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Protoplast-Based Transient Expression and Gene Editing in Shrub Willow (Salix purpurea L.)
Shrub willows (Salix section Vetrix) are grown as a bioenergy crop in multiple countries and as ornamentals across the northern hemisphere. To facilitate the breeding and genetic advancement of shrub willow, there is a strong interest in the characterization and functional validation of genes involv...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9784668/ https://www.ncbi.nlm.nih.gov/pubmed/36559601 http://dx.doi.org/10.3390/plants11243490 |
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author | Hyden, Brennan Yuan, Guoliang Liu, Yang Smart, Lawrence B. Tuskan, Gerald A. Yang, Xiaohan |
author_facet | Hyden, Brennan Yuan, Guoliang Liu, Yang Smart, Lawrence B. Tuskan, Gerald A. Yang, Xiaohan |
author_sort | Hyden, Brennan |
collection | PubMed |
description | Shrub willows (Salix section Vetrix) are grown as a bioenergy crop in multiple countries and as ornamentals across the northern hemisphere. To facilitate the breeding and genetic advancement of shrub willow, there is a strong interest in the characterization and functional validation of genes involved in plant growth and biomass production. While protocols for shoot regeneration in tissue culture and production of stably transformed lines have greatly advanced this research in the closely related genus Populus, a lack of efficient methods for regeneration and transformation has stymied similar advancements in willow functional genomics. Moreover, transient expression assays in willow have been limited to callus tissue and hairy root systems. Here we report an efficient method for protoplast isolation from S. purpurea leaf tissue, along with transient overexpression and CRISPR-Cas9 mediated mutations. This is the first such report of transient gene expression in Salix protoplasts as well as the first application of CRISPR technology in this genus. These new capabilities pave the way for future functional genomics studies in this important bioenergy and ornamental crop. |
format | Online Article Text |
id | pubmed-9784668 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-97846682022-12-24 Protoplast-Based Transient Expression and Gene Editing in Shrub Willow (Salix purpurea L.) Hyden, Brennan Yuan, Guoliang Liu, Yang Smart, Lawrence B. Tuskan, Gerald A. Yang, Xiaohan Plants (Basel) Brief Report Shrub willows (Salix section Vetrix) are grown as a bioenergy crop in multiple countries and as ornamentals across the northern hemisphere. To facilitate the breeding and genetic advancement of shrub willow, there is a strong interest in the characterization and functional validation of genes involved in plant growth and biomass production. While protocols for shoot regeneration in tissue culture and production of stably transformed lines have greatly advanced this research in the closely related genus Populus, a lack of efficient methods for regeneration and transformation has stymied similar advancements in willow functional genomics. Moreover, transient expression assays in willow have been limited to callus tissue and hairy root systems. Here we report an efficient method for protoplast isolation from S. purpurea leaf tissue, along with transient overexpression and CRISPR-Cas9 mediated mutations. This is the first such report of transient gene expression in Salix protoplasts as well as the first application of CRISPR technology in this genus. These new capabilities pave the way for future functional genomics studies in this important bioenergy and ornamental crop. MDPI 2022-12-13 /pmc/articles/PMC9784668/ /pubmed/36559601 http://dx.doi.org/10.3390/plants11243490 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Brief Report Hyden, Brennan Yuan, Guoliang Liu, Yang Smart, Lawrence B. Tuskan, Gerald A. Yang, Xiaohan Protoplast-Based Transient Expression and Gene Editing in Shrub Willow (Salix purpurea L.) |
title | Protoplast-Based Transient Expression and Gene Editing in Shrub Willow (Salix purpurea L.) |
title_full | Protoplast-Based Transient Expression and Gene Editing in Shrub Willow (Salix purpurea L.) |
title_fullStr | Protoplast-Based Transient Expression and Gene Editing in Shrub Willow (Salix purpurea L.) |
title_full_unstemmed | Protoplast-Based Transient Expression and Gene Editing in Shrub Willow (Salix purpurea L.) |
title_short | Protoplast-Based Transient Expression and Gene Editing in Shrub Willow (Salix purpurea L.) |
title_sort | protoplast-based transient expression and gene editing in shrub willow (salix purpurea l.) |
topic | Brief Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9784668/ https://www.ncbi.nlm.nih.gov/pubmed/36559601 http://dx.doi.org/10.3390/plants11243490 |
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