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CRISPR/Cas9-Mediated HY5 Gene Editing Reduces Growth Inhibition in Chinese Cabbage (Brassica rapa) under ER Stress

Various stresses can affect the quality and yield of crops, including vegetables. In this study, CRISPR/Cas9 technology was employed to examine the role of the ELONGATED HYPOCOTYL 5 (HY5) gene in influencing the growth of Chinese cabbage (Brassica rapa). Single guide RNAs (sgRNAs) were designed to t...

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Autores principales: Lee, Ye Rin, Ko, Ki Seong, Lee, Hye Eun, Lee, Eun Su, Han, Koeun, Yoo, Jae Yong, Vu, Bich Ngoc, Choi, Ha Na, Lee, Yoo Na, Hong, Jong Chan, Lee, Kyun Oh, Kim, Do Sun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10487758/
https://www.ncbi.nlm.nih.gov/pubmed/37685921
http://dx.doi.org/10.3390/ijms241713105
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author Lee, Ye Rin
Ko, Ki Seong
Lee, Hye Eun
Lee, Eun Su
Han, Koeun
Yoo, Jae Yong
Vu, Bich Ngoc
Choi, Ha Na
Lee, Yoo Na
Hong, Jong Chan
Lee, Kyun Oh
Kim, Do Sun
author_facet Lee, Ye Rin
Ko, Ki Seong
Lee, Hye Eun
Lee, Eun Su
Han, Koeun
Yoo, Jae Yong
Vu, Bich Ngoc
Choi, Ha Na
Lee, Yoo Na
Hong, Jong Chan
Lee, Kyun Oh
Kim, Do Sun
author_sort Lee, Ye Rin
collection PubMed
description Various stresses can affect the quality and yield of crops, including vegetables. In this study, CRISPR/Cas9 technology was employed to examine the role of the ELONGATED HYPOCOTYL 5 (HY5) gene in influencing the growth of Chinese cabbage (Brassica rapa). Single guide RNAs (sgRNAs) were designed to target the HY5 gene, and deep-sequencing analysis confirmed the induction of mutations in the bZIP domain of the gene. To investigate the response of Chinese cabbage to endoplasmic reticulum (ER) stress, plants were treated with tunicamycin (TM). Both wild-type and hy5 mutant plants showed increased growth inhibition with increasing TM concentration. However, the hy5 mutant plants displayed less severe growth inhibition compared to the wild type. Using nitroblue tetrazolium (NBT) and 3,3′-diaminobenzidine (DAB) staining methods, we determined the amount of reactive oxygen species (ROS) produced under ER stress conditions, and found that the hy5 mutant plants generated lower levels of ROS compared to the wild type. Under ER stress conditions, the hy5 mutant plants exhibited lower expression levels of UPR- and cell death-related genes than the wild type. These results indicate that CRISPR/Cas9-mediated editing of the HY5 gene can mitigate growth inhibition in Chinese cabbage under stresses, improving the quality and yield of crops.
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spelling pubmed-104877582023-09-09 CRISPR/Cas9-Mediated HY5 Gene Editing Reduces Growth Inhibition in Chinese Cabbage (Brassica rapa) under ER Stress Lee, Ye Rin Ko, Ki Seong Lee, Hye Eun Lee, Eun Su Han, Koeun Yoo, Jae Yong Vu, Bich Ngoc Choi, Ha Na Lee, Yoo Na Hong, Jong Chan Lee, Kyun Oh Kim, Do Sun Int J Mol Sci Article Various stresses can affect the quality and yield of crops, including vegetables. In this study, CRISPR/Cas9 technology was employed to examine the role of the ELONGATED HYPOCOTYL 5 (HY5) gene in influencing the growth of Chinese cabbage (Brassica rapa). Single guide RNAs (sgRNAs) were designed to target the HY5 gene, and deep-sequencing analysis confirmed the induction of mutations in the bZIP domain of the gene. To investigate the response of Chinese cabbage to endoplasmic reticulum (ER) stress, plants were treated with tunicamycin (TM). Both wild-type and hy5 mutant plants showed increased growth inhibition with increasing TM concentration. However, the hy5 mutant plants displayed less severe growth inhibition compared to the wild type. Using nitroblue tetrazolium (NBT) and 3,3′-diaminobenzidine (DAB) staining methods, we determined the amount of reactive oxygen species (ROS) produced under ER stress conditions, and found that the hy5 mutant plants generated lower levels of ROS compared to the wild type. Under ER stress conditions, the hy5 mutant plants exhibited lower expression levels of UPR- and cell death-related genes than the wild type. These results indicate that CRISPR/Cas9-mediated editing of the HY5 gene can mitigate growth inhibition in Chinese cabbage under stresses, improving the quality and yield of crops. MDPI 2023-08-23 /pmc/articles/PMC10487758/ /pubmed/37685921 http://dx.doi.org/10.3390/ijms241713105 Text en © 2023 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 Article
Lee, Ye Rin
Ko, Ki Seong
Lee, Hye Eun
Lee, Eun Su
Han, Koeun
Yoo, Jae Yong
Vu, Bich Ngoc
Choi, Ha Na
Lee, Yoo Na
Hong, Jong Chan
Lee, Kyun Oh
Kim, Do Sun
CRISPR/Cas9-Mediated HY5 Gene Editing Reduces Growth Inhibition in Chinese Cabbage (Brassica rapa) under ER Stress
title CRISPR/Cas9-Mediated HY5 Gene Editing Reduces Growth Inhibition in Chinese Cabbage (Brassica rapa) under ER Stress
title_full CRISPR/Cas9-Mediated HY5 Gene Editing Reduces Growth Inhibition in Chinese Cabbage (Brassica rapa) under ER Stress
title_fullStr CRISPR/Cas9-Mediated HY5 Gene Editing Reduces Growth Inhibition in Chinese Cabbage (Brassica rapa) under ER Stress
title_full_unstemmed CRISPR/Cas9-Mediated HY5 Gene Editing Reduces Growth Inhibition in Chinese Cabbage (Brassica rapa) under ER Stress
title_short CRISPR/Cas9-Mediated HY5 Gene Editing Reduces Growth Inhibition in Chinese Cabbage (Brassica rapa) under ER Stress
title_sort crispr/cas9-mediated hy5 gene editing reduces growth inhibition in chinese cabbage (brassica rapa) under er stress
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10487758/
https://www.ncbi.nlm.nih.gov/pubmed/37685921
http://dx.doi.org/10.3390/ijms241713105
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