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Virus‐induced gene silencing as a tool for functional studies in Cleome violacea
PREMISE: Cleomaceae is emerging as a promising family to investigate a wide range of phenomena, such as C(4) photosynthesis and floral diversity. However, functional techniques are lacking for elucidating this diversity. Herein, we establish virus‐induced gene silencing (VIGS) as a method of generat...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8202831/ https://www.ncbi.nlm.nih.gov/pubmed/34141499 http://dx.doi.org/10.1002/aps3.11435 |
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author | Carey, Shane Higuera‐Díaz, Mónica Mankowski, Peter Rocca, Alexandra Hall, Jocelyn C. |
author_facet | Carey, Shane Higuera‐Díaz, Mónica Mankowski, Peter Rocca, Alexandra Hall, Jocelyn C. |
author_sort | Carey, Shane |
collection | PubMed |
description | PREMISE: Cleomaceae is emerging as a promising family to investigate a wide range of phenomena, such as C(4) photosynthesis and floral diversity. However, functional techniques are lacking for elucidating this diversity. Herein, we establish virus‐induced gene silencing (VIGS) as a method of generating functional data for Cleome violacea, bolstering Cleomaceae as a model system. METHODS: We leveraged the sister relationship of Cleomaceae and Brassicaceae by using constructs readily available for Arabidopsis thaliana to provide initial information about the feasibility of VIGS in C. violacea. We then developed endogenous constructs to optimize VIGS efficiency and viability for fruit development. RESULTS: PHYTOENE DESATURASE was successfully downregulated in C. violacea using both heterologous and endogenous constructs. The endogenous construct had the highest degree of downregulation, with many plants displaying strong photobleaching. FRUITFULL‐treated plants were also successfully downregulated, with a high rate of survival but less effective silencing; only a small percentage of survivors showed a strong phenotype. DISCUSSION: Our optimized VIGS protocol in C. violacea enables functional gene analyses at different developmental stages. Additionally, C. violacea is amenable to heterologous knockdown, which suggests that a first pass using non‐endogenous constructs is a possible route to test additional species of Cleomaceae. |
format | Online Article Text |
id | pubmed-8202831 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-82028312021-06-16 Virus‐induced gene silencing as a tool for functional studies in Cleome violacea Carey, Shane Higuera‐Díaz, Mónica Mankowski, Peter Rocca, Alexandra Hall, Jocelyn C. Appl Plant Sci Application Article PREMISE: Cleomaceae is emerging as a promising family to investigate a wide range of phenomena, such as C(4) photosynthesis and floral diversity. However, functional techniques are lacking for elucidating this diversity. Herein, we establish virus‐induced gene silencing (VIGS) as a method of generating functional data for Cleome violacea, bolstering Cleomaceae as a model system. METHODS: We leveraged the sister relationship of Cleomaceae and Brassicaceae by using constructs readily available for Arabidopsis thaliana to provide initial information about the feasibility of VIGS in C. violacea. We then developed endogenous constructs to optimize VIGS efficiency and viability for fruit development. RESULTS: PHYTOENE DESATURASE was successfully downregulated in C. violacea using both heterologous and endogenous constructs. The endogenous construct had the highest degree of downregulation, with many plants displaying strong photobleaching. FRUITFULL‐treated plants were also successfully downregulated, with a high rate of survival but less effective silencing; only a small percentage of survivors showed a strong phenotype. DISCUSSION: Our optimized VIGS protocol in C. violacea enables functional gene analyses at different developmental stages. Additionally, C. violacea is amenable to heterologous knockdown, which suggests that a first pass using non‐endogenous constructs is a possible route to test additional species of Cleomaceae. John Wiley and Sons Inc. 2021-06-14 /pmc/articles/PMC8202831/ /pubmed/34141499 http://dx.doi.org/10.1002/aps3.11435 Text en © 2021 The Authors. Applications in Plant Sciences published by Wiley Periodicals LLC on behalf of Botanical Society of America. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Application Article Carey, Shane Higuera‐Díaz, Mónica Mankowski, Peter Rocca, Alexandra Hall, Jocelyn C. Virus‐induced gene silencing as a tool for functional studies in Cleome violacea |
title | Virus‐induced gene silencing as a tool for functional studies in Cleome violacea
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title_full | Virus‐induced gene silencing as a tool for functional studies in Cleome violacea
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title_fullStr | Virus‐induced gene silencing as a tool for functional studies in Cleome violacea
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title_full_unstemmed | Virus‐induced gene silencing as a tool for functional studies in Cleome violacea
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title_short | Virus‐induced gene silencing as a tool for functional studies in Cleome violacea
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title_sort | virus‐induced gene silencing as a tool for functional studies in cleome violacea |
topic | Application Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8202831/ https://www.ncbi.nlm.nih.gov/pubmed/34141499 http://dx.doi.org/10.1002/aps3.11435 |
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