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CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.)

Downy mildew of grapevine (Vitis vinifera L.), caused by the oomycete pathogen Plasmopara viticola, is one of the most serious concerns for grape production worldwide. It has been widely reported that the pathogenesis-related 4 (PR4) protein plays important roles in plant resistance to diseases. How...

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Autores principales: Li, Meng-Yuan, Jiao, Yun-Tong, Wang, Yu-Ting, Zhang, Na, Wang, Bian-Bian, Liu, Rui-Qi, Yin, Xiao, Xu, Yan, Liu, Guo-Tian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7458914/
https://www.ncbi.nlm.nih.gov/pubmed/32922821
http://dx.doi.org/10.1038/s41438-020-00371-4
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author Li, Meng-Yuan
Jiao, Yun-Tong
Wang, Yu-Ting
Zhang, Na
Wang, Bian-Bian
Liu, Rui-Qi
Yin, Xiao
Xu, Yan
Liu, Guo-Tian
author_facet Li, Meng-Yuan
Jiao, Yun-Tong
Wang, Yu-Ting
Zhang, Na
Wang, Bian-Bian
Liu, Rui-Qi
Yin, Xiao
Xu, Yan
Liu, Guo-Tian
author_sort Li, Meng-Yuan
collection PubMed
description Downy mildew of grapevine (Vitis vinifera L.), caused by the oomycete pathogen Plasmopara viticola, is one of the most serious concerns for grape production worldwide. It has been widely reported that the pathogenesis-related 4 (PR4) protein plays important roles in plant resistance to diseases. However, little is known about the role of PR4 in the defense of grapevine against P. viticola. In this study, we engineered loss-of-function mutations in the VvPR4b gene from the cultivar “Thompson Seedless” using the CRISPR/Cas9 system and evaluated the consequences for downy mildew resistance. Sequencing results showed that deletions were the main type of mutation introduced and that no off-target events occurred. Infection assays using leaf discs showed that, compared to wild-type plants, the VvPR4b knockout lines had increased susceptibility to P. viticola. This was accompanied by reduced accumulation of reactive oxygen species around stomata. Measurement of the relative genomic abundance of P. viticola in VvPR4b knockout lines also demonstrated that the mutants had increased susceptibility to the pathogen. Our results confirm that VvPR4b plays an active role in the defense of grapevine against downy mildew.
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spelling pubmed-74589142020-09-11 CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.) Li, Meng-Yuan Jiao, Yun-Tong Wang, Yu-Ting Zhang, Na Wang, Bian-Bian Liu, Rui-Qi Yin, Xiao Xu, Yan Liu, Guo-Tian Hortic Res Article Downy mildew of grapevine (Vitis vinifera L.), caused by the oomycete pathogen Plasmopara viticola, is one of the most serious concerns for grape production worldwide. It has been widely reported that the pathogenesis-related 4 (PR4) protein plays important roles in plant resistance to diseases. However, little is known about the role of PR4 in the defense of grapevine against P. viticola. In this study, we engineered loss-of-function mutations in the VvPR4b gene from the cultivar “Thompson Seedless” using the CRISPR/Cas9 system and evaluated the consequences for downy mildew resistance. Sequencing results showed that deletions were the main type of mutation introduced and that no off-target events occurred. Infection assays using leaf discs showed that, compared to wild-type plants, the VvPR4b knockout lines had increased susceptibility to P. viticola. This was accompanied by reduced accumulation of reactive oxygen species around stomata. Measurement of the relative genomic abundance of P. viticola in VvPR4b knockout lines also demonstrated that the mutants had increased susceptibility to the pathogen. Our results confirm that VvPR4b plays an active role in the defense of grapevine against downy mildew. Nature Publishing Group UK 2020-09-01 /pmc/articles/PMC7458914/ /pubmed/32922821 http://dx.doi.org/10.1038/s41438-020-00371-4 Text en © The Author(s) 2020 https://creativecommons.org/licenses/by/4.0/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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Li, Meng-Yuan
Jiao, Yun-Tong
Wang, Yu-Ting
Zhang, Na
Wang, Bian-Bian
Liu, Rui-Qi
Yin, Xiao
Xu, Yan
Liu, Guo-Tian
CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.)
title CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.)
title_full CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.)
title_fullStr CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.)
title_full_unstemmed CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.)
title_short CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.)
title_sort crispr/cas9-mediated vvpr4b editing decreases downy mildew resistance in grapevine (vitis vinifera l.)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7458914/
https://www.ncbi.nlm.nih.gov/pubmed/32922821
http://dx.doi.org/10.1038/s41438-020-00371-4
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