Stepwise artificial evolution of an Sw-5b immune receptor extends its resistance spectrum against resistance-breaking isolates of Tomato spotted wilt virus

被引:21
作者
Huang, Haining [1 ,2 ]
Huang, Shen [1 ,2 ]
Li, Jia [1 ,2 ]
Wang, Huiyuan [1 ,2 ]
Zhao, Yaqian [1 ,2 ]
Feng, Mingfeng [1 ,2 ]
Dai, Jing [1 ,2 ]
Wang, Tongkai [1 ,2 ]
Zhu, Min [1 ,2 ]
Tao, Xiaorong [1 ,2 ]
机构
[1] Nanjing Agr Univ, Dept Plant Pathol, Nanjing, Peoples R China
[2] Nanjing Agr Univ, Key Lab Plant Immun, Nanjing, Peoples R China
基金
中国国家自然科学基金;
关键词
NLR; immune receptor; artificial evolution; Sw-5b; Tomato spotted wilt virus; resistance breaking; MOVEMENT PROTEIN NSM; HYPERSENSITIVE RESPONSE; NUCLEOTIDE-BINDING; PLANT NLRS; 1ST REPORT; GENE; DOMAIN; TOSPOVIRUSES; SPECIFICITY; RECOGNITION;
D O I
10.1111/pbi.13641
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Plants use intracellular nucleotide-binding leucine-rich repeat immune receptors (NLRs) to recognize pathogen-encoded effectors and initiate immune responses. Tomato spotted wilt virus (TSWV), which has been found to infect >1000 plant species, is among the most destructive plant viruses worldwide. The Sw-5b is the most effective and widely used resistance gene in tomato breeding to control TSWV. However, broad application of tomato cultivars carrying Sw-5b has resulted in an emergence of resistance-breaking (RB) TSWV. Therefore, new effective genes are urgently needed to prevent further RB TSWV outbreaks. In this study, we conducted artificial evolution to select Sw-5b mutants that could extend the resistance spectrum against TSWV RB isolates. Unlike regular NLRs, Sw-5b detects viral elicitor NSm using both the N-terminal Solanaceae-specific domain (SD) and the C-terminal LRR domain in a two-step recognition process. Our attempts to select gain-of-function mutants by random mutagenesis involving either the SD or the LRR of Sw-5b failed; therefore, we adopted a stepwise strategy, first introducing a NSmRB-responsive mutation at the R927 residue in the LRR, followed by random mutagenesis involving the Sw-5b SD domain. Using this strategy, we obtained Sw-5b(L33P/K319E/R927A) and Sw-5b(L33P/K319E/R927Q) mutants, which are effective against TSWV RB carrying the NSmC118Y or NSmT120N mutation, and against other American-type tospoviruses. Thus, we were able to extend the resistance spectrum of Sw-5b; the selected Sw-5b mutants will provide new gene resources to control RB TSWV.
引用
收藏
页码:2164 / 2176
页数:13
相关论文
共 67 条
[31]   A Plant Immune Receptor Adopts a Two-Step Recognition Mechanism to Enhance Viral Effector Perception [J].
Li, Jia ;
Huang, Haining ;
Zhu, Min ;
Huang, Shen ;
Zhang, Wenhua ;
Dinesh-Kumar, Savithramma P. ;
Tao, Xiaorong .
MOLECULAR PLANT, 2019, 12 (02) :248-262
[32]   Evolutionary analysis of tomato Sw-5 resistance-breaking isolates of Tomato spotted wilt virus [J].
Lopez, Carmelo ;
Aramburu, Jose ;
Galipienso, Luis ;
Soler, Salvador ;
Nuez, Fernando ;
Rubio, Luis .
JOURNAL OF GENERAL VIROLOGY, 2011, 92 :210-215
[33]   Dual Regulatory Roles of the Extended N Terminus for Activation of the Tomato Mi-1.2 Resistance Protein [J].
Lukasik-Shreepaathy, Ewa ;
Slootweg, Erik ;
Richter, Hanna ;
Goverse, Aska ;
Cornelissen, Ben J. C. ;
Takken, Frank L. W. .
MOLECULAR PLANT-MICROBE INTERACTIONS, 2012, 25 (08) :1045-1057
[34]   Direct pathogen-induced assembly of an NLR immune receptor complex to form a holoenzyme [J].
Ma, Shoucai ;
Lapin, Dmitry ;
Liu, Li ;
Sun, Yue ;
Song, Wen ;
Zhang, Xiaoxiao ;
Logemann, Elke ;
Yu, Dongli ;
Wang, Jia ;
Jirschitzka, Jan ;
Han, Zhifu ;
Schulze-Lefert, Paul ;
Parker, Jane E. ;
Chai, Jijie .
SCIENCE, 2020, 370 (6521) :1184-+
[35]   A paralogous decoy protects Phytophthora sojae apoplastic effector PsXEG1 from a host inhibitor [J].
Ma, Zhenchuan ;
Zhu, Lin ;
Song, Tianqiao ;
Wang, Yang ;
Zhang, Qi ;
Xia, Yeqiang ;
Qiu, Min ;
Lin, Yachun ;
Li, Haiyang ;
Kong, Liang ;
Fang, Yufeng ;
Ye, Wenwu ;
Wang, Yan ;
Dong, Suomeng ;
Zheng, Xiaobo ;
Tyler, Brett M. ;
Wang, Yuanchao .
SCIENCE, 2017, 355 (6326) :710-714
[36]   A Phytophthora sojae Glycoside Hydrolase 12 Protein Is a Major Virulence Factor during Soybean Infection and Is Recognized as a PAMP [J].
Ma, Zhenchuan ;
Song, Tianqiao ;
Zhu, Lin ;
Ye, Wenwu ;
Wang, Yang ;
Shao, Yuanyuan ;
Dong, Suomeng ;
Zhang, Zhengguang ;
Dou, Daolong ;
Zheng, Xiaobo ;
Tyler, Brett M. ;
Wang, Yuanchao .
PLANT CELL, 2015, 27 (07) :2057-2072
[37]   NLR functions in plant and animal immune systems: so far and yet so close [J].
Maekawa, Takaki ;
Kufer, Thomas A. ;
Schulze-Lefert, Paul .
NATURE IMMUNOLOGY, 2011, 12 (09) :818-826
[38]   Structure of the activated ROQ1 resistosome directly recognizing the pathogen effector XopQ [J].
Martin, Raoul ;
Qi, Tiancong ;
Zhang, Haibo ;
Liu, Furong ;
King, Miles ;
Toth, Claire ;
Nogales, Eva ;
Staskawicz, Brian J. .
SCIENCE, 2020, 370 (6521) :1185-+
[39]   Perspectives on intracellular perception of plant viruses [J].
Meier, Nathan ;
Hatch, Cameron ;
Nagalakshmi, Ugrappa ;
Dinesh-Kumar, Savithramma P. .
MOLECULAR PLANT PATHOLOGY, 2019, 20 (09) :1185-1190
[40]   Genome-wide analysis of NBS-LRR-encoding genes in Arabidopsis [J].
Meyers, BC ;
Kozik, A ;
Griego, A ;
Kuang, HH ;
Michelmore, RW .
PLANT CELL, 2003, 15 (04) :809-834