A robust high-throughput functional screening assay for plant pathogen effectors using the TMV-GFP vector
文献类型： 外文期刊
作者： Cao, Peng 1 ; Shi, Haotian 1 ; Zhang, Shuangxi 1 ; Chen, Jialan 2 ; Wang, Rongbo 3 ; Liu, Peiqing 3 ; Zhu, Yingfang 4 ; An, Yuyan 1 ; Zhang, Meixiang 1 ;
作者机构： 1.Shaanxi Normal Univ, Coll Life Sci, Xian 710119, Peoples R China
2.Nanjing Agr Univ, Dept Plant Pathol, Nanjing 210095, Peoples R China
3.Fujian Acad Agr Sci, Inst Plant Protect, Fujian Key Lab Monitoring & Integrated Management, Fuzhou 350003, Peoples R China
4.Henan Univ, Sch Life Sci, State Key Lab Crop Stress Adaptat & Improvement, Key Lab Cotton Biol, Kaifeng 475001, Peoples R China
关键词： TMV-GFP; effector; virulence; avirulence; functional screening; technical advance
期刊名称：PLANT JOURNAL （ 影响因子：7.2； 五年影响因子：7.9 ）
ISSN： 0960-7412
年卷期： 2024 年
页码：
收录情况： SCI
摘要： Uncovering the function of phytopathogen effectors is crucial for understanding mechanisms of pathogen pathogenicity and for improving our ability to protect plants from diseases. An increasing number of effectors have been predicted in various plant pathogens. Functional characterization of these effectors has become a major focus in the study of plant-pathogen interactions. In this study, we designed a novel screening system that combines the TMV (tobacco mosaic virus)-GFP vector and Agrobacterium-mediated transient expression in the model plant Nicotiana benthamiana. This system enables the rapid identification of effectors that interfere with plant immunity. The biological function of these effectors can be easily evaluated by observing the GFP fluorescence signal using a UV lamp within just a few days. To evaluate the TMV-GFP system, we initially tested it with well-described virulence and avirulence type III effectors from the bacterial pathogen Ralstonia solanacearum. After proving the accuracy and efficiency of the TMV-GFP system, we successfully screened a novel virulence effector, RipS1, using this approach. Furthermore, using the TMV-GFP system, we reproduced consistent results with previously known cytoplasmic effectors from a diverse array of pathogens. Additionally, we demonstrated the effectiveness of the TMV-GFP system in identifying apoplastic effectors. The easy operation, time-saving nature, broad effectiveness, and low technical requirements of the TMV-GFP system make it a promising approach for high-throughput screening of effectors with immune interference activity from various pathogens.
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