文献类型： 外文期刊

作者： Sun, Lin ¹ ; Alariqi, Muna ¹ ; Wang, Yaxin ¹ ; Wang, Qiongqiong ¹ ; Xu, Zhongping ¹ ; Zafar, Muhammad Naeem ¹ ; Yang, Guangqin ¹ ; Jia, Ruoyu ¹ ; Hussain, Amjad ¹ ; Chen, Yilin ¹ ; Ding, Xiao ¹ ; Zhou, Jiawei ¹ ; Wang, Guanying ¹ ; Wang, Fuqiu ¹ ; Li, Jianying ¹ ; Zou, Jiawei ¹ ; Zhu, Xiangqian ¹ ; Yu, Lu ¹ ; Sun, Yiwen ¹ ; Liang, Sijia ¹ ; Hui, Fengjiao ¹ ; Chen, Luo ¹ ; Guo, Weifeng ⁴ ; Wang, Yanqin ⁴ ; Zhu, Huaguo ⁵ ; Lindsey, Keith ⁶ ; Nie, Xinhui ⁷ ; Zhang, Xianlong ¹ ; Jin, Shuangxia ¹ ;

作者机构： 1.Huazhong Agr Univ, Hubei Hongshan Lab, Natl Key Lab Crop Genet Improvement, Wuhan 430070, Hubei, Peoples R China

2.Shandong Acad Agr Sci, Inst Ind Crops, Jinan 250100, Shandong, Peoples R China

3.Sanaa Univ, Dept Agron & Pastures, Fac Agr, Sanaa, Yemen

4.Tarim Univ, Xinjiang Prod & Construct Corps Key Lab Protect &, Alaer 843300, Xinjiang, Peoples R China

5.Huanggang Normal Univ, Coll Biol & Agr Resources, Huanggang 438000, Hubei, Peoples R China

6.Univ Durham, Dept Biosci, Durham DH1 3LE, England

7.Shihezi Univ, Agr Coll, Key Lab Oasis Ecol Agr Xinjiang Bingtuan, Shihezi, Xinjiang, Peoples R China

关键词： cotton; CRISPR/Cas9; GhEPS15; GhMLP423; high-throughput genome editing; plant-insect interaction

期刊名称：ADVANCED SCIENCE （ 影响因子：15.1； 五年影响因子：16.7 ）

ISSN：

年卷期： 2023 年

页码：

收录情况： SCI

摘要： 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 approximate to 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. A high-throughput screening strategy for cotton endogenous insect-resistance-related genes is established by a CRISPR/Cas9-mediated large-scale mutant library. It identifies promising genes related to plant innate insect-resistance, including major latex-like protein 423 (GhMLP423). GhMLP423 enhanced plant immunity to chewing and sap-sacking insects via the induction of GhEPS15 that initiates the systemic acquired resistance elicited by the systemically propagating waves of Ca2+-mediated ROS signaling.image