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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...

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Autores principales: Hyden, Brennan, Yuan, Guoliang, Liu, Yang, Smart, Lawrence B., Tuskan, Gerald A., Yang, Xiaohan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
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.
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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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