A robust high-throughput functional screening assay for plant pathogen effectors using the TMV-GFP vector.
Plant J
; 119(1): 617-631, 2024 Jul.
Article
in En
| MEDLINE
| ID: mdl-38647454
ABSTRACT
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Plant Diseases
/
Nicotiana
/
Tobacco Mosaic Virus
/
Ralstonia solanacearum
/
Green Fluorescent Proteins
/
High-Throughput Screening Assays
/
Genetic Vectors
Language:
En
Journal:
Plant J
Journal subject:
BIOLOGIA MOLECULAR
/
BOTANICA
Year:
2024
Document type:
Article
Affiliation country:
China