation method is a fast, simple, and effective system for promoter characterization and protein expression in cotton fibers. |
format | Online Article Text |
id | pubmed-8980934 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-89809342022-04-06 An Easy and Rapid Transformation Protocol for Transient Expression in Cotton Fiber Shang, Xiaoguang Zhu, Lijie Duan, Yujia He, Qingfei Zhao, Meiyue Yu, Yujia Guo, Wangzhen Front Plant Sci Plant Science Cotton fiber is the most important natural textile material in the world. Identification and functional characterization of genes regulating fiber development are fundamental for improving fiber quality and yield. However, stable cotton transformation is time-consuming, low in efficiency, and technically complex. Moreover, heterologous systems, such as Arabidopsis and tobacco, did not always work to elucidate the function of cotton fiber specifically expressed genes or their promoters. For these reasons, constructing a rapid transformation system using cotton fibers is necessary to study fiber’s specifically expressed genes. In this study, we developed an easy and rapid Agrobacterium-mediated method for the transient transformation of genes and promoters in cotton fibers. First, we found that exogenous genes could be expressed in cotton fibers via using β-glucuronidase (GUS) and green fluorescence protein (GFP) as reporters. Second, parameters affecting transformation efficiency, including LBA4404 Agrobacterium strain, 3 h infection time, and 2-day incubation time, were determined. Third, four different cotton genes that are specifically expressed in fibers were transiently transformed in cotton fibers, and the transcripts of these genes were detected ten to thousand times increase over the control. Fourth, GUS staining and activity analysis demonstrated that the activity profiles of GhMYB212 and GhFSN1 promoters in transformed fibers are similar to their native activity in developmental fibers. Furthermore, the transient transformation method was confirmed to be suitable for subcellular localization studies. In summary, the presented Agrobacterium-mediated transient transformation method is a fast, simple, and effective system for promoter characterization and protein expression in cotton fibers. Frontiers Media S.A. 2022-03-22 /pmc/articles/PMC8980934/ /pubmed/35392510 http://dx.doi.org/10.3389/fpls.2022.837994 Text en Copyright © 2022 Shang, Zhu, Duan, He, Zhao, Yu and Guo. 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 Shang, Xiaoguang Zhu, Lijie Duan, Yujia He, Qingfei Zhao, Meiyue Yu, Yujia Guo, Wangzhen An Easy and Rapid Transformation Protocol for Transient Expression in Cotton Fiber |
title | An Easy and Rapid Transformation Protocol for Transient Expression in Cotton Fiber |
title_full | An Easy and Rapid Transformation Protocol for Transient Expression in Cotton Fiber |
title_fullStr | An Easy and Rapid Transformation Protocol for Transient Expression in Cotton Fiber |
title_full_unstemmed | An Easy and Rapid Transformation Protocol for Transient Expression in Cotton Fiber |
title_short | An Easy and Rapid Transformation Protocol for Transient Expression in Cotton Fiber |
title_sort | easy and rapid transformation protocol for transient expression in cotton fiber |
topic | Plant Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8980934/ https://www.ncbi.nlm.nih.gov/pubmed/35392510 http://dx.doi.org/10.3389/fpls.2022.837994 |
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