Cargando…
An asymmetric protoplast fusion and screening method for generating celeriac cybrids
Celeriac F(1) hybrid seed production is currently complicated due to the instability of cytoplasmic male sterile lines. To develop alternative alloplasmic CMS lines, an asymmetric protoplast fusion and hybrid screening methodology was established. Celeriac suspension cells protoplasts were used as t...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7907277/ https://www.ncbi.nlm.nih.gov/pubmed/33633203 http://dx.doi.org/10.1038/s41598-021-83970-y |
_version_ | 1783655465373138944 |
---|---|
author | Bruznican, Silvia Eeckhaut, Tom Van Huylenbroeck, Johan De Keyser, Ellen De Clercq, Hervé Geelen, Danny |
author_facet | Bruznican, Silvia Eeckhaut, Tom Van Huylenbroeck, Johan De Keyser, Ellen De Clercq, Hervé Geelen, Danny |
author_sort | Bruznican, Silvia |
collection | PubMed |
description | Celeriac F(1) hybrid seed production is currently complicated due to the instability of cytoplasmic male sterile lines. To develop alternative alloplasmic CMS lines, an asymmetric protoplast fusion and hybrid screening methodology was established. Celeriac suspension cells protoplasts were used as the acceptor and carrot, coriander and white celery mesophyll protoplasts as the donor for protoplast fusion experiments. Acceptor cytoplasmic inheritance was inhibited by iodoacetamide treatment and donor nuclear genome inheritance was prevented by UV exposure. Protoplasts were selectively stained and fused using electroporation and polyethylene glycol, and candidate hybrid shoots were obtained. One chloroplast and three mitochondrial markers that could distinguish acceptor and donors organelles were used to characterize over 600 plants obtained after fusion events, without identifying any cybrid. In order to increase the testing efficiency a high number of micro plantlets were pooled and hence the presence of the carrot specific Atp1 marker in one of the pooled samples was detected. We demonstrated that fusion took place between celeriac and a carrot indicating that the creation of viable hybrids is possible although at a very low frequency. These findings open the path for new cytoplasmic hybridization and the isolation of novel CMS lines of celeriac. |
format | Online Article Text |
id | pubmed-7907277 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-79072772021-03-02 An asymmetric protoplast fusion and screening method for generating celeriac cybrids Bruznican, Silvia Eeckhaut, Tom Van Huylenbroeck, Johan De Keyser, Ellen De Clercq, Hervé Geelen, Danny Sci Rep Article Celeriac F(1) hybrid seed production is currently complicated due to the instability of cytoplasmic male sterile lines. To develop alternative alloplasmic CMS lines, an asymmetric protoplast fusion and hybrid screening methodology was established. Celeriac suspension cells protoplasts were used as the acceptor and carrot, coriander and white celery mesophyll protoplasts as the donor for protoplast fusion experiments. Acceptor cytoplasmic inheritance was inhibited by iodoacetamide treatment and donor nuclear genome inheritance was prevented by UV exposure. Protoplasts were selectively stained and fused using electroporation and polyethylene glycol, and candidate hybrid shoots were obtained. One chloroplast and three mitochondrial markers that could distinguish acceptor and donors organelles were used to characterize over 600 plants obtained after fusion events, without identifying any cybrid. In order to increase the testing efficiency a high number of micro plantlets were pooled and hence the presence of the carrot specific Atp1 marker in one of the pooled samples was detected. We demonstrated that fusion took place between celeriac and a carrot indicating that the creation of viable hybrids is possible although at a very low frequency. These findings open the path for new cytoplasmic hybridization and the isolation of novel CMS lines of celeriac. Nature Publishing Group UK 2021-02-25 /pmc/articles/PMC7907277/ /pubmed/33633203 http://dx.doi.org/10.1038/s41598-021-83970-y Text en © The Author(s) 2021 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/. |
spellingShingle | Article Bruznican, Silvia Eeckhaut, Tom Van Huylenbroeck, Johan De Keyser, Ellen De Clercq, Hervé Geelen, Danny An asymmetric protoplast fusion and screening method for generating celeriac cybrids |
title | An asymmetric protoplast fusion and screening method for generating celeriac cybrids |
title_full | An asymmetric protoplast fusion and screening method for generating celeriac cybrids |
title_fullStr | An asymmetric protoplast fusion and screening method for generating celeriac cybrids |
title_full_unstemmed | An asymmetric protoplast fusion and screening method for generating celeriac cybrids |
title_short | An asymmetric protoplast fusion and screening method for generating celeriac cybrids |
title_sort | asymmetric protoplast fusion and screening method for generating celeriac cybrids |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7907277/ https://www.ncbi.nlm.nih.gov/pubmed/33633203 http://dx.doi.org/10.1038/s41598-021-83970-y |
work_keys_str_mv | AT bruznicansilvia anasymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT eeckhauttom anasymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT vanhuylenbroeckjohan anasymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT dekeyserellen anasymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT declercqherve anasymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT geelendanny anasymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT bruznicansilvia asymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT eeckhauttom asymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT vanhuylenbroeckjohan asymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT dekeyserellen asymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT declercqherve asymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids AT geelendanny asymmetricprotoplastfusionandscreeningmethodforgeneratingceleriaccybrids |