Lin Sun
Construction of Host Plant Insect‐Resistance Mutant Library by High‐Throughput CRISPR/Cas9 System and Identification of A Broad‐Spectrum Insect Resistance Gene
Sun, Lin; Alariqi, Muna; Wang, Yaxin; Wang, Qiongqiong; Xu, Zhongping; Zafar, Muhammad Naeem; Yang, Guangqin; Jia, Ruoyu; Hussain, Amjad; Chen, Yilin; Ding, Xiao; Zhou, Jiawei; Wang, Guanying; Wang, Fuqiu; Li, Jianying; Zou, Jiawei; Zhu, Xiangqian; Yu, Lu; Sun, Yiwen; Liang, Sijia; Hui, Fengjiao; Chen, Luo; Guo, Weifeng; Wang, Yanqin; Zhu, Huaguo; Lindsey, Keith; Nie, Xinhui; Zhang, Xianlong; Jin, Shuangxia
Authors
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
Professor Keith Lindsey keith.lindsey@durham.ac.uk
Professor
Xinhui Nie
Xianlong Zhang
Shuangxia Jin
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.
Citation
Sun, L., Alariqi, M., Wang, Y., Wang, Q., Xu, Z., Zafar, M. N., Yang, G., Jia, R., Hussain, A., Chen, Y., Ding, X., Zhou, J., Wang, G., Wang, F., Li, J., Zou, J., Zhu, X., Yu, L., Sun, Y., Liang, S., …Jin, S. (2024). 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, 11(4), Article 2306157. https://doi.org/10.1002/advs.202306157
Journal Article Type | Article |
---|---|
Acceptance Date | Oct 17, 2023 |
Online Publication Date | Nov 30, 2023 |
Publication Date | Jan 26, 2024 |
Deposit Date | Dec 4, 2023 |
Publicly Available Date | Dec 4, 2023 |
Journal | Advanced Science |
Electronic ISSN | 2198-3844 |
Publisher | Wiley Open Access |
Peer Reviewed | Peer Reviewed |
Volume | 11 |
Issue | 4 |
Article Number | 2306157 |
DOI | https://doi.org/10.1002/advs.202306157 |
Public URL | https://durham-repository.worktribe.com/output/1965518 |
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Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/
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