Construction of Host Plant Insect‐Resistance Mutant Library by High‐Throughput CRISPR/Cas9 System and Identification of A Broad‐Spectrum Insect Resistance Gene
Abstract Insects pose significant challenges in cotton‐producing regions. Here, they describe a high‐throughput CRISPR/Cas9‐mediated large‐scale mutagenesis library targeting endogenous insect‐resistance‐related genes in cotton. This library targeted 502 previously identified genes using 968 sgRNAs,...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2024-01-01
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| Series: | Advanced Science |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/advs.202306157 |
| _version_ | 1827202379610587136 |
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| author | Lin Sun Muna Alariqi Yaxin Wang Qiongqiong Wang Zhongping Xu Muhammad Naeem Zafar Guangqin Yang Ruoyu Jia Amjad Hussain Yilin Chen Xiao Ding Jiawei Zhou Guanying Wang Fuqiu Wang Jianying Li Jiawei Zou Xiangqian Zhu Lu Yu Yiwen Sun Sijia Liang Fengjiao Hui Luo Chen Weifeng Guo Yanqin Wang Huaguo Zhu Keith Lindsey Xinhui Nie Xianlong Zhang Shuangxia Jin |
| author_facet | Lin Sun Muna Alariqi Yaxin Wang Qiongqiong Wang Zhongping Xu Muhammad Naeem Zafar Guangqin Yang Ruoyu Jia Amjad Hussain Yilin Chen Xiao Ding Jiawei Zhou Guanying Wang Fuqiu Wang Jianying Li Jiawei Zou Xiangqian Zhu Lu Yu Yiwen Sun Sijia Liang Fengjiao Hui Luo Chen Weifeng Guo Yanqin Wang Huaguo Zhu Keith Lindsey Xinhui Nie Xianlong Zhang Shuangxia Jin |
| author_sort | Lin Sun |
| collection | DOAJ |
| description | Abstract Insects pose significant challenges in cotton‐producing regions. Here, they describe a high‐throughput CRISPR/Cas9‐mediated large‐scale mutagenesis library targeting endogenous insect‐resistance‐related genes in cotton. This library targeted 502 previously identified genes using 968 sgRNAs, generated ≈2000 T0 plants and achieved 97.29% genome editing with efficient heredity, reaching upto 84.78%. Several potential resistance‐related mutants (10% of 200 lines) their identified that may contribute to cotton‐insect molecular interaction. Among these, they selected 139 and 144 lines showing decreased resistance to pest infestation and targeting major latex‐like protein 423 (GhMLP423) for in‐depth study. Overexpression of GhMLP423 enhanced insect resistance by activating the plant systemic acquired resistance (SAR) of salicylic acid (SA) and pathogenesis‐related (PR) genes. This activation is induced by an elevation of cytosolic calcium [Ca2+]cyt flux eliciting reactive oxygen species (ROS), which their demoted in GhMLP423 knockout (CR) plants. Protein‐protein interaction assays revealed that GhMLP423 interacted with a human epidermal growth factor receptor substrate15 (EPS15) protein at the cell membrane. Together, they regulated the systemically propagating waves of Ca2+ and ROS, which in turn induced SAR. Collectively, this large‐scale mutagenesis library provides an efficient strategy for functional genomics research of polyploid plant species and serves as a solid platform for genetic engineering of insect resistance. |
| first_indexed | 2024-03-08T11:21:01Z |
| format | Article |
| id | doaj.art-05fad6c545704c35a2e9550b7e7597be |
| institution | Directory Open Access Journal |
| issn | 2198-3844 |
| language | English |
| last_indexed | 2025-03-21T11:19:40Z |
| publishDate | 2024-01-01 |
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| series | Advanced Science |
| spelling | doaj.art-05fad6c545704c35a2e9550b7e7597be2024-07-02T09:51:26ZengWileyAdvanced Science2198-38442024-01-01114n/an/a10.1002/advs.202306157Construction of Host Plant Insect‐Resistance Mutant Library by High‐Throughput CRISPR/Cas9 System and Identification of A Broad‐Spectrum Insect Resistance GeneLin Sun0Muna Alariqi1Yaxin Wang2Qiongqiong Wang3Zhongping Xu4Muhammad Naeem Zafar5Guangqin Yang6Ruoyu Jia7Amjad Hussain8Yilin Chen9Xiao Ding10Jiawei Zhou11Guanying Wang12Fuqiu Wang13Jianying Li14Jiawei Zou15Xiangqian Zhu16Lu Yu17Yiwen Sun18Sijia Liang19Fengjiao Hui20Luo Chen21Weifeng Guo22Yanqin Wang23Huaguo Zhu24Keith Lindsey25Xinhui Nie26Xianlong Zhang27Shuangxia Jin28Hubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaXinjiang Production and Construction Corps