Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Koiso, Narumi, Toda, Erika, Ichikawa, Masako, Kato, Norio, Okamoto, Takashi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2017
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
_version_ 1783417090374369280
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
work_keys_str_mv AT koisonarumi developmentofgeneexpressionsystemineggcellsandzygotesisolatedfromriceandmaize
AT todaerika developmentofgeneexpressionsystemineggcellsandzygotesisolatedfromriceandmaize
AT ichikawamasako developmentofgeneexpressionsystemineggcellsandzygotesisolatedfromriceandmaize
AT katonorio developmentofgeneexpressionsystemineggcellsandzygotesisolatedfromriceandmaize
AT okamototakashi developmentofgeneexpressionsystemineggcellsandzygotesisolatedfromriceandmaize