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
em 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.
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MEDLINE
Assunto principal:
Doenças das Plantas
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Nicotiana
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Vírus do Mosaico do Tabaco
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Ralstonia solanacearum
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Proteínas de Fluorescência Verde
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Ensaios de Triagem em Larga Escala
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Vetores Genéticos
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article