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Development of gene expression system in egg cells and zygotes isolated from rice and maize
Polyethylene glycol calcium (PEG‐Ca(2+)) transfection‐mediated analysis allows rapid and efficient examination of gene function. To investigate the diverse cellular functions of genes of interest in plant cells, macromolecules, such as DNA, RNA, and proteins, are delivered into protoplasts prepared...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6508540/ https://www.ncbi.nlm.nih.gov/pubmed/31245659 http://dx.doi.org/10.1002/pld3.10 |
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author | Koiso, Narumi Toda, Erika Ichikawa, Masako Kato, Norio Okamoto, Takashi |
author_facet | Koiso, Narumi Toda, Erika Ichikawa, Masako Kato, Norio Okamoto, Takashi |
author_sort | Koiso, Narumi |
collection | PubMed |
description | Polyethylene glycol calcium (PEG‐Ca(2+)) transfection‐mediated analysis allows rapid and efficient examination of gene function. To investigate the diverse cellular functions of genes of interest in plant cells, macromolecules, such as DNA, RNA, and proteins, are delivered into protoplasts prepared from somatic tissues or calli using a PEG‐Ca(2+) transfection procedure. To take advantage of this macromolecule delivery system in the reproductive and developmental biology of angiosperms, this study established a PEG‐Ca(2+) transfection system with isolated egg cells and zygotes. The conditions for PEG and plasmid DNA concentrations for transfection of rice egg cells were first addressed, and ~30% of PEG‐Ca(2+)‐transfected egg cells showed exogenous and transient expressions of fluorescent proteins from plasmid DNA delivered into the cells. Interestingly, a dual expression of two different fluorescent proteins in the same egg cell using two kinds of plasmid DNAs was also observed. For PEG‐Ca(2+) transfection with maize zygotes, ~80% of zygotes showed expression of GFP proteins from plasmid DNA. Importantly, PEG‐transfected zygotes developed normally into cell masses and mature plants. These results suggest that the present PEG‐Ca(2+)‐mediated transient expression system provides a novel and effective platform for expressing and analyzing genes of interest in egg cells and zygotes. Moreover, combined with the CRISPR/Cas9 approach, the present transient expression system in zygotes will become a powerful and alternative tool for the preparation of gene‐edited plants. |
format | Online Article Text |
id | pubmed-6508540 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-65085402019-06-26 Development of gene expression system in egg cells and zygotes isolated from rice and maize Koiso, Narumi Toda, Erika Ichikawa, Masako Kato, Norio Okamoto, Takashi Plant Direct Original Articles Polyethylene glycol calcium (PEG‐Ca(2+)) transfection‐mediated analysis allows rapid and efficient examination of gene function. To investigate the diverse cellular functions of genes of interest in plant cells, macromolecules, such as DNA, RNA, and proteins, are delivered into protoplasts prepared from somatic tissues or calli using a PEG‐Ca(2+) transfection procedure. To take advantage of this macromolecule delivery system in the reproductive and developmental biology of angiosperms, this study established a PEG‐Ca(2+) transfection system with isolated egg cells and zygotes. The conditions for PEG and plasmid DNA concentrations for transfection of rice egg cells were first addressed, and ~30% of PEG‐Ca(2+)‐transfected egg cells showed exogenous and transient expressions of fluorescent proteins from plasmid DNA delivered into the cells. Interestingly, a dual expression of two different fluorescent proteins in the same egg cell using two kinds of plasmid DNAs was also observed. For PEG‐Ca(2+) transfection with maize zygotes, ~80% of zygotes showed expression of GFP proteins from plasmid DNA. Importantly, PEG‐transfected zygotes developed normally into cell masses and mature plants. These results suggest that the present PEG‐Ca(2+)‐mediated transient expression system provides a novel and effective platform for expressing and analyzing genes of interest in egg cells and zygotes. Moreover, combined with the CRISPR/Cas9 approach, the present transient expression system in zygotes will become a powerful and alternative tool for the preparation of gene‐edited plants. John Wiley and Sons Inc. 2017-09-06 /pmc/articles/PMC6508540/ /pubmed/31245659 http://dx.doi.org/10.1002/pld3.10 Text en © 2017 The Authors. Plant Direct published by American Society of Plant Biologists, Society for Experimental Biology and John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Articles Koiso, Narumi Toda, Erika Ichikawa, Masako Kato, Norio Okamoto, Takashi Development of gene expression system in egg cells and zygotes isolated from rice and maize |
title | Development of gene expression system in egg cells and zygotes isolated from rice and maize |
title_full | Development of gene expression system in egg cells and zygotes isolated from rice and maize |
title_fullStr | Development of gene expression system in egg cells and zygotes isolated from rice and maize |
title_full_unstemmed | Development of gene expression system in egg cells and zygotes isolated from rice and maize |
title_short | Development of gene expression system in egg cells and zygotes isolated from rice and maize |
title_sort | development of gene expression system in egg cells and zygotes isolated from rice and maize |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6508540/ https://www.ncbi.nlm.nih.gov/pubmed/31245659 http://dx.doi.org/10.1002/pld3.10 |
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