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CRISPR/Cas9-Mediated Multiplex Genome Editing of JAGGED Gene in Brassica napus L.

Pod shattering resistance is an essential component to achieving a high yield, which is a substantial objective in polyploid rapeseed cultivation. Previous studies have suggested that the Arabidopsis JAGGED (JAG) gene is a key factor implicated in the regulatory web of dehiscence fruit. However, its...

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Autores principales: Zaman, Qamar U, Chu, Wen, Hao, Mengyu, Shi, Yuqin, Sun, Mengdan, Sang, Shi-Fei, Mei, Desheng, Cheng, Hongtao, Liu, Jia, Li, Chao, Hu, Qiong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6921047/
https://www.ncbi.nlm.nih.gov/pubmed/31726660
http://dx.doi.org/10.3390/biom9110725
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author Zaman, Qamar U
Chu, Wen
Hao, Mengyu
Shi, Yuqin
Sun, Mengdan
Sang, Shi-Fei
Mei, Desheng
Cheng, Hongtao
Liu, Jia
Li, Chao
Hu, Qiong
author_facet Zaman, Qamar U
Chu, Wen
Hao, Mengyu
Shi, Yuqin
Sun, Mengdan
Sang, Shi-Fei
Mei, Desheng
Cheng, Hongtao
Liu, Jia
Li, Chao
Hu, Qiong
author_sort Zaman, Qamar U
collection PubMed
description Pod shattering resistance is an essential component to achieving a high yield, which is a substantial objective in polyploid rapeseed cultivation. Previous studies have suggested that the Arabidopsis JAGGED (JAG) gene is a key factor implicated in the regulatory web of dehiscence fruit. However, its role in controlling pod shattering resistance in oilseed rape is still unknown. In this study, multiplex genome editing was carried out by the CRISPR/Cas9 system on five homoeologs (BnJAG.A02, BnJAG.C02, BnJAG.C06, BnJAG.A07, and BnJAG.A08) of the JAG gene. Knockout mutagenesis of all homoeologs drastically affected the development of the lateral organs in organizing pod shape and size. The cylindrical body of the pod comprised a number of undifferentiated cells like a callus, without distinctive valves, replum, septum, and valve margins. Pseudoseeds were produced, which were divided into two halves with an incomplete layer of cells (probably septum) that separated the undifferentiated cells. These mutants were not capable of generating any productive seeds for further generations. However, one mutant line was identified in which only a BnJAG.A08-NUB-Like paralog of the JAG gene was mutated. Knockout mutagenesis in BnJAG.A08-NUB gene caused significant changes in the pod dehiscence zone. The replum region of the mutant was increased to a great extent, resulting in enlarged cell size, bumpy fruit, and reduced length compared with the wild type. A higher replum–valve joint area may have increased the resistance to pod shattering by ~2-fold in JAG mutants compared with wild type. Our results offer a basis for understanding variations in Brassica napus fruit by mutating JAG genes and providing a way forward for other Brassicaceae species.
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spelling pubmed-69210472019-12-24 CRISPR/Cas9-Mediated Multiplex Genome Editing of JAGGED Gene in Brassica napus L. Zaman, Qamar U Chu, Wen Hao, Mengyu Shi, Yuqin Sun, Mengdan Sang, Shi-Fei Mei, Desheng Cheng, Hongtao Liu, Jia Li, Chao Hu, Qiong Biomolecules Article Pod shattering resistance is an essential component to achieving a high yield, which is a substantial objective in polyploid rapeseed cultivation. Previous studies have suggested that the Arabidopsis JAGGED (JAG) gene is a key factor implicated in the regulatory web of dehiscence fruit. However, its role in controlling pod shattering resistance in oilseed rape is still unknown. In this study, multiplex genome editing was carried out by the CRISPR/Cas9 system on five homoeologs (BnJAG.A02, BnJAG.C02, BnJAG.C06, BnJAG.A07, and BnJAG.A08) of the JAG gene. Knockout mutagenesis of all homoeologs drastically affected the development of the lateral organs in organizing pod shape and size. The cylindrical body of the pod comprised a number of undifferentiated cells like a callus, without distinctive valves, replum, septum, and valve margins. Pseudoseeds were produced, which were divided into two halves with an incomplete layer of cells (probably septum) that separated the undifferentiated cells. These mutants were not capable of generating any productive seeds for further generations. However, one mutant line was identified in which only a BnJAG.A08-NUB-Like paralog of the JAG gene was mutated. Knockout mutagenesis in BnJAG.A08-NUB gene caused significant changes in the pod dehiscence zone. The replum region of the mutant was increased to a great extent, resulting in enlarged cell size, bumpy fruit, and reduced length compared with the wild type. A higher replum–valve joint area may have increased the resistance to pod shattering by ~2-fold in JAG mutants compared with wild type. Our results offer a basis for understanding variations in Brassica napus fruit by mutating JAG genes and providing a way forward for other Brassicaceae species. MDPI 2019-11-12 /pmc/articles/PMC6921047/ /pubmed/31726660 http://dx.doi.org/10.3390/biom9110725 Text en © 2019 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zaman, Qamar U
Chu, Wen
Hao, Mengyu
Shi, Yuqin
Sun, Mengdan
Sang, Shi-Fei
Mei, Desheng
Cheng, Hongtao
Liu, Jia
Li, Chao
Hu, Qiong
CRISPR/Cas9-Mediated Multiplex Genome Editing of JAGGED Gene in Brassica napus L.
title CRISPR/Cas9-Mediated Multiplex Genome Editing of JAGGED Gene in Brassica napus L.
title_full CRISPR/Cas9-Mediated Multiplex Genome Editing of JAGGED Gene in Brassica napus L.
title_fullStr CRISPR/Cas9-Mediated Multiplex Genome Editing of JAGGED Gene in Brassica napus L.
title_full_unstemmed CRISPR/Cas9-Mediated Multiplex Genome Editing of JAGGED Gene in Brassica napus L.
title_short CRISPR/Cas9-Mediated Multiplex Genome Editing of JAGGED Gene in Brassica napus L.
title_sort crispr/cas9-mediated multiplex genome editing of jagged gene in brassica napus l.
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6921047/
https://www.ncbi.nlm.nih.gov/pubmed/31726660
http://dx.doi.org/10.3390/biom9110725
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