Cargando…
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...
Autores principales: | , , , , , , , , , , , |
---|---|
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 |
_version_ | 1785103316734181376 |
---|---|
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. |
format | Online Article Text |
id | pubmed-10487758 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT leeyerin crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT kokiseong crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT leehyeeun crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT leeeunsu crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT hankoeun crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT yoojaeyong crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT vubichngoc crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT choihana crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT leeyoona crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT hongjongchan crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT leekyunoh crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress AT kimdosun crisprcas9mediatedhy5geneeditingreducesgrowthinhibitioninchinesecabbagebrassicarapaundererstress |