Key Laboratory of Protection and Utilization of Biological Resources in Tarim Basin Tarim University Alaer Xinjiang 843300 ChinaXinjiang Production and Construction Corps Key Laboratory of Protection and Utilization of Biological Resources in Tarim Basin Tarim University Alaer Xinjiang 843300 ChinaCollege of Biology and Agricultural Resources Huanggang Normal University Huanggang Hubei 438000 ChinaDepartment of Biosciences Durham University Durham DH1 3LE UKKey Laboratory of Oasis Ecology Agricultural of Xinjiang Bingtuan Agricultural College Shihezi University Shihezi Xinjiang ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan Hubei 430070 P. R. ChinaAbstract Insects pose significant challenges in cotton‐producing regions. Here, they describe a high‐throughput CRISPR/Cas9‐mediated large‐scale mutagenesis library targeting endogenous insect‐resistance‐related genes in cotton. This library targeted 502 previously identified genes using 968 sgRNAs, generated ≈2000 T0 plants and achieved 97.29% genome editing with efficient heredity, reaching upto 84.78%. Several potential resistance‐related mutants (10% of 200 lines) their identified that may contribute to cotton‐insect molecular interaction. Among these, they selected 139 and 144 lines showing decreased resistance to pest infestation and targeting major latex‐like protein 423 (GhMLP423) for in‐depth study. Overexpression of GhMLP423 enhanced insect resistance by activating the plant systemic acquired resistance (SAR) of salicylic acid (SA) and pathogenesis‐related (PR) genes. This activation is induced by an elevation of cytosolic calcium [Ca2+]cyt flux eliciting reactive oxygen species (ROS), which their demoted in GhMLP423 knockout (CR) plants. Protein‐protein interaction assays revealed that GhMLP423 interacted with a human epidermal growth factor receptor substrate15 (EPS15) protein at the cell membrane. Together, they regulated the systemically propagating waves of Ca2+ and ROS, which in turn induced SAR. Collectively, this large‐scale mutagenesis library provides an efficient strategy for functional genomics research of polyploid plant species and serves as a solid platform for genetic engineering of insect resistance.https://doi.org/10.1002/advs.202306157cottonCRISPR/Cas9GhEPS15GhMLP423high‐throughput genome editingplant‐insect interaction |
| spellingShingle | Lin Sun Muna Alariqi Yaxin Wang Qiongqiong Wang Zhongping Xu Muhammad Naeem Zafar Guangqin Yang Ruoyu Jia Amjad Hussain Yilin Chen Xiao Ding Jiawei Zhou Guanying Wang Fuqiu Wang Jianying Li Jiawei Zou Xiangqian Zhu Lu Yu Yiwen Sun Sijia Liang Fengjiao Hui Luo Chen Weifeng Guo Yanqin Wang Huaguo Zhu Keith Lindsey Xinhui Nie Xianlong Zhang Shuangxia Jin Construction of Host Plant Insect‐Resistance Mutant Library by High‐Throughput CRISPR/Cas9 System and Identification of A Broad‐Spectrum Insect Resistance Gene Advanced Science cotton CRISPR/Cas9 GhEPS15 GhMLP423 high‐throughput genome editing plant‐insect interaction |
| title | Construction of Host Plant Insect‐Resistance Mutant Library by High‐Throughput CRISPR/Cas9 System and Identification of A Broad‐Spectrum Insect Resistance Gene |
| title_full | Construction of Host Plant Insect‐Resistance Mutant Library by High‐Throughput CRISPR/Cas9 System and Identification of A Broad‐Spectrum Insect Resistance Gene |
| title_fullStr | Construction of Host Plant Insect‐Resistance Mutant Library by High‐Throughput CRISPR/Cas9 System and Identification of A Broad‐Spectrum Insect Resistance Gene |
| title_full_unstemmed | Construction of Host Plant Insect‐Resistance Mutant Library by High‐Throughput CRISPR/Cas9 System and Identification of A Broad‐Spectrum Insect Resistance Gene |
| title_short | Construction of Host Plant Insect‐Resistance Mutant Library by High‐Throughput CRISPR/Cas9 System and Identification of A Broad‐Spectrum Insect Resistance Gene |
| title_sort | construction of host plant insect resistance mutant library by high throughput crispr cas9 system and identification of a broad spectrum insect resistance gene |
| topic | cotton CRISPR/Cas9 GhEPS15 GhMLP423 high‐throughput genome editing plant‐insect interaction |
| url | https://doi.org/10.1002/advs.202306157 |
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