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Multiplexed CRISPR/Cas9-Mediated Knockout of Laccase Genes in Salvia miltiorrhiza Revealed Their Roles in Growth, Development, and Metabolism

Laccases are multicopper-containing glycoproteins related to monolignol oxidation and polymerization. These properties indicate that laccases may be involved in the formation of important medicinal phenolic acid compounds in Salvia miltiorrhiza such as salvianolic acid B (SAB), which is used for car...

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Autores principales: Zhou, Zheng, Li, Qing, Xiao, Liang, Wang, Yun, Feng, Jingxian, Bu, Qitao, Xiao, Ying, Hao, Kai, Guo, Meili, Chen, Wansheng, Zhang, Lei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8014014/
https://www.ncbi.nlm.nih.gov/pubmed/33815454
http://dx.doi.org/10.3389/fpls.2021.647768
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author Zhou, Zheng
Li, Qing
Xiao, Liang
Wang, Yun
Feng, Jingxian
Bu, Qitao
Xiao, Ying
Hao, Kai
Guo, Meili
Chen, Wansheng
Zhang, Lei
author_facet Zhou, Zheng
Li, Qing
Xiao, Liang
Wang, Yun
Feng, Jingxian
Bu, Qitao
Xiao, Ying
Hao, Kai
Guo, Meili
Chen, Wansheng
Zhang, Lei
author_sort Zhou, Zheng
collection PubMed
description Laccases are multicopper-containing glycoproteins related to monolignol oxidation and polymerization. These properties indicate that laccases may be involved in the formation of important medicinal phenolic acid compounds in Salvia miltiorrhiza such as salvianolic acid B (SAB), which is used for cardiovascular disease treatment. To date, 29 laccases have been found in S. miltiorrhiza (SmLACs), and some of which (SmLAC7 and SmLAC20) have been reported to influence the synthesis of phenolic acids. Because of the functional redundancy of laccase genes, their roles in S. miltiorrhiza are poorly understood. In this study, the CRISPR/Cas9 system was used for targeting conserved domains to knockout multiple genes of laccase family in S. miltiorrhiza. The expressions of target laccase genes as well as the phenolic acid biosynthesis key genes decrease dramatically in editing lines. Additionally, the growth and development of hairy roots was significantly retarded in the gene-edited lines. The cross-sections examination of laccase mutant hairy roots showed that the root development was abnormal and the xylem cells in the edited lines became larger and looser than those in the wild type. Additionally, the accumulation of RA as well as SAB was decreased, and the lignin content was nearly undetectable. It suggested that SmLACs play key roles in development and lignin formation in the root of S. miltiorrhiza and they are necessary for phenolic acids biosynthesis.
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spelling pubmed-80140142021-04-02 Multiplexed CRISPR/Cas9-Mediated Knockout of Laccase Genes in Salvia miltiorrhiza Revealed Their Roles in Growth, Development, and Metabolism Zhou, Zheng Li, Qing Xiao, Liang Wang, Yun Feng, Jingxian Bu, Qitao Xiao, Ying Hao, Kai Guo, Meili Chen, Wansheng Zhang, Lei Front Plant Sci Plant Science Laccases are multicopper-containing glycoproteins related to monolignol oxidation and polymerization. These properties indicate that laccases may be involved in the formation of important medicinal phenolic acid compounds in Salvia miltiorrhiza such as salvianolic acid B (SAB), which is used for cardiovascular disease treatment. To date, 29 laccases have been found in S. miltiorrhiza (SmLACs), and some of which (SmLAC7 and SmLAC20) have been reported to influence the synthesis of phenolic acids. Because of the functional redundancy of laccase genes, their roles in S. miltiorrhiza are poorly understood. In this study, the CRISPR/Cas9 system was used for targeting conserved domains to knockout multiple genes of laccase family in S. miltiorrhiza. The expressions of target laccase genes as well as the phenolic acid biosynthesis key genes decrease dramatically in editing lines. Additionally, the growth and development of hairy roots was significantly retarded in the gene-edited lines. The cross-sections examination of laccase mutant hairy roots showed that the root development was abnormal and the xylem cells in the edited lines became larger and looser than those in the wild type. Additionally, the accumulation of RA as well as SAB was decreased, and the lignin content was nearly undetectable. It suggested that SmLACs play key roles in development and lignin formation in the root of S. miltiorrhiza and they are necessary for phenolic acids biosynthesis. Frontiers Media S.A. 2021-03-18 /pmc/articles/PMC8014014/ /pubmed/33815454 http://dx.doi.org/10.3389/fpls.2021.647768 Text en Copyright © 2021 Zhou, Li, Xiao, Wang, Feng, Bu, Xiao, Hao, Guo, Chen and Zhang. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Zhou, Zheng
Li, Qing
Xiao, Liang
Wang, Yun
Feng, Jingxian
Bu, Qitao
Xiao, Ying
Hao, Kai
Guo, Meili
Chen, Wansheng
Zhang, Lei
Multiplexed CRISPR/Cas9-Mediated Knockout of Laccase Genes in Salvia miltiorrhiza Revealed Their Roles in Growth, Development, and Metabolism
title Multiplexed CRISPR/Cas9-Mediated Knockout of Laccase Genes in Salvia miltiorrhiza Revealed Their Roles in Growth, Development, and Metabolism
title_full Multiplexed CRISPR/Cas9-Mediated Knockout of Laccase Genes in Salvia miltiorrhiza Revealed Their Roles in Growth, Development, and Metabolism
title_fullStr Multiplexed CRISPR/Cas9-Mediated Knockout of Laccase Genes in Salvia miltiorrhiza Revealed Their Roles in Growth, Development, and Metabolism
title_full_unstemmed Multiplexed CRISPR/Cas9-Mediated Knockout of Laccase Genes in Salvia miltiorrhiza Revealed Their Roles in Growth, Development, and Metabolism
title_short Multiplexed CRISPR/Cas9-Mediated Knockout of Laccase Genes in Salvia miltiorrhiza Revealed Their Roles in Growth, Development, and Metabolism
title_sort multiplexed crispr/cas9-mediated knockout of laccase genes in salvia miltiorrhiza revealed their roles in growth, development, and metabolism
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8014014/
https://www.ncbi.nlm.nih.gov/pubmed/33815454
http://dx.doi.org/10.3389/fpls.2021.647768
